COPYRIGHTpsasir.upm.edu.my/id/eprint/66043/1/FPSK (p) 2016 6 IR.pdf · aktiviti antinosiseptif,...
Transcript of COPYRIGHTpsasir.upm.edu.my/id/eprint/66043/1/FPSK (p) 2016 6 IR.pdf · aktiviti antinosiseptif,...
© COPYRIG
HT UPM
UNIVERSITI PUTRA MALAYSIA
ANTINOCICEPTIVE ACTIVITY, ELUCIDATION OF MECHANISM OF ACTION AND IDENTIFICATION OF BIOACTIVE COMPONENTS OF
Muntingia calabura L. LEAF EXTRACTS
MOHD HIJAZ MOHD SANI
FPSK(p) 2016 6
© COPYRIG
HT UPM
ANTINOCICEPTIVE ACTIVITY, ELUCIDATION OF MECHANISM OF
ACTION AND IDENTIFICATION OF BIOACTIVE COMPONENTS OF
Muntingia calabura L. LEAF EXTRACTS
By
MOHD.HIJAZ MOHD SANI
Thesis submitted to the School of Graduate Studies, University Putra Malaysia, in
Fulfilment of the Requirements for the Degree of Doctor of Philosophy
June 2016
© COPYRIG
HT UPM
COPYRIGHT
All material contained within the thesis, including without limitation text, logos, icons,
photographs and all other artwork, is copyright material of Universiti Putra Malaysia
unless otherwise stated. Use may be made of any material contained within the thesis
for non-commercial purposes from the copyright holder. Commercial use of material
may only be made with the express, prior, written permission of Universiti Putra
Malaysia.
Copyright © Universiti Putra Malaysia
© COPYRIG
HT UPM
i
Abstract of thesis presented to the senate of Universiti Putra Malaysia in fulfilment of
the requirement for the degree of Doctor of Philosophy
ANTINOCICEPTIVE ACTIVITY, ELUCIDATION OF MECHANISM OF
ACTION AND IDENTIFICATION OF
BIOACTIVE COMPONENTS OF MUNTINGIA CALABURA L. LEAF
EXTRACTS
By
MOHD.HIJAZ MOHD SANI
June 2016
Chairman: Associate Professor Zainul Amiruddin Zakaria, PhD
Faculty: Medicine and Health Sciences
Research into plant as an alternative treatment for pain has been widely explored due to
its potentially low adverse effect with great therapeutic activity. Muntingia calabura L.
(Tiliaceae) has recently gained a medicinal status as well as attention from throughout
the world. The present study examined the potential antinociceptive activity of
methanol extract of Muntingia calabura (MEMC) leaves using the acetic acid-induced
abdominal constriction, formalin and hot plate test following the determination of its
safety using acute toxicity test. Then the possible involvement of MEMC-induced
antinociception through capsaicin, glutamate, bradykinin, opioidergic, dopaminergic serotonergic, noradrenergic, adenosisnergic, protein kinase C (PKC), nitric oxide
(NO)/cyclic guanosine monophosphate (cGMP) and potassium channel pathway was
evaluated. Experimental animals (n=6) were pretreated orally with 10% dimethyl
sulfoxide (DMSO; negative control), 5 mg/kg morphine or 100 mg/kg aspirin (positive
control) or 100, 250, and 500 mg/kg of MEMC, followed by the administration of
receptor antagonists and/or induction of nociception. The crude MEMC extract was
further partitioned into three fractions: petroleum ether extract (PEMC), ethyl acetate
extract (EAMC) and aqueous extract (AEMC). The antinociceptive profiling
demonstrates PEMC produced a significantly better activity than EAMC and AEMC.
Possible mechanism of PEMC action was studied using the same assays. Result on the
acute toxicity study shows no mortality and significant behavioural and physiological changes detected. Oral administration of MEMC and PEMC shows a dose-dependent
inhibition in acetic acid-induced abdominal constriction test, formalin-, capsaicin-.
glutamate-, bradykinin- and PMA-induced paw licking test, while only the highest dose
produced significant pain inhibition in hot plate test. Furthermore, the antinociception
caused by MEMC and PEMC in acetic acid-induced abdominal constriction test was
significantly attenuated by intraperitoneal treatment of naloxone (non-specific opioid
receptor antagonist; 5 mg/kg), naltrindole (δ opioid receptor antagonist; 1 mg/kg) nor-
binaltorphimine (κ opioid receptor antagonist; 1 mg/kg), β-funaltraxamine (μ opioid
antagonist; 10 mg/kg), pindolol (5-HT1A receptor antagonist; 1 mg/kg), caffeine (non-
selective adenosine receptor antagonist; 3 mg/kg) and yohimbine (α2 adrenoceptor
antagonist; 0.15 mg/kg). While both extracts did not act on dopaminergic receptor system, PEMC, but not MEMC, was significantly reversed by i.p. injection of atropine
© COPYRIG
HT UPM
ii
(non-selective cholinergic antagonist; 10 mg/kg). At the same time, MEMC and PEMC
were found to inhibit pain through NO/cGMP/PKC as well as potassium channel
pathways. The potential antinociceptive activity seen was probably due to synergistic
effect of flavonoids, tannins, polyphenolic compounds, triterpene and steroid present in
the extracts based on their phytochemical screening. High performance liquid
chromatography analysis shows several peaks, detected at different wavelengths of the
chromatogram, which were suggested to be flavonoid-based compounds. In conclusion,
the synergistic effects of various bioactive components suggested to be responsible in
the antinociceptive activity of MEMC as well as the semi-purified PEMC extract which
involved in central and peripheral pain pathways. The activation of potassium pathway
as well as opioid, adenosinergic, noradrenergic and serotonergic receptors while inhibits NO/cGMP/PKC pathways, TRPV1, glutamate and bradykinin receptors might
be the possible mode of action of both the extracts in exerting their analgesic effect.
© COPYRIG
HT UPM
iii
Abstrak tesis yang dikemukakan kepada senat Universiti Putra Malaysia sebagai
memenuhi keperluan untuk ijazah Doktor Falsafah
AKTIVITI ANTINOSISEPTIF, PENJELASAN MEKANISMA TINDAKAN
DAN MENGENAL PASTI KOMPONEN BIOAKTIF DARI EKSTRAK DAUN
MUNTINGIA CALABURA L.
Oleh
MOHD.HIJAZ MOHD SANI
Jun 2016
Pengerusi: Profesor Madya Zainul Amiruddin Zakaria, PhD
Fakulti: Perubatan dan Sains Kesihatan
Penyelidikan ke atas tumbuhan sebagai perubatan alternatif untuk kesakitan telah luas
diteroka kerana potensi kesan sampingan yang rendah disamping memberi aktiviti
terapeutik yang kuat. Muntingia calabura L. (Tiliaceae) baru-baru ini telah mendapat
status tumbuhan perubatan dan juga perhatian penyelidik dari serata dunia.
Penyelidikan ini mengkaji potensi aktiviti antinosiseptif ekstrak metanol dari daun
Muntingia calabura (MEMC) menggunakan ujian penggeliatan perut mencit, ujian
penjilatan tapak kaki tikus dan ujian plat panas selepas ujian keselamatan toksik akut.
Selepas itu, mekanisma tindakan antinosiseptif MEMC dikaji menggunakan model
eksperimen capsaicin, glutamate, bradykinin, opioidergik, dopaminergic, serotonergic, noradrenergic, adenosinergic, protin kinas C (PKC), saluran nitrik oksida (NO)/kitaran
guanosin monofosfat (cGMP) dan saluran kalium. Haiwan ujikaji (n=6) dirawat secara
oral dengan 10% dimetil sulfoksida (DMSO; kawalan negatif), 5 mg/kg morfin atau
100 mg/kg aspirin (kawalan positif), atau 100, 250 dan 500 mg/kg MEMC, diikuti
dengan pemberian reseptor antagonis dan/atau perangsang kesakitan. Ekstrak kasar
MEMC kemudiannya dibahagikan kepada tiga pecahan: ekstrak petroleum eter
(PEMC), ekstrak etil asetat (EAMC) dan ekstrak akues (AEMC). Mekanisma tindakan
yang mungkin untuk PEMC dikaji menggunakan model eksperimen yang sama.
Keputusan toksik akut menunjukkan tiada kematian serta perubahan signifikan
terhadap tingkah laku dan fisiologi. Pemberian MEMC dan PEMC secara oral
menunjukkan perencatan secara signifikan dalam ujian penggeliatan perut mencit serta ujian penjilatan tapak kaki mencit oleh formalin, capsaicin, glutamate, bradykinin dan
PMA, manakala hanya dos paling tinggi menunjukkan penurunan kesakitan yang
signifikan dalam ujian plat panas. Lebih dari itu, kesan antinosiseptif oleh MEMC dan
PEMC dalam ujian pengeliatan perut mencit dilemahkan secara signifikan selepas
dicabar dengan naloxone (reseptor antagonis opioid tidak spesifik; 5 mg/kg),
naltrindole (reseptor antagonis opioid δ; 1 mg/kg) nor-binaltorphimine (reseptor
antagonis opioid κ; 1 mg/kg), β-funaltraxamine (reseptor antagonis opioid μ; 10
mg/kg), pindolol (reseptor antagonis 5-HT1A; 1 mg/kg), kafein (reseptor antagonis
adenosin tidak spesifik; 3 mg/kg) and yohimbine (reseptor antagonis α2; 0.15 mg/kg).
Walaupun kedua-dua ekstrak tidak bertindak ke atas reseptor dopaminergik, aktiviti
PEMC, tetapi tidak MEMC, secara signifikannya direncat oleh atropin (reseptor antagonis kolinergik tidak spesifik; 10 mg/kg). Pada masa yang sama, MEMC dan
© COPYRIG
HT UPM
iv
PEMC dibuktikan dapat merencat kesakitan melalui saluran NO/cGMP/PKC dan juga
saluran kalium. Potensi aktiviti antinosiseptif yang ditunjukkan berkemungkinan
disebabkan oleh kesan sinergistik flavonoid, tannin, kompaun polyphenolic, triterpene
dan steroid yang ada di dalam ekstrak berdasarkan dari saringan fitokimia yang dibuat.
Analisa kromatografi cecair berprestasi tinggi (HPLC) menunjukkan beberapa puncak,
dikesan dari gelombang berbeza, yang disyorkan sebagai kompaun dari kategori
flavonoid. Kesimpulannya, kesan sinergistik oleh komponen bioaktif yang berbeza
mungkin bertanggungjawab ke atas aktiviti antinosiseptif MEMC dan juga separa-tulen
PEMC yang terlibat dalam saluran kesakitan periferal dan pusat. Pengaktifan saluran
kalium serta reseptor opioid, adenosinergik, noradrenergik and serotonergik disamping
merencat saluran NO/cGMP/PKC serta reseptor TRPV1, glutamate dan bradykinin berkemungkinan ialah mekanisma tindakan yang diambil oleh kedua-dua ekstrak dalam
kesan antinosiseptif yang ditunjukkan.
© COPYRIG
HT UPM
v
ACKNOWLEDGEMENT
Alhamdulillah, all praise and thanks is for Allah subhanahu wa ta’ala for helping me
overcome all the challenges that I had gone through in the study.
I wish to extend my deepest appreciation to my supervisor, Associate Professor Dr.
Zainul Amiruddin Zakaria for his supervision, guidance, valuable advice, patience and
continuous support throughout the course of this project. I truly thank him for giving
me the opportunity to be his postgraduate student. I would like to express my gratitude
and appreciation to my co-supervisors, Associate Professor Dr. Arifah Abdul Kadir and
Dr. Manraj Singh Cheema for their guidance, invaluable advice and support.
Special thanks to my colleague, Tavamani Balan, for her continuous assistance
throughout the study. I would like to thank all my fellow pharmacology lab mates, Kushairi Ahmad, Farhana Yahya, Fauzi Fahmi, Salahuddin and Siti Syariah Mamat for
their companion, cooperation and care towards me. My sincere appreciation dedicates
to the Faculty of Medicine and Heatlth Sciences, Universiti Putra Malaysia for giving
me the opportunity to carry out this project.
Last but not least, my extreme gratitude to my wife, Dr Siti Sarah binti Darussalam for
her patience, support, encouragement and motivation which really help me
accomplished this study. I also owe a depth gratitude to my mother and the family of
my wife for their patience and continuous support.
© COPYRIG
HT UPM
© COPYRIG
HT UPM
vii
This thesis was submitted to the senate of Universiti Putra Malaysia and has been
accepted as fulfilment of the requirement for the degree of Doctor of Philosophy. The
members of the supervisory committee were as follows:
Zainul Amiruddin Zakaria, PhD
Associate Professor
Faculty of Medicine and Health Sciences Universiti Putra Malaysia
(Chairman)
Arifah Abdul Kadir, PhD
Associate Professor
Faculty of Veterinary Medicine
Universiti Putra Malaysia
(Member)
Manraj Singh Cheema, PhD
Senior Lecturer
Faculty of Medicine and Health Sciences Universiti Putra Malaysia
(Member)
_______________________
BUJANG BIN KIM HUAT, PhD
Professor and Dean School of Graduate Studies
Universiti Putra Malaysia
Date:
© COPYRIG
HT UPM
viii
Declaration by graduate student
I hereby confirm that:
This thesis is my original work;
Quotations, illustrations and citations have been duly referenced;
This thesis has not been submitted previously or concurrently for any other degree
at any other institutions;
Intellectual property from the thesis and copyright of thesis are fully-owned by
Universiti Putra Malaysia, as according to the Universiti Putra Malaysia
(Research) Rules 2012;
Written permission must be obtained from supervisor and the office of Deputy Vice-Chancellor (Research and Innovation) before thesis is published (in the form
of written, printed or in electronic form) including books, journals, modules,
proceedings, popular writings, seminar papers, manuscripts, posters, reports,
lecture notes, learning modules, or any other materials as stated in the Universiti
Putra Malaysia (Research) 2012;
There is no plagiarism or data falsification/fabrication in the thesis, and scholarly
integrity is upheld as according to the Universiti Putra Malaysia (Graduate
Studies) Rules 2003 (Revision 2012-2013) and the Universiti Putra Malaysia
(Research) Rules 2012. The thesis has undergone plagiarism detection software.
Signature: _____________________ Date: __________________
Name and Matric No.: Mohd.Hijaz Mohd Sani GS28051
© COPYRIG
HT UPM
ix
Declaration of The Members of Supervisory Committee
This is to confirm that:
The research conducted and the writing of this thesis was under our supervision;
Supervision responsibilities as stated in the Universiti Putra Malaysia (Graduate
Studies) Rules 2003 (revision 2012-2013) are adhered to.
Signature:
Name of Chairman of Supervisory
Committee: Zainul Amiruddin Zakaria, PhD
Signature:
Name of Member of
Supervisory
Committee: Arifah Abdul Kadir, PhD
Signature:
Name of Member of
Supervisory
Committee: Manraj Singh Cheema, PhD
© COPYRIG
HT UPM
x
TABLE OF CONTENTS
Page
ABSTRACT i
ABSTRAK iii
ACKNOWLEDGEMENTS v
APPROVAL vi
DECLARATION viii
LIST OF TABLES xiv
LIST OF FIGURES xv
LIST OF ABBREVATIONS xvii
CHAPTER
1 INTRODUCTION
1.1 Introduction 1
1.2 Hypothesis 2
1.3 Objectives 2
2 LITERATURE REVIEW
2.1 Pain and nociception 3
2.2 Types of pain 4 2.3 Pain theory 6
2.4 Pain processing and sensitization 8
2.5 Pain receptor 9
2.5.1 TRPV1 receptor 10
2.5.2 Glutamate receptor 11
2.5.3 Bradykinin receptor 11
2.5.4 Involvement of protein kinase C 11
2.5.5 Involvement of Nitric Oxide and cGMP pathway 12
2.5.6 Involvement of potassium channels 13
2.5.7 Adenosinergic receptor 13
2.5.8 Serotonergic receptor 14
2.5.9 Cholinergic receptor 14 2.5.10 Dopaminergic receptor 15
2.5.11 Adrenergic receptor 16
2.6 Non-steroidal anti-inflammatory drugs 16
2.7 Opiate and opioid 17
2.7.1 Opioid receptors 18
2.7.2 Endogenous opioid 18
2.7.3 Exogenous opioid 19
2.7.4 Opioid antagonist 20
2.8 Natural product as an alternative analgesic agent 20
2.9 Muntingia calabura L. 21
2.9.1 Traditional uses 22 2.9.2 Scientific findings 24
2.10 Antinociceptive assays 31
2.10.1 Acetic acid-induced abdominal constriction test 31
2.10.2 Hot plate test 32
2.10.3 Formalin test 33
© COPYRIG
HT UPM
xi
3 PREPARATION OF METHANOL EXTRACT OF MUNTINGIA
CALABURA LEAVES (MEMC) AND ITS PARTITIONS: ACUTE
TOXICITY STUDY
3.1 Introduction 34
3.2 Materials and methods
3.2.1 Chemicals 34
3.2.2 Plant collection 34
3.2.3 Methanol extraction process 35
3.2.4 Preparation of petroleum ether, ethyl acetate and
aqueous partition
35
3.2.5 Acute toxicity study 35 3.3 Statistical analysis 36
3.4 Result
3.4.1 Preparation of extracts 36
3.4.2 Acute toxicity study 36
3.5 Discussion 41
4 DETERMINATION OF THE ANTINOCICEPTIVE ACTIVITY
OF MEMC
4.1 Introduction 42
4.2 Materials and methods
4.2.1 Preparation of extract 42 4.2.2 Preparation of drugs 43
4.2.3 Experimental animals 43
4.2.4 Acetic acid-induced abdominal writhing test 43
4.2.5 Hot plate test 43
4.2.6 Formalin test 44
4.3 Statistical analysis 44
4.4 Result
4.4.1 Acetic acid-induced abdominal writhing test 44
4.4.2 Hot plate test 44
4.4.3 Formalin test 44
4.5 Discussion 49
5 POSSIBLE MECHANISM OF ACTION THAT MODULATED
THE ANTINOCICEPTION OF MEMC
5.1 Introduction 52
5.2 Materials and methods
5.2.1 Preparation of extract 52
5.2.2 Preparation of drugs 52
5.2.3 Experimental animals 53
5.2.4 Capsaicin-induced paw licking test 53
5.2.5 Glutamate-induced paw licking test 53
5.2.6 Phorbol 12-myristate 13-acetate (PMA)-induced
nociception
53
5.2.7 Bradykinin-induced nociception 54
5.2.8 Involvement of Nitric Oxide/cGMP pathway 54
5.2.9 Involvement of potassium channels 54
5.2.10 Involvement of various non-opioid receptors 54
5.2.11 Involvement of opioid receptor and its subtype 55
5.3 Statistical analysis 55
© COPYRIG
HT UPM
xii
5.4 Result
5.4.1 Capsaicin-induced paw licking test 55
5.4.2 Glutamate-induced paw licking test 55
5.4.3 Phorbol 12-myristate 13-acetate (PMA)-induced
nociception
55
5.4.4 Bradykinin-induced nociception 56
5.4.5 Involvement of Nitric Oxide/cGMP pathway 56
5.4.6 Involvement of potassium channels 56
5.4.7 Involvement of various non-opioid receptors 56
5.4.8 Involvement of opioid receptor and its subtype 57
5.5 Discussion 67
6 ANTINOCICEPTIVE PROFILE OF VARIOUS PARTITIONS
OBTAINED FROM MEMC: DETERMINATION OF THE MOST
EFFECTIVE PARTITION
6.1 Introduction 73
6.2 Materials and methods 73
6.2.1 Preparation of extract 73
6.2.2 Preparation of drugs 73
6.2.3 Experimental animals 73
6.2.4 Determination of the most effective partition using
acetic acid-induced abdominal constriction test
74
6.2.5 Formalin test 74
6.2.6 Hot plate test 74
6.3 Statistical analysis 74
6.4 Result
6.4.1 Determination of the most effective partition using
acetic acid-induced abdominal constriction test
74
6.4.2 Formalin test 74
6.4.3 Hot plate test 75
6.5 Discussion 79
7 POSSIBLE MECHANISMS OF ACTION THAT MODULATE
ANTINOCICEPTION EXERTED BY PETROLEUM ETHER
PARTITION (PEMC) OF MEMC
7.1 Introduction 81
7.2 Materials and methods
7.2.1 Preparation of extract 81
7.2.2 Preparation of drugs 81
7.2.3 Experimental animals 81
7.2.4 Capsaicin-induced paw licking test 82
7.2.5 Glutamate-induced paw licking test 82
7.2.6 Bradykinin-induced nociception 82
7.2.7 Phorbol 12-myristate 13-acetate (PMA)-induced
nociception
82
7.2.8 Involvement of Nitric Oxide/cGMP pathway 82
7.2.9 Involvement of potassium channels 82
7.2.10 Involvement of various non-opioid receptors 82
7.2.11 Involvement of opioid receptor and its subtype 83
7.3 Statistical analysis 83
7.4 Result
© COPYRIG
HT UPM
xiii
7.4.1 Capsaicin-induced paw licking test 83
7.4.2 Glutamate-induced paw licking test 83
7.4.3 Bradykinin-induced nociception 83
7.4.4 Phorbol 12-myristate 13-acetate (PMA)-induced
nociception
84
7.4.5 Involvement of Nitric Oxide/cGMP pathway 84
7.4.6 Involvement of potassium channels 84
7.4.7 Involvement of various non-opioid receptors 84
7.4.8 Involvement of opioid receptor and its subtype 85
7.5 Discussion 95
8 PHYTOCHEMICAL SCREENING AND HIGH
PERFORMANCE LIQUID CHROMATOGRAPHY ANALYSIS
OF MEMC AND PEMC
8.1 Introduction 97
8.2 Method
8.2.1 Phytochemical screening of dried leaves, methanol
extract and petroleum ether partition of M. calabura
97
8.2.2 HPLC profile of MEMC and PEMC at various
wavelengths
98
8.3 Results 99
8.4 Discussion 106
9 SUMMARY, GENERAL CONCLUSION AND
RECOMMENDATION
107
BIBLIOGRAPHY 111
APPENDICES 135
BIODATA OF STUDENT 137
LIST OF PUBLICATIONS 138
© COPYRIG
HT UPM
xiv
LIST OF TABLES
Table
Page
1 Ethnomedicinal uses of M. calabura
23
2 Pharmacological activities of M. calabura
25
3 Bioactive compounds of M. calabura
26
4 Recorded body weight of control group and rats treated with high dose extract
37
5 Comparison of relative organ weight (in gram) between control
and MEMC-pretreated group obtained at the end of study
38
6 Comparison of complete blood counts between control and
MEMC-pretreated group following the toxicity study
39
7 The comparison of serum biochemistry level between control and
MEMC-pretreated group from the acute toxicity study
40
8 Effect of MEMC on the hot plate test on mice.
47
9 Effect of PEMC in formalin-induced paw licking test.
77
10 Antinociceptive profile of PEMC assessed using the hot-plate test
in mice.
78
11 Comparison on the phytochemical constituents between the leaves
of M. calabura and MEMC.
100
12 Comparison on the phytochemical constituents between the leaves
of M. calabura and PEMC
101
© COPYRIG
HT UPM
xv
LIST OF FIGURES
Figure
Page
1 Descartes' drawing of a pathway-oriented pain mechanism
7
2 Leaves and fruits of Muntingia calabura L.
22
3 Effect of MEMC in acetic acid-induced abdominal
constriction test in mice.
46
4 Effect of MEMC in formalin-induced paw licking test.
48
5 Effect of MEMC on capsaicin-induced paw licking test in rats.
58
6 Effect of MEMC on glutamate-induced paw licking test in
rats.
59
7 Effect of MEMC on PMA-induced paw licking in rats.
60
8 The antinociceptive effect of MEMC against bradykinin-
induced paw licking.
61
9A Effect of L-arginine, L-NAME and their combination on
MEMC antinociception as assessed by acetic acid-induced
abdominal constriction test.
62
9B Effects of L-arginine, methylene blue and their combination
on MEMC antinociception as assessed by acetic acid-induced
abdominal constriction test.
63
10 Effect of MEMC on nociception in the acetic acid-induced
abdominal constriction test in mice following pre-treatment
with potassium channels inhibitors.
64
11 The involvement of several non-opioid receptor antagonists in
MEMC-induced antinociception against acetic acid-induced
abdominal writhing test in mice.
65
12 Analysis of opioid receptor subtypes involvement in MEMC-
induced antinociception against acetic acid-induced writhing
test in mice.
66
13 Comparison between the numbers of abdominal constriction in
mice during the acetic acid-induced abdominal constriction test.
76
14 Effect of PEMC on capsaicin-induced paw licking test in rats.
86
15 Effect of PEMC on glutamate-induced paw licking test in rats.
85
© COPYRIG
HT UPM
xvi
16 The antinociceptive effect of PEMC against bradykinin-
induced paw licking.
93
17 Effect of PEMC on nociception induced by PMA on rats
94
18A Effects of L-arginine, L-NAME and their combination on
PEMC antinociception as assessed by acetic acid-induced
abdominal constriction test.
95
18B Effects of L-arginine, methylene blue and their combination
on PEMC antinociception as assessed by acetic acid-induced abdominal constriction test.
96
19 Effect of PEMC on nociception in the acetic acid-induced
abdominal constriction test in mice following pre-treatment
with potassium channels inhibitors.
97
20 The involvement of several non-opioid receptor antagonists in
PEMC-induced antinociception against acetic acid-induced
abdominal writhing test in mice.
98
21 Analysis of opioid receptor subtypes involvement in PEMC-induced antinociception against acetic acid-induced writhing
test in mice.
99
22 The UV spectra analysis of MEMC
102
23 Chromatogram of MEMC at 254 nm showing the presence of
flavonoids type compounds, namely rutin, quercitrin and
fisetin based on the comparison of their respective UV spectra
analysis
103
24 The HPLC profile of PEMC
104
25 Comparison of the HPLC profile of PEMC against several
pure flavonoids at the wavelength of 300 and 366 nm.
105
26 Graphical abstract summarizing the possible mechanism of
action of MEMC and PEMC.
109
© COPYRIG
HT UPM
xvii
LIST OF ABBREVIATIONS
5-HT Indole 5-hydroxytrptamine/Serotonin
AA Arachidonic acid
ACh Acetylcholine
AEMC Aqueous extract of Muntingia calabura
AMPA α-amino-3-hydroxy-5-methylisoxazole-4-propionic acid
ANOVA Analysis of variance
AR Noradrenergic receptor
ASA Acetylsalicylic acid ATP Adenosine triphosphate
Ca2+ Calcium ion
cAMP Cyclic adenosine monophosphate
cGMP Cyclic guanosine monophosphate
Cl- Chloride ion
CNS Central nervous system
COX Cyclooxygenase
DAG Diacylglycerol
dH2O Distilled water
DMSO Dimethyl sulfoxide
DRG Dorsal root ganglion EAMC Ethyl acetate extract of Muntingia calabura
ED50 Median effective dose
FDA Food and drug association
HPLC High performance liquid chromatography
i.p. Intraperitoneal
i.pl. intraplantar
IASP International association for the study of pain
K+ Potassium ion
L-NAME NG-nitro-L-arginine methyl esters
LOX Lypooxygenase
mACh Muscarinic acetylcholine
MB Methylene blue MEMC Methanol extract of Muntingia calabura
Na+ Sodium ion
nACh Nicotinic acetylcholine
NMDA N-methyl-D-aspartate
NO Nitric oxide
NOS Nitric oxide synthase
NSAIDs Non-steroidal anti-inflammatory drugs
p.o. Orally
PAG periaqueductal grey
PEMC Petroleum ether extract of Muntingia calabura
PGs Prostaglandins PKC Protein kinase C
PLC Phospholipase C
PMA Phorbol 12-myristate 13-acetate
PKA Protein kinase A
PNS Peripheral nervous system
S.E.M. Standard error of mean
© COPYRIG
HT UPM
xviii
sGC Soluble guanylyl cyclase
SNAT Sodium-coupled neural amino acid transporter
TRPV1 Transient receptor potential vanilloid subtype 1
VR1 Vanilloid receptor subtype 1
WHO World health organization
© COPYRIG
HT UPM
1
CHAPTER 1
INTRODUCTION
1.1 Introduction
Pain is a universal experience that can span an enormous spectrum of intensity from
mild discomfort to excruciating agony. In other words, pain is a fundamental and
inevitable form of human suffering, the experience of which is unique to each
individual (Hawthorn and Redmond, 1998). The vocabulary of pain always suggests a
negative event which often relates to an unpleasant experience. Contrary, in basic
biological terms, pain is indeed a protective mechanism. Pain is indispensable for life
and survival that serves a protective function by signalling the presence of noxious,
tissue-damaging condition. From a medical standpoint, the subjective description and
indication of the location of pain may help to pinpoint the underlying cause of disease
(Tortora and Grabowski, 2003).
Human being has been pondering and trying to understand why they feel pain and how
to reduce it. Through the years, understanding and treatment of pain have changed and
developed in accordance with science. Initially viewed religiously as a consequence to
human wrong doing or due to evil spirit entered the body, it has changed to a more
scientific concept. In the first century, the emphasis in medicine shifted from plants to
pills. The usage of natural herbs as analgesia has declined as synthetic drugs with
specific actions were embraced by pharmaceutical industry (Gillis, 1998). However,
these contemporary analgesics, such as opiates and non-steroidal anti-inflammatory
drugs, are often not suitable in all patients and cases, particularly chronic pain on
account of their limitation such as potency, side effects and propensity to lead to
tolerance. As a result, the continuing search for other alternatives is necessary.
Medicinal plants are known to be an important source of new chemical substances with
potential therapeutic effects. The huge discovery of cinchona, morphine, as well as
aspirin which were derived from plant has laid the foundation for further research in
natural plants (Raskin and Ripoll, 2004). The research into plants which are employed
as pain-relievers in traditional ethno-medicine is therefore one of the productive and
logical strategies in the search for new analgesic drugs (Vongtau et al., 2004). Natural
medicine has a net worth of around US$40 billion in global market. The herbal
remedies not only can still be found in ethnic and health food stores, but are also
available in pharmacies and grocery stores (Rasooli, 2011). Data from World Health
Organization (WHO) shows that more than a half of world populations make use of
herbal medicine to relief painful or unpleasant symptoms, with at least 30% are
prescribed (WHO, 1978).
Focusing on natural products as an alternative to many medications, it has been a major
interest not only among scientists but also attract interest from both pharmaceutical
companies as well as financial support from the government. Natural products as
© COPYRIG
HT UPM
2
referred to Holt and Chandra (2002) are herbs, herbal concoctions, dietary
supplements, traditional Chinese medicines or alternative medicines. Natural products
research is guided by ethno-pharmacological knowledge and has brought substantial
contributions to drug innovation by providing novel chemical structures and/or
mechanism of actions (Rates, 2001).
1.2 Hypothesis
MEMC possess significant antinociceptive activity due to the presence of
flavonoids-based compounds.
1.3 Objectives
General:
a) To evaluate the antinociceptive activity of crude and the most effective
partition of methanol extract of Muntingia calabura (MEMC).
Specific:
a) To determine the safety of MEMC using single high-dose acute toxicity model
b) To determine antinociceptive profiles of MEMC using various animal models
c) To elucidate the possible mechanisms of action that take part in the
antinociception of MEMC
d) To determine the most effective partition of MEMC using acetic acid-induced
abdominal constriction test.
e) To elucidate the possible mechanisms of action that take part in the
antinociception of PEMC
f) To elucidate and identify the possible bioactive compounds responsible for the
MEMC and PEMC triggered antinociception using the phytochemical
screening test and HPLC analysis
© COPYRIG
HT UPM
111
BIBLIOGRAPHY
Aanonsen, L.M. and Wilcox, G.L. 1987. Nociceptive action of excitatory amino acids
in the mouse: effects of spinally administered opioids, phencyclidine and sigma
agonists, Journal of Pharmacology and Experimental Therapeutics. 243: 9-19.
Aanonsen, L.M. and Wilcox, G.L. 1990. Excitatory amino acid receptors and
nociceptive neurotransmission in rat spinal cord, Pain. 41: 309-321.
Abacioglu, N., Tun Tan, B., Akbulut, E. and Akici, I. 2000. Participation of the
components of L-arginine/nitric oxide/cGMP cascade by chemically-induced
abdominal constriction in the mouse, Life Science. 67: 1127-1137.
Abbott, F.V., Franklin, K.B.J. and Westbrook, R.F. 1995. The formalin test: scoring
properties of the first and second phases of the pain response in rats, Pain. 60:
91-102.
Aley, K., Messing, R., Mochly-Rosen, D. and Levine, J. 2000. Chronic hypersensitivity
for inflammatory nociceptor sensitization mediated by the epsilon isozyme of
protein kinase C, Journal of Neuroscience. 20: 4680–4685.
Aley, K.O., McCarter, G. and Levine, J.D. 1998. Nitric oxide signaling in pain and
nociceptor sensitization in the rat, Journal of Neuroscience. 18: 7008–7014.
Alkondon, M. and Albuquerque, E.X. 1993. Diversity of nicotinic acetylcholine
receptors in rat hippocampal neurons: I. Pharmacological and functional
evidence for distinct structural subtypes, Journal of Pharmacology and
Experimental Therapeutics. 265: 1455–1473.
Altier, N. and Stewart, J. 1998. Dopamine receptor antagonists in the nucleus
accumbens attenuate analgesia induced by ventral tegmental area substance P or
morphine and by nucleus accumbens amphetamine, Journal of Pharmacology
and Experimental Therapeutics. 285: 208–215.
Altier, N., and Stewart, J. 1999. The role of dopamine in the nucleus accumbens in
analgesia, Life Science. 65: 2269–2287.
Amenta, F., Ferrante, F. and Ricci, A. 1995. Pharmacological characterization and
autoradiographic localization of dopamine receptor subtypes in the
cardiovascular system and in the kidney. Hypertension research: official journal
of the Japanese society of hypertension. 18: s23-27.
Ankier, S.I. 1974. New hot plate tests to quantify antinociceptive and narcotic
antagonist activities, European Journal of Pharmacoogyl. 27: 1-4.
Archer, S.L., Huang, J.M., Hamp, l.V., Nelson, D.P., Shultz, P.J., and Weir, E.K.,
1994. Nitric oxide and cGMP cause vasorelaxation by activation of a
charybdotoxin-sensitive K+ channel by cGMP-dependent protein kinase,
Proceeding of the National Academy of Science, USA. 91: 7583–7587.
© COPYRIG
HT UPM
112
Aznam, N. Atun, S. Arianingrum, R. and Nurestri, S. 2012. Isolation, identification and
antiviral activity of bioactive compounds of Kampheria rotunda, International
Proceedings of Chemical, Biological & Environmental Engineering. 38: 27-30.
Balan, T, Mohd Sani, M.H., Suppaiah, V., Mokhtaruddin, N., Suhaili, Z. Ahmad, Z et
al. 2013. Antiulcer activity of Muntingia calabura leaves involves the
modulation of endogenous nitric oxide and nonprotein sulfhydryl compounds,
Pharmaceutical Biology. 52: 410–18.
Bandeira, G.N., da Camara, C.A.G., de Moraes, M.M., Barros, R., Muhammad, S. and
Akhtar, Y. 2013. Insecticidal activity of M. calabura extracts against larvae and
pupae of diamondback, Plutella xylostella (Lepidoptera, Plutellidae). Journal of
King Saud University – Science. 25: 83–89.
Bantel, C., Maze, M., Stone, L., and Wilcox, G. α2-adrenergic agonists in pain
treatment in Encyclopedia of pain ed. Schmidt, R.F. and Willis, W.D. New
York: Springer-verlag Berlin Heilberg, 2007.
Basbaum, A.I. and Bushnell, M.C. Science of Pain. San Diego: Academic Press, 2009.
Beirith, A., Santos, A.R., Calixto, J.B. 2002. Mechanisms underlying the nociception
and paw oedema caused by injection of glutamate into the mouse paw, Brain
Research. 924: 219-228.
Beirith, A., Santos, A.R.S., Calixto, J.B., Hess, S.C., Messana, I., Ferrari, F. and Yunes,
R.A. 1999. Study of the antinociceptive action of the ethanolic extract and the
triterpene 24-hydroxytormentic acid isolated from the stem bark of Ocotea
suaveolens, Planta Medica. 65: 50–55.
Beitz, A.J. Descending circuitry, transmitters and receptors in Encyclopedia of pain ed.
Schmidt, R.F. and Willis, W.D. New York: Springer-verlag Berlin Heilberg.
2007.
Belmonte, C. and Cervero, F. Neurobiology of Nociceptors: Oxford University Press,
1996.
Belmonte, C. and Viana, F. Transduction and encoding of noxious stimuli in
Encyclopedia of pain ed. Schmidt, R.F. and Willis, W.D. New York: Springer-
verlag Berlin Heilberg. 2007.
Bentley, G.A., Newton, S.H. and Starr, J. 1981. Evidence for an action of morphine
and the enkephalines on sensory nerve endings in the mouse peritoneum, British
Journal of Pharmacology. 73: 325-332.
Bentley, G.A., Newton, S.H. and Starr, J. 1983. Studies on the antinociceptive action of
a-agonist drugs and their interactions with opioid mechanisms, British Journal
of Pharmacology. 79: 125–134.
Berne, M.R. and Levy, N.M. Principles of Physiology. Mosby International. 2000.
© COPYRIG
HT UPM
113
Berrazueta, JR., Losada, A., Poveda, J., Ochoteco, A., Riestra, A., Salas, E. et al. 1996.
Successful treatment of shoulder pain syndrome due to supraspinal tendinitis
with transdermal nitroglycerin. A double blind study. Pain. 66: 63-67.
Besson, J.M. 1999. The neurobiology of pain, The Lancet. 353: 1610-1615.
Bhave, G. Karim, F. Carlton, S. M. and Gereau, R.W. 2001. Peripheral group I
metabotropic glutamate receptors modulate nociception in mice, Nature
Neuroscience. 4: 417-423.
Bian, K., Ke, Y., Kamisaki, Y. and Murad, F. 2006. Proteomic modification of nitric
oxide, Journal of Pharmacological Sciences. 101: 271-279.
Bolotina, V.M., Najibi, S., Palacino, J.J., Pagano, P.J. and Cohen, R.A. 1994. Nitric
oxide directly activates calcium-dependent potassium channels in vascular
smooth muscle, Nature. 368: 850–853
Bredt, D.S. and Snyder, S.H. 1992. Nitric oxide, a novel neuronal messenger, Neuron.
8: 3–11.
Brown, D.A. 2010. Muscarinic acetylcholine receptors (mAChRs) in the nervous
system: some functions and mechanisms, Journal of Molecular Neuroscience,
41: 340-346.
Brunori, M. Giuffre, A. Sarti, P., Stubauer, G. and Wilson, M.T. 1999. Nitric oxide and
cellular respiration, Cellular and Molecular Life Science. 56: 549–557.
Burian, M. and Geisslinger, G. 2005. COX-dependent mechanisms involved in the
antinociceptive action of NSAIDs at central and peripheral sites, Pharmacology
and therapeutic. 107: 139-154.
Burnstock, G. 2008. Purinergic signalling and disorders of the central nervous system,
Nature Reviews Drug Discovery. 7: 575–90.
Cadiou, H., Studer, M., Jones, N.G., Smith, E.S.J., Ballanrd, A., McMohan, S.B., et al.
2007. Modulation of acid-sensing ion channel activity by nitric oxide, Journal
of Neuroscience. 27: 132-151.
Cahill, C.M., Morinville, A., Lee, M.C., Vincent, J.P., Collier, B. and Beaudet, A.
2001. Prolonged morphine treatment targets delta opioid receptors to neuronal
plasma membranes and enhances delta-mediated antinociception, Journal of
Neuroscience. 21: 7598-7607.
Calixto, J.B., Beirith, A., Ferreira, J., Santos, A.R.S., Filho, V.C. and Yunes, R.A.
2000. Naturally occurring antinociceptive substances from plants, Phytotherapy
research. 14: 401-418.
Calixto, J.B., Cabrini, D.A., Ferreira, J. and Campos, M.M. 2000a. Kinins in pain and
inflammation. Pain. 87: 1–5.
© COPYRIG
HT UPM
114
Calixto JB, Santos ARS, Cechinel Filho V, Yunes RA. 1998. A review of the plants of
the genus Phyllanthus: their chemistry, pharmacology, and therapeutic potential,
Medicinal Research Review. 18: 225-258.
Carpenter, S.E. and Lynn, B. 1991. Vascular and sensory responses of human skin to
mild injury after topical treatment with capsaicin, British Journal of
Pharmacology. 73: 755-758.
Carrier, G.O., Fuchs, L.C., Winecoff, A.P., Giulumian, A.D. and White, R.E. 1997.
Nitro-vasodilators relax mesenteric microvessels by cGMP-induced stimulation
of Ca2+
-activated K+ channels, American Journal of Physiology. 273: H76–H83.
Carrol, I., Mackey, S. and Gaeta, R. 2007. The role of adrenergic receptors and pain:
The good, the bad, and the unknown, Seminars in Anesthesia, Perioperative
Medicine and Pain. 26: 17-21.
Carroll, D. and Bowsher, D. Pain Management and Nursing Care. Oxford:
Butterworth-Heinemann Ltd, 1995.
Carvalho, J.C., Silva, M.F., Maciel, M.A. et al. 1996. Investigation of anti-
infammatory and antinociceptive activities of transdehydrocrotonin, a 19-nor-
clerodane diterpene from Croton cajucara. Part 1, Planta Medica. 62: 402-404.
Caterina, M. and Julius, D. 2001. The vanilloid receptor: a molecular gateway to the
pain pathway, Annual Review of Neuroscience. 24: 487-517.
Caterina, M.J., Schumacher, M.A., Tominaga, M., Rosen, T.A., Levine, J.D. and
Julius, D. 1997. The capsaicin receptor: a heat-activated ion channel in the pain
pathway, Nature. 389: 816-824.
Caulfield, M.P. and Birdsall, N.J. 1998. International Union of Pharmacology. XVII.
Classification of muscarinic acetylcholine receptors, Pharmacological Reviews.
50: 279-290.
Cervero, F. 2009a. Pain: Friend or Foe? A Neurobiologic Perspective: The 2008
Bonica Award Lecture, Regional Anesthesia and Pain Medicine. 34: 569-574.
Cervero, F. and Laird, J.M.A. 1991. One pain or many pains? A new look at pain
mechanisms, New Physiology Science. 6: 268-273.
Cervero, F. and Laird, J.M.A. 1996a. Mechanism of touch-evoked pain (allodynia): a
new model, Pain. 68: 13-23
.
Cervero, F. Pain theories, in Science of Pain ed. Basbaum, A.I and Bushnell, M.C. San
Diego: Academic Press. 2009.
Cervero, F., Kruger, L., Kumazawa, T., and Kazue, M. 1996. Spinal cord mechanisms
of hyperalgesia and allodynia: role of peripheral input from nociceptors,
Progress in Brain Research. 113: 413-422.
© COPYRIG
HT UPM
115
Cesare, P., Dekker, L.V., Sardini, A., Parker, P.J. and McNaughton, P.A. 1999.
Specific involvement of PKCε in sensitization of the neuronal response to
painful heat, Neuron. 23: 617–624.
Chan, K., Islam, M.W., Kamil, M., Radhakrishna, R., Zakaria, M.N.M., Habibullah, M.
and Attas, A. 2000. The analgesic and anti-inflammatory effects of
Portulacaoleracea L. subsp. Sativa (Haw) Celak, Journal of
Ethnopharmacology. 73: 445-451.
Chan, T.F., Tsai, H.Y. and Tian-Shang, W. 1995. Anti-inflammatory and analgesic
activities from the roots of Angelica pubescens, Planta Medica. 61: 2–8.
Chandrasekharan, N.V., Dai, H., Roos, K.L., Evanson, N.K., Tomsik, J., Elton, T.S., et
al. 2002. COX-3, a cyclooxygenase-1 variant inhibited by acetaminophen and
other analgesic/antipyretic drugs: cloning, structure, and expression,
Proceedings of the National Academy of Sciences. USA. 99: 13926–13931.
Chen, J.J., Lee, H.H., Shih, C.D., Liao, C.H., Chen, I.S. and Chou, T.H. 2007. New
dihydrochalcones and anti-platelet aggregation constituents from the leaves of
Muntingia calabura, Planta Medica. 73: 572–577.
Chen, J.J., Lin, R.W., Duh, C.Y. and Huang, H.Y. 2004. Flavones and cytotoxic
constituents from the stem bark of Muntingia calabura, Journal-Chinese
Chemical Society Taipei. 51: 665–670.
Chen, S.R., Wess, J. and Pan, H.L. 2005. Functional activity of the M2 and M4 receptor
subtypes in the spinal cord studied with muscarinic acetylcholine receptor
knockout mice, Journal of Pharmacology and Experimental Therapeutics. 313:
765-770.
Choi, J.H. Hung, B.H. Kang, O.H. Choi, H.J. Park, P.S. Cho, S.H. et al. 2006. The anti-
inflammatory and anti-nociceptive effects of ethyl acetate fraction of Cynanchi
paniculati Radix, Biological Pharmacology Bulletin. 29: 971–975.
Chuang, H.H., Prescott, E.D., Kong, H., Shields, S., Jordt, S.E., Basbaum, A.I. et al.,
2001. Bradykinin and nerve growth factor release the capsaicin receptor from
PtdIns(4,5)P2-mediated inhibition, Nature. 411: 957–962.
Chung, J.M. Animal models and experimental tests to study nociception and pain, in
Encyclopedia of Pain ed. Schmidt, R.F. and Willis, W.D. New York: Springer-
Verlag Berlin Heidelberg. 2007.
Clark, J.D. and Tempel, B.L. 1998. Hyperalgesia in mice lacking the Kv1.1 potassium
channles gene, Neuroscience letter. 251: 121-124.
Coderre, T.J. 1993. The role of excitatory amino acid receptors and intracellular
messengers in persistent nociception after tissue injury in rats, Molecular
Neurobiology. 7: 229-246.
Coderre, T.J. Noxious stimulus-induced plasticity in spinal cord dorsal horn: evidence
and insights on mechanisms obtained using the formalin test in Spinal cord
© COPYRIG
HT UPM
116
plasticity: alteration in reflex function ed. Patterson, M.M. and Grau, J.W.
Boston: Kluwer academic publisher. 2001.
Coderre, T.J. Spinal cord mechanisms of hyperalgesia and allodynia in Science of Pain
ed. Basbaum, A.I. and Bushnell, M.C. San Diego: Academic Press. 2009.
Coderre, T.J., Fundytus, M.E., McKenna, J.E., Dalal, S. and Melzack, R. 1993. The
formalin test: A validation of the weighed-scored method of behavioral pain
rating, Pain. 54: 43-50.
Coggeshall, R.E. and Carlton, S.M. 1997. Receptor localization in the mammalian
dorsal horn and primary afferent neurons, Brain Research Review. 24: 28-66.
Collier, H.O.J, Dinneen, J.C., Johnson, C.A. and Schneider, C. 1968. The abdominal
constriction response and its suppression by analgesic drugs in the mouse,
British Journal of Pharmacology and Chemotherapy. 32: 295–310.
Cui, M. Honore, P. Zhong, C. Gauvin, D. Mikusa, J. Hernandez, G., et al. 2006.
TRPV1 receptors in the CNS play a key role in broad-spectrum analgesia of
TRPV1 antagonists, Journal of Neuroscience. 26: 9385–9393.
Cunha, T.M., Roman-Campos, D., Lotufo, C.M., Duarte, H.L., Souza, G.R., Verri Jr.,
W.A., et al. 2010. Morphine peripheral analgesia depends on activation of the
PI3Kγ/AKT/nNOS/NO/KATP signaling pathway, Proceedings of the National
Academy of Sciences USA. 10: 4442-4447.
da Silva, R.S. 2011. Caffeine, Reproductive and Developmental toxicology. 27: 355-
364.
Dahlström, A. and Fuxe, K. 1964. Evidence for the existence of monoamine-containing
neurons in the central nervous system, Acta Physiologica Scandinavica. 62: 3-
55.
Damas, J, Bourdon, V, Remacle-Volon, G, Lecomte, J. 1985. Proinflammatory
flavonoids which are inhibitors of prostaglandin biosynthesis, Prostaglandins
Leukotrienes and Medicine. 19: 11-24.
Davies, S.N. and Lodge, D. 1987. Evidence for involvement of N-methylaspartate
receptors in 'wind-up' of class 2 neurons in the dorsal horn of the rat, Brain
Research. 424: 402-406.
Dawson, T.M. and Snyder, S.H. 1994. Gases as biological messengers: nitric oxide and
carbon monoxide in the brain, Journal of Neuroscience. 14: 5147–5159.
De Moura, R.S., Rios, A.A.S., Santos, E.J.A., Nascimento, A.B.A., Resende, A.C.,
Neto, M.L. et al. 2004. Role of NO-cGMP pathway in the systemic
antinociceptive effect of clonidine in rats and mice, Pharmacology,
biochemistry and behavior. 78: 247-253.
© COPYRIG
HT UPM
117
De Smet, P.A.G.M. 1997. The role of plant-derived drugs and herbal medicines in
healthcare, Drugs. 54: 801-840.
De Souza, M.M., Pereira, M.A., Adenghi, A.V., Mora, T.C., Bresciani, L.F., Yunes,
R.A. et al. 2009. Filicene obtained from Adiantum cuneatum interacts with the
cholinergic, dopaminergic, glutamatergic, GABAergic and tachykinergic
systems to exert antinociceptive effect in mice, Pharmacology biochemistry and
behavior. 93: 40-46.
Denninger, J.W. and Marletta, M.A. 1999. Guanylatecyclase and the NO/cGMP
signaling pathway, Biochimica et Biophysica Acta. 1411: 334-350.
Deraedt, R., Jouquey, S., Delevallee, F. and Flahaut, M. 1980. Release of
prostaglandins E and F in an algogenic reaction and its inhibition, European
Journal of Pharmacology. 61: 17-24.
Descartes, R. 1664. L'Homme. Chez Jacques Le Gras.
Dickenson, A.H. and Sullivan, A.F. 1987. Evidence for a role of the NMDA receptor in
the frequency dependent potentiation of deep rat dorsal horn nociceptive
neurones following C fiber stimulation, Neuropharmacology. 26: 1235-1238.
Diguangco, J. Notes on Philippine Medicinal Plants, Manila, Philippines, 1959.
Dray, A. 1992. Mechanism of action of capsaicin-like molecules on sensory neurones,
Life Sciences. 51: 1759-1765.
Dray, A. and Perkins, M. Kinins and pain in The handbook of immunopharmacology:
the kinin system ed. Farmer, S.G. London: Academic Press, 1997.
Dray, A. Pharmacological modulation of pain in Science of Pain ed. Basbaum, A.I. and
Bushnell, M.C. San Diego: Academic Press. 2009.
Duarte, I.D., Lorenzetti, B.B and Ferreira, S.H. 1990. Peripheral analgesia and
activation of the nitric oxide-cyclic GMP pathway, European Journal of
Pharmacology. 186: 289–293.
Dubuisson, D. and Dennis, S.G. 1977. The formalin test: a quantitative study of the
analgesic effects of morphine, meperidine and brain stem stimulation in the rats
and cats, Pain. 4: 161-174.
Duttaroy, A., Gomeza, J., Gan, J.W., Siddiqui, N., Basile, A.S., Harman, W.D. et al.
2002. Evaluation of muscarinic agonist-induced analgesia in muscarinic
acetylcholine receptor knockout mice, Molecular pharmacology. 62: 1084-
1093.
Eisenach, J.C., Detweiler, D.J., Tong, C., D’Angelo, R., and Hood, D.D. 1996.
Cerebrospinal fluid norepinephrine and acetylcholine concentrations during
acute pain, Anesthiology and Analgesia. 82: 621-626.
© COPYRIG
HT UPM
118
Espejo, E.F. and Gil, E. 1998. Antagonism of peripheral 5-HT4 receptors reduces
visceral and cutaneous pain in mice, and induces visceral analgesia after
simultaneous inactivation of 5-HT3 receptors, Brain Research. 788: 20–24.
Farnsworth, N.R. and Bingel, A.S. 1977. Problems and prospects of discovering new
drugs from higher plants by pharmacological screening, Preceedings in Life
Sciences. 1-22.
Ferreira, J., da Silva, G.L., Calixto, J.B. 2004. Contribution of vanilloid receptors to the
overt nociception induced by B2 kinin receptor activation in mice, British
Journal of Pharmacology. 141: 787-794.
Ferreira, J., Triches, K.M., Medeiros, R., Calixto, J.B. 2005. Mechanisms involved in
the nociception produced by peripheral protein kinase C activation in mice,
Pain. 117: 171–181.
Ferreira, S. 1972. Prostaglandins, aspirin-like drugs and analgesia, Nature: New
Biology. 240: 200–203.
Fidecka, S. and Lalewicz, S. 1997. Studies on the antinociceptive effects of sodium
nitroprusside and molsidomine in mice, Polish Journal of Pharmacology. 49:
395–400.
Forstermann, U. and Sessa, W.C. 2012. Nitric oxide synthases: regulation and function,
European Heart Journal. 33: 829-837.
Fredholm, B.B., Battig, K., Holmen, J., Nehlig, A. and Avartau, E.E. 1999. Actions of
caffeine in the brain with special reference to factors that contribute to its
widespread use, Pharmacological Reviews. 51: 83–133.
Fredholm, B.B., Chen, J.F., Cunha, R.A., Svenningsson, P. and Vaugeois, J.M. 2005.
Adenosine and brain function, International Review of Neurobiology. 63: 191-
270.
Fredholm, B.B., Ijzerman, A.P., Jacobson, K.A., Klotz, K.N., and Linden, J. 2001.
International union of pharmacology XXV. Nomenclature and classification of
adenosine receptors, Pharmacological Reviews. 53: 527–552.
Fu, J.Y., Masferrer, J.L., Seibert, K., Raz, A. and Needleman, P. 1990. The induction
and suppression of prostaglandin H2 synthase (cyclooxygenase) in human
monocytes, Journal of Biological Chemistry. 265: 16737–16740.
Fundytus, M.E. 2001. Glutamate receptors and nociception: implications for the drug
treatment of pain, CNS Drugs. 15: 29–58.
Galvez, J., Cruz, T., Crespo, E., Ocete, M.A., Lorente, M.D., Sánchez de Medina, F.
and Zarzuelo, A. 1997. Rutoside as mucosal protective in acetic acid-induced rat
colitis, Planta Medica. 63: 409–414.
Garcia-Martinez, C., Humet, M., Planells-Casas, R., Gomis, A., Capri, M., Viana, F., et
al. 2002. Attenuation of thermal nociception and hyperalgesia by VR1 blockers.
Proceeding of the National Academy of Science, U.S.A. 99: 2374-2379.
© COPYRIG
HT UPM
119
Garthwaite, J. and Boulton, C.L. 1995. Nitric oxide signaling in the central nervous
system, Annual Review of Physiology. 57: 683–706.
Geisslinger, G. Non-steroidal anti-inflammatory drugs (NSAIDs) in Encyclopedia of
pain ed. Schmidt, R.F. and Willis, W.D. 2007. New York: Springer-verlag
Berlin Heilberg. 2007.
Giglio, C.A., Defino, H.L.A., da Silva, C.A., de-Souza, A.S. and del Bel, E.A. 2006.
Behavioral and physiological methods for early quantitative assessment of
spinal cord injury and prognosis in rats, Brazil Journal of Medical Biology
Research, 39: 1613–1623.
Gilchrist, H.D., Allard, B.L. and Simone, D.A. 1996. Enhanced withdrawal responses
to heat and mechanical stimuli following interplantar injection of capsaicin in
rats, Pain. 67: 179-188.
Glazer, E.J., Steinbusch, H., Verhofstad, A., and Basbaum, A.I. 1981. Serotonin
neurons in nucleus raphe dorsalis and paragigantocellularis of the cat contain
enkephalin, Journal of Physiology (Paris). 77: 241-245.
Goncalves, C.E., Araldi, D., Panatieri, R.B., Rocha, J.B., Zeni, G., and Nogueira, C.W.
2005. Antinociceptive properties of acetylenic thiophene and furan derivatives:
evidence for the mechanism of action, Life Sciences. 76: 2221–2234.
Gordh, T., Sharma, H.S., Alm, P. and Westman, J. 1998. Spinal nerve lesion induces
upregulation of neuronal nitric oxide synthase in the spinal cord. An
immunohistochemical investigation in the rat, Amino Acids. 14: 105–112.
Gybels, J.M. and Sweet W.H. Neurosurgical treatment of persistent pain in Pain and
Headache ed. Gildenberg, Ph.L. Basel: Karger. 1989.
Hargreaves, K., Dubner, R., Brown, F. and Joris, J. 1988. A new and sensitive method
for measuring thermal nociception in cutaneous hyperalgesia, Pain. 32: 77-88.
Havsteen, B.H. 2002. The biochemistry and medical significance of flavonoids,
Pharmacology and Therapeutics. 96: 67-202.
Hawthorn, J. and Redmond, K. Pain: Causes and Management. Oxford: Blackwell
Science Ltd, 1998.
Heapy, C.G., Jamieson, A. and Russell N.J.W. 1987. Afferent C-fiber and A-delta
activity in models of inflammation, British Journal Pharmacology. 90: 164.
Higgs, J., Wasowski, C., Loscalzo, L.M. and Marder, M. 2013. In vitro binding
affinities of a series of flavonoids for μ-opioid receptors. Antinociceptive effect
of the synthetic flavonoid 3,3-dibromoflavanone in mice, Neuropharmacology.
72: 9-19.
Hebbes, C. and Lambert, D. 2007. Non-opioid analgesic drugs. Anasthesia and
intensive care medicine. 9: 79-83.
© COPYRIG
HT UPM
120
Höglund, A.U. and Baghdoyan, H.A. 1997. M2, M3 and M4, but not M1, muscarinic
receptor subtypes are present in rat spinal cord. Journal of Pharmacology and
Experimental Therapeutics. 281: 470-477.
Holzmann, A. Manktelow, C., Weimann, J., Bloch, K.D. and Zapol, W.M. 2001.
Inhibition of lung phosphodiesterase improves responsiveness to inhaled nitric
oxide in isolated-perfused lungs from rats challenged with endotoxin, Intensive
Care Medicine. 27: 251–257.
Honda, K., Harada, A., Takano, Y. and Kamiya, H. 2000. Involvement of M3
muscarinic receptors of the spinal cord in formalin-induced nociception in mice,
Brain Research. 859: 38-44.
Hosseinzadeh, H. and Younesi, H.M. 2002. Antinociceptive and anti-inflammatory
effects of Crocus sativus L. stigma and petal extracts in mice, BMC
Pharmacology. 2: 7.
Huang, S.M., Bisogno, T., Trevisani, M., Al-Hayani, A., de Petrocellis, L., Fezza, F., et
al. 2002. An endogenous capsaicin-like substance with high potency at
recombinant and native vanilloid VR1 receptors, Proceedings of the National
Academy of Science, U.S.A. 99: 8400–8405.
Hughes, J., Smith, T.W., Kosterlitz, H.W., Fotherqill, L.A., Morgan, B.A., and Morris,
H.R. 1975. Identification of two related pentapeptides from the brain with the
potent opiate agonist activity. Nature (Lond). 58: 577-579.
Hugonin, L., Vukojević, V., Bakalkin, G. and Gräslund, A. 2006. Membrane leakage
induced by dynorphins, FEBS Letters. 580: 3201-3205.
Hunskaar, H.S., Fasmer, O.B. and Hole, K. 1985. Formalin test in mice, a useful
technique for evaluating mild analgesics, Journal of Neuroscience Methods. 14:
69-76.
Hunt, S.P. and Mantyh, P.W. 2001. The molecular dynamics of pain control. Nature
Reviews Neuroscience. 2: 83-91.
Hurley, R.W. and Hammond, D.L. 2001. Contribution of endogenous enkephalins to
the enhanced analgesic effects of supraspinal {micro} opioid receptor agonists
after inflammatory injury, Journal of Neuroscience. 21: 25-36.
Iadarola, M.J., Brady, L.S., Draisci, G., and Dubner, R. 1988. Enhancement of
dynorphin gene expression in spinal cord following experimental inflammation:
stimulus specificity, behavioral parameters and opioid receptor binding,
Pain. 35: 313-326.
Ibrahim, I.A.A., Abdulla, M.A., Abdelwahab, S., Al-Bayaty, F. and Abdul Majid, N.
2012. Leaves extract of Muntingia calabura protects against gastric ulcer
induced by ethanol in Sprague–Dawley rats, Clinical and Experimental
Pharmacology. DOI:10.4172/2161-1459.S5-004.
© COPYRIG
HT UPM
121
Ikeda, H., Stark, J., Fischer, H., Wagner, M., Drdla, R., Jager T. et al. 2006. Synaptic
amplifier of inflammatory pain in the spinal dorsal horn. Science. 312: 1659-
1662.
Ikeda, Y., Ueno, A., Narada, H. and Oh-ishi, S. 2001. Involvement of vanilloid
receptor VR1 and prostanoids in the acid-induced writhing responses on mice,
Life sciences. 69: 2911-2919.
Ing, D., Zang, J., Dzau, V., Webster, K. and Bisphopric, N. 1999. Modulation of
cytokine-induced cardiac myocyte apoptosis by nitric oxide Bak- and Bal-x,
Circulation Research. 84: 21-33.
Irving, G.A. and Wallance, M.S. Pain Management for the Practicing Physician. New
York: Churchill Livingstone Inc. 1996.
Ito, A., Kumamoto, E., Takeda, M., Shibata, K., Sagai, H. and Yoshimura, M. 2000.
Mechanisms for ovariectomy-induced hyperalgesia and its relief by calcitonin:
participation of 5- HT1A-like receptor on C-afferent terminals in substantia
gelatinosa of the rat spinal cord, Journal of Neuroscience. 20: 6302–6308.
Iwamoto, E.T. and Marion, L. 1993. Characterization of the antinociception produced
by intrathecally administered muscarinic agonists in rats, Journal of
Pharmacology and Experimental Therapeutics. 266: 329-338.
Jajoo, S., Mukherjea, D., Watabe, K. and Ramkumar, V. 2009. Adenosine A3 receptor
suppresses prostate cancer metastasis by inhibiting NADPH oxidase activity,
Neoplasia. 11: 1132–1145.
Janig, W. 2009. Autonomic nervous system and pain in Science of Pain ed. Basbaum,
A.I. and Bushnell, M.C. San Diego: Elsevier Inc. 2009.
Jhaveri, M.D., Elmes, S.J., Kendall, D.A. and Chapman, V. 2005. Inhibition of
peripheral vanilloid TRPV1 receptors reduces noxious heat-evoked responses of
dorsal horn neurons in naïve, carrageenan-inflamed and neuropathic
rats, European Journal of Neuroscience. 22: 361–370.
Ji, R.R. and Woolf, C.J. 2001. Neuronal plasticity and signal transduction in
nociceptive neurons: implications for the initiation and maintenance of
pathological pain, Neurobiology of Disease. 8: 1–10.
Jones, P.G and Dunlop, J. 2007. Targeting the cholinergic system as a therapeutic
strategy for the treatment of pain, Neuropharmacology, 53: 197-206.
Julie, G.H. Serotonin in Basic neurochemistry: molecular, cellular and medical aspects
ed. Siegel, G.J., Albers, R.W., Brady, S.T. and Price, D.L. San Diego: Elsevier
academic press. 2006.
Julius, D. and Basbaum, A.I. 2001. Molecular mechanism of nociception, Nature. 413:
203-210.
© COPYRIG
HT UPM
122
Julius, D. and Basbaum, A.I. 2001. Molecular mechanisms of nociception, Nature
(London). 413: 203–210.
Kandel, E.R., Schwartz, J.H. and Jessell, T.M. Principles of Neural Science. New York:
McGraw-Hill. 2000.
Kaneda, N., Pezzuto, J.M., Soejarto, D.D., Kinghorn, A.D., Farnworth, N.R., Santisuk,
T. 1991. Plant anticancer agents, XLVIII. New cytotoxic flavonoids from
Muntingia calabura roots, Journal of Natural Product. 54: 196–206.
Karlsten, R., Gordh, T. and Post, C. 1992. Local antinociceptive and hyperalgesic
effects in the formalin test after peripheral administration of adenosine
analogues in mice, Pharmacology and Toxicology. 70: 434-438.
Karumi, Y., Onyeyili, P. and Ogugbuaja, V.O. 2003. Anti-inflammatory and
antinociceptive (analgesic) properties of Momordical balsamina Linn. (Balsam
apple) leaves in rats, Pakistan Journal of Biological Science. 6: 1515–1518.
Katavic, P.L., Lamb, K., Navarro, H. and Prisinzano, T.E. 2007. Flavonoids as opioid
receptor ligands: identification and preliminary structure-activity relationships,
Journal of Natural Product. 70: 1278-1282.
Katzung, B.G. and Payan, D.G. Non-steroidal anti-inflammatory drugs; non-opoid
analgesics; drugs used in gout, In Basic and clinical pharmacology ed. Katzung,
B.G. USA: Appleton and Lange. 1995.
Katzung, B.G. Basic and clinical pharmacology. Stanford, Connecticut: Appleton and
Lange, 1995.
Kavalier, M., Ossenkopp, K.P. and Tysdale, D.M. 1991. Evidence for the involvement
of protein kinase C in the modulation of morphine-induced ‘analgesia’ and the
inhibitory effects of exposure to 60-Hz magnetic fields in the snail, Cepaea
nemoralis, Brain Research. 554: 65-71.
Kenins, P. 1982. Response of single nerve fibers to capsaicin applied to the skin,
Neuroscience letters. 29: 83-88.
Khairatkar-Joshi, N. and Szallasi, A. 2009. TRPV1 antagonists: the challenges for
therapeutic targeting, Trends in Molecular Medicine. 15: 14–22.
Khalid, M.H., Akhtar, M.N., Mohamad, A.S., Perimal, E.K., Akira, A., Israf, D.A.,
Lajis, N. et al., 2011. Antinociceptive effect of the essential oil of Zingiber
zerumbet in mice: possible mechanisms, Jorunal of Ethnopharmacology.137:
345-351.
Khasar, S.G., Lin, Y.H., Martin, A., Dadgar, J., McMahon, T., Wang, D., et al. 1999. A
novel nociceptor signaling pathway revealed in protein kinase C epsilon mutant
mice, Neuron. 24: 253-260.
Koga, K., Honda, K., Ando, S., Harasawa, I., Kamiya, H. and Takano, Y. 2004.
Intrathecal clonidine inhibits mechanical allodynia via activation of the spinal
© COPYRIG
HT UPM
123
muscarinic M1 receptor in streptozotocin-induced diabetic mice, European
Journal of Pharmacology. 505: 75-82.
Koyama, K., Imaizumi, T., Akiba, M., Kinoshita, K., Tkahashi, K., Suzuki, A., et al.
1997. Antinociceptive components of Ganoderma lucidum, Planta Medica. 63:
224-227.
Krause, J.E., Chenard, B.L. and Cortright, D.N. 2005. Transient receptor potential ion
channels for the discovery of pain therapeutics, Current opinion in
investigational drugs. 6: 48-57.
Kreis, M.E., Jiang, W., Kirkup, A. J. and Grundy, D. 2002. Cosensitivity of vagal
mucosal afferents to histamine and 5-HT in the rat jejunum, American Journal
of Physiology - Gastrointestinal and Liver Physiology. 283: G612–G617.
Kulkarni, S.K. and Singh, V.P. 2007. Licofelone - a novel analgesic and anti-
inflammatory agent, Current topic medicinal chemistry. 7: 251-263.
Kvamme, E. 1998. Synthesis of glutamate and its regulation, Progress in brain
research. 116: 73-85.
Lavand’homme, P.M. and Eisenach, J.C. 2003. Perioperative Administration of the α2-
Adrenoceptor Agonist Clonidine at the Site of Nerve Injury Reduces the
Development of Mechanical Hypersensitivity and Modulates Local Cytokine
Expression, Pain. 105: 247–254.
Lawson, K. 1996. Potassium channel activation: a potential therapeutic approach?
Pharmacology and Therapeutics. 70: 39–63.
Le Bars, D., Gozariu, M., and Cadden, S.W. 2001. Animal Models of Nociception,
Pharmacological Reviews. 53: 597-652.
Lee, J.L., Mukhtar, H., Bickers, D.R., Kopelovich, L. and Athar, M. 2003.
Cyclooxygenase in the skin: pharmacological and toxicological implications,
Toxicology and applied pharmacology. 192: 294-306.
Lima, F.O., Souza, G.R., Verri, Jr., W.A., Parada, C.A., Ferreira, S.H., Cunha, F.Q., et
al., 2010. Direct blockade of inflammatory hypernociception by peripheral A1
adenosine receptors: involvement of the NO/cGMP/PKG/KATP signaling
pathway, Pain. 151: 506-515.
Lloyd, G.K. and Williams, M., 2000. Neuronal nicotinic acetylcholine receptors as
novel drug targets, Journal of Pharmacology and Experimental Therapeutics.
292: 461-467.
Lopez-Avila, A., Coffeen, U., Ortega-Legaspi, J.M., del Angel, R. and Pellicer, F.
2004. Dopamine and NMDA systems modulate long-term nociception in the rat
anterior cingulate cortex, Pain. 111: 136–143.
Madi, L., Bar-Yehuda, S., Barer, F., Ardon, E., Ochaion, A. and Fishman, P. 2003. A3
adenosine receptor activation in melanoma cells: association between receptor
© COPYRIG
HT UPM
124
fate and tumor growth inhibition, The Journal of Biological Chemistry. 278:
42121–42130.
Malmberg, A.B., Chen, C., Tonegawa, S. and Basbaum, A.I. 1997. Preserved acute
pain and reduced neuropathic pain in mice lacking PKC gamma. Science 278:
279–283.
Mandadi, S., Tominaga, T., Numazaki, M., Murayama, N., Saito, N., Armati, P.J., et al.
2006. Increased sensitivity of desensitized TRPV1 by PMA occurs through
PKCε-mediated phosphorylation at S800. Pain. 123: 106–116.
Mantyh, P.W. and Hunt, S.P. 2004. Setting the tone: superficial dorsal horn projection
neurons regulate pain sensitivity, Trends in Neurosciences. 27: 582-584.
Mao, J., Price, D.D., Hayes, R.L., Lu, J. and Mayer, D.J. 1992. Differential roles of
NMDA and non-NMDA receptor activation in induction and maintenance of
thermal hyperalgesia in rats with painful peripheral mononeuropathy, Brain
Research. 598: 271-278.
Marcondes Sari, M.H., Guerra Souza, A.C., Rosa, S.G., Souza, D., Rodrigues, O.E.D.
and Nogueira, C.W. 2014. Contribution of dopaminergic and adenosinergic
systems in the antinociceotive effect of p-cholor-selenosteroid, Europeand
journal of pharmacology. 725: 79-86.
Marieb, E.N. Human Anatomy and Physiology. San Francisco: Pearson Education, Inc.
2004.
Martin, E.A. Oxford Concise Medical Dictionary. Oxford: Market House Book Ltd.
2003.
Martin, W.R., Eades, C.G., Thompson, J.A., Huppler, R.E. and Gilbert, P.E. 1976. The
effects of morphine- and nalorphine-like drugs in the nondependent and
morphine-dependent chronic spinal dog, The Journal of Pharmacology and
Experimental Therapeutics. 197: 517-532.
Mayer, B., Brunner, F., and Schmidt, K. 1993. Inhibition of nitric oxide synthesis by
methylene blue, Biochemical pharmacology. 45: 367-374.
Mayer, S., Izydorczyk, I., Reeh, P.W. and Grubb, B.D. 2007. Bradykinin-induced
nociceptor sensitisation to heat depends on cox-1 and cox-2 in isolated rat skin,
Pain. 130: 14-24.
McKenna, M.C. 2007. The glutamate-glutamine cycle is not stoichiometric; fates of
glutamate in brain, Journal of Neuroscience research. 85: 3347-3358.
Meller, S.T. and Gebhart, G.F. 1993. Nitric oxide (NO) and nociceptive processing in
the spinal cord, Pain. 52: 127–136.
Merskey, N. and Bogduk, N. IASP Pain Terminology: Classification of Chronic Pain,
IASP Taskforce on Taxonomy. Seattle: IASP Press. 1994.
© COPYRIG
HT UPM
125
Millan, M.J. 1999. The induction of pain: an integrative review, Progress in
Neurobiology. 57:1-164.
Millan, M.J. 2002. Descending control of pain, Progress in Neurobiology. 66: 355-474.
Miller, K.E., Hoffman, E.M., Sutharshan, M. and Schechter, R. 2011. Glutamate
pharmacology and metabolism in peripheral primary afferents: Physiological
and pathophysiological mechanisms, Pharmacology and Therapeutics. 130:
283-309.
Miller, K.E., Richards, B.A. and Kriebel, R.M. 2002. Glutamine-, glutamine
synthetase-, glutamate dehydrogenase-, pyruvate carboxylase-
immunoreactivities in the rat dorsal root ganglion and peripheral nerve. Brain
Research. 945: 202-211.
Mohd. Sani, M.H., Zakaria, Z.A., Balan, T., Teh, L.K., Salleh, M.Z., 2012.
Antinociceptive Activity of Methanol Extract of Muntingia calabura Leaves
and the Mechanisms of Action Involved. Evidence-Based Complementary and
Alternative Medicine. 2012 10 pages. doi:10.1155/2012/890361.
Morton, J.F. 1987. Jamaica cherry in Fruits of Warm Climates ed. Morton, J.F. Miami,
FL. 1987.
Musa, A.M., Aliyu, A.B., Yaro, A.H., Magaji, M.G., Hassan, H.S. and Abdullahi, M.I.
2009. Preliminary phytochemical, analgesic and anti-inflammatory studies of
the methanol extract of Anisopus mannii in rodents, African Journal of
Pharmacy and Pharmacology. 3: 374-378.
Mycek, M.J., Harvey, R.A. and Champe P.C. Lippincott’s Illustrated Review:
Pharmacology. Philadelphia: Lippicott- Raven. 1997.
Naguib, M. and Yaksh, T.L. 1997. Characterization of muscarinic receptor subtypes
that mediate antinociception in the rat spinal cord, Anesthesiology and
Analgesia. 85: 847-853.
Nascimento, F.P., Macedo, Jr. S.J. and Santos, A.R.S. The Involvement of Purinergic
System in Pain: Adenosine Receptors and Inosine as Pharmacological Tools in
Future Treatments in Pharmacology ed. Luca Gallelli. Croatia: InTechopen.
2012. doi: 10.5772/33754.
Narita, M., Mizoguchi, H., Kampine, J.p. and Tseng, L.F. 1996. Role of protein kinase
C in desensitization of spinal delta-opioid-mediated antinociception in the
mouse, British Journal of Pharmacology. 118: 1829-1835.
Narita, M. and Tseng, L.F. 1995. Stimulation of spinal delta-opioid receptors in mice
selectively enhances the attenuation of delta-opioid receptor-mediated
antinociception by antisence oligodeoxynucleotide, European Journal of
Pharmacology. 284: 185-189.
Neugebauer, V. 2002. Metabotropic glutamate receptors-important modulators of
nociception and pain behavior, Pain. 98: 1-8.
© COPYRIG
HT UPM
126
Newman, D.J. and Cragg, G.M. 2007. Natural products as sources of new drugs over
the last 25 years, Journal of Natural Products. 70: 461-477.
Nivethetha, M., Jayasari, J. and Brindha, P. 2009. Effects of Muntingia calabura L. on
isoproterenol-induced myocardial infarction, Singapore Medical Journal.
50:300–302.
Numazaki, M., Tominaga, T., Toyooka, H. and Tominaga, M. Direct phosphorylation
of capsaicin receptor VR1 by protein kinase Cepsilon and identification of two
target serine residues, The Journal of Biological Chemistry. 277: 13375–13378.
Ocana, M., Del Pozo, E., Barrios, M., Robles, L.I. and Baeyens, J.M. 1990. An ATP-
dependent potassium channel blocker antagonizes morphine analgesia.
European Journal of Pharmacology. 186: 377–378.
Ochoa, J. and Torebjork, E. 1983. Sensations evoked by intraneural microstimulation
of single mechanoreceptor units innervating the human hand, Journal of
Physiology. 342: 633-654.
Ochoa, J. and Torebjork, E. 1989. Sensations evoked by intraneural microstimulation
of C nociceptor fibers in human skin nerves, Journal of Physiology. 415: 583-
599.
OECD principles on good laboratory practice in Handbook, Good Laboratory Practice
(GLP), Quality Practices for Regulated Non Clinical Research and Development
TDR PRD/GLP/01.2, 2001.
Ohana, G., Bar-Yehuda, S., Arich, A., Madi, L., Dreznick, Z., Rath-Wolfson, L. et al.
2003. Inhibition of primary colon carcinoma growth and liver metastasis by the
A3 adenosine receptor agonist CF101. British Journal of Cancer. 89: 1552–
1558.
Olah, Z., Karai, L. and Iadarola, M.J. 2002. Protein kinase Cα is required for vanilloid
receptor 1 activation. Evidence for multiple signaling pathways. The Journal of
Biological Chemistry. 277: 35752–35759.
Olivier, B. 2015. Serotonin: A never-ending story. European Journal of Pharmacology.
753: 2-18.
Pan, H.L., Wu, Z.Z., Zhou, H.Y., Chen, S.R., Zhang, H.M. and Li, D.P. 2008.
Modulation of pain transmission by G-protein-coupled receptors, Pharmacology
and Therapeutics. 117: 141-161.
Paris, P.M., and Stewart, R.D. Pain Management in Emergency Medicine. Connecticut:
Appleton and Lange. 1988.
Patil, C.S. Jain, N.K., Singh, A. and Kulkarni, S.K. 2003. Modulatory Effect of
Cyclooxygenase Inhibitors on Sildenafil-Induced Antinociception,
Pharmacology. 69: 183–189.
© COPYRIG
HT UPM
127
Perez-Arbelaez, E. Plantas Medicinales Y Venenosas De Colombia: Estudio Botánico,
éTnico, Farmacéutico, Veterinario Y Forense. Colombia: Hernando Salazar,
1975.
Peters, J.H., McDougall, S.J., Fawley, J.A., Smith, S.M. and Andresen, M.C. 2010.
Primary Afferent Activation of Thermosensitive TRPV1 Triggers Asynchronous
Glutamate Release at Central Neurons, Neuron. 65: 657–669.
Petrenko, A.B., Yamakura, T., Baba, H. and Shimoji, K. 2003. The role of N-methyl-
D-aspartate (NMDA) receptors in pain: a review, Anesthesia and Analgesia. 97:
1108-1116.
Pini, L.A., Vitale, G., Ottani, A. and Sandrini, M. 1997. Naloxone reversible
antinociception by paracetamol in the rat, Journal of Pharmacology and
Experimental Therapeutics. 280: 934–940.
Preethi, K., Premasudha, P. and Keerthana, K. 2012. Anti-inflammatory activity of
Muntingia calabura fruits, Pharmacognosy Journal. 4: 51–6.
Preethi, K., Vijayalakshmi, N., Shamna, R. and Sasikumar, J.M. 2010. In vitro
antioxidant activity of extracts from fruits of Muntingia calabura Linn. from
India, Pharmacognosy Journal. 2: 11–18.
Raffa, R.B. and Codd, E.E. 1994. Lack of glibenclamide or TEA affinity for opioid
receptors: further evidence for in vivo modulation of antinociception at K+
channels, Brain Research. 650: 146–148.
Raj, P. Pain Medicine: A Comprehensive Review. Texas: Mosby Inc. 2003.
Raskin, I. and Ripoll, C. 2004. Can an apple a day keep the doctor away? Current
pharmaceutical design. 10: 3419-3429.
Rasooli, I. Bioactive compounds in phytomedicine. Croatia: Intechopen, 2011.
Reddy, S.V.R. and Yaksh, T.L. 1980. Spinal noradrenergic terminal system mediates
antinociception, Brain research. 189: 391-401.
Riley, J. and Boulis, N.M. 2006. Molecular mechanisms of pain: a basis for chronic
pain and therapeutic approaches based on the cell and the gene, Clinical
Neurosurgery. 53: 77–97.
Rittner, H.L., Machelska, H. and Stein, C. Immune System, Pain and Analgesia in
Science of Pain ed. Basbaum, A.I. and Bushnell, M.C. San Diego.Elsevier Inc.,
2009.
Rodrigues, A.R. and Duarte, I.D. 2000. The peripheral antinociceptive effect induced
by morphine is associated with ATP-sensitive K+ channels, British Journal of
Pharmacology. 129: 110–114.
Rowbotham, D.J. and Macintyre, P.E. Clinical Pain Management: Acute Pain. London:
Arnold. 2003.
© COPYRIG
HT UPM
128
Rudy, B. 1988. Diversity and ubiquity of K channels, Neuroscience. 25: 729-749.
Safayhi, H. and Sailer, E.R. 1997. Anti-infammatory actions of pentacyclic triterpenes.
Planta Medica. 63: 487-493.
Sanchez de Medina, F., Romero, J.A.., Galvez, J. and Zarzuelo, A. 1996. Effect of
quercitrin on acute and chronic experimental colitis in the rat, Journal of
Pharmacology and Experimental Therapeutics. 278: 771–779.
Sanchez de Medina, F., Vera, B., Galvez, J. and Zarzuelo, A. 2002. Effect of quercitrin
on the early stages of hapten-induced colonic inflammation in the rat, Life
Sciences. 70: 3097-3108.
Sandkuhler, J. 2000. Learning and memory in pain pathways, Pain. 88: 113-118.
Sang, C.N., Gracely, R.H., Max, M.B. and Bennet, G.J. 1996. Capsaicin-evoked
mechanical allodynia and hyperalgesia cross nerve territories. Evidence for
central mechanism, Anesthesiology. 85: 491-496.
Sauer, S.K., Schafer, D., Kress, M. and Reeh, P.W. 1998. Stimulated prostaglandin E2
release from rat skin, in vitro, Life Science. 62: 2045–2055.
Sawynok, J, and Liu, X.J. 2003. The formalin test: Characteristics and usefulness of
the model. Reviews in Analgesia. 7: 145-163.
Sawynok, J. 1998. Adenosine receptor activation and nociception, European Journal of
Pharmacology. 347: 1-11.
Sawynok, J. 2011. Caffeine and pain. Pain. 152: 726–729.
Sawynok, J. and Reid, A. 1996. Interactions of descending serotonergic systems with
other neurotransmitters in the modulation of nociception, Behavioural Brain
Research. 73: 63-68.
Schaible, H.G., Schmelz, M. and Tegeder, I. 2006. Pathophysiology and treatment of
pain in joint disease, Advance drug delivery reviews. 58: 323-342.
Schidmt, B. L., Tambeli, C.H., Levine, J.D. and Gear, R.W. 2002. mu/cooperativity
and opposing opioid effects in nucleus accumbens mediated antinociception in
the rat, European Journal of Neuroscience. 15: 861-868.
Schmidt, R.F. and Willis, W.D. Encyclopedia of pain. New York: Springer-verlag
Berlin Heilberg, 2007.
Sebastião, A.M. and Ribeiro, J.A. 2009. Adenosine receptors and central nervous sys-
tem, European Journal of Pharmacology. 623: 41–46.
Serhan, C.N. and Haeggstrom, J.Z. Lipid mediators in acute inflammation and
resolution: eicosanoids, PAF, resolvins and proteins. In fundamentals of
© COPYRIG
HT UPM
129
inflammation ed. Serhan, C.N. Cambridge, U.K: Cambridge University press.
2010.
Shih, C.D. 2009. Activation of nitric oxide/cGMP/PKG signaling cascade mediates
antihypertensive effects of Muntingia calabura in anesthetized spontaneously
hypertensive rats, American Journal of Chinese Medicine. 37: 1045–1058.
Shih, C.D., Chen, J.J. and Lee, H.H. 2006. Activation of nitric oxide signaling pathway
mediates hypotensive effect of Muntingia calabura L. (Tiliaceae) leaf extract,
American Journal of Chinese Medicine. 34: 857–72.
Shu, Y.Z. 1998. Recent natural products based drug development: a pharmaceutical
industry perspective, Journal of Natural Products. 61: 1053-1071.
Sibi, G., Naveen, R. Dhananjaya, K., Ravikumar, K.R. and Mallesha, H. 2012.
Potential use of Muntingia calabura L. extracts against human and plant
pathogens, Pharmacognosy Journal. 4: 44–47.
Siddiqua, A., Premakumari, K.B. and Sultana, R. 2010. Antioxidant activity and
estimation of total phenolic content of Muntingia calabura by colorimetry,
International Journal of ChemTech Research. 2: 205–208.
Smith, J.A., Davis, C.L. and Burgess, G.M. 2000. Prostaglandin E2-induced
sensitization of bradykinin-evoked responses in rat dorsal root ganglion neurons
is mediated by cAMP-dependent protein kinase A, European Journal of
Neuroscience. 12: 3250–3258.
Smith, R.M., 2003. Before the injection--modern methods of sample preparation for
separation techniques, Journal of Chromatography A. 1000: 3–27
Soares, A.C., Leite, R., Tatsuo, M.A. and Duarte, I.D. 2000. Activation of ATP-
sensitive K+ channels: mechanism of peripheral antinociceptive action of the
nitric oxide donor, sodium nitroprusside, European Journal of Pharmacology.
400: 67–71.
Sommer, C. 2004. Serotonin in pain and analgesia, Molecular Neurobiology. 30: 117-
125.
Sommer, C. 2010. Serotonin in pain and pain control, Handbook of the behavioral
neuroscience. 21: 457-471.
Sousa, A.M. and Prado, W.A. 2001. The dual effect of a nitric oxide donor in
nociception, Brain Research. 897: 9–19.
Spano, M.S., Ellgren, M., Wang, X. and Hurd, Y.L. 2007. Prenatal Cannabis Exposure
Increases Heroin Seeking with Allostatic Changes in Limbic Enkephalin
Systems in Adulthood, Biological Psychiatry. 61: 554-563.
Sridhar, M., Thirupathi, K., Chaitanya, G., Ravi Kumar, B. and Krishna Mohan, G.
2011. Antidiabetic effect of leaves of Muntingia calabura L., in normal and
© COPYRIG
HT UPM
130
alloxan-induced diabetic rats, Journal of Pharmacology and
Pharmacotherapeutics. 2: 626–32.
Starec, M., Waitzov’a, D. and Elis, J. 1988. Evaluation of the analgesic effect of RG-
tannin using the ‘‘hot plate’’ and ‘‘tail flick’’ method in mice (in Czech), Cesk
Farm. 37: 319–321.
Steinbush, H.M.W. 1981. Distribution of serotonin-immunoreactivity in the central
nervous system of the rat-cell bodies and terminals, Neuroscience. 4: 557–618.
Steven, M.F. Chronic pain in small animal medicine. Manson Publishing (UK): The
Veterinary Press. 2010.
Strong, J., Unruh, A.M., Wright, A., Baxter, G.D. and Wall, P.D. Pain: A textbook for
therapists. London: Churchill Livingstone. 2002.
Su, N., Jung Park, E., Vigo, J.S., Graham, J.G., Cabieses, F., Fong, H.H., et al. 2003.
Activity-guided isolation of the chemical constituents of Muntingia calabura
using a quinine reductase induction assay, Phytochemistry. 63: 335–341.
Sufian, A.S., Ramasamya, K., Ahmat, N., Zakaria, Z.A. 2013. Isolation and
identification of antibacterial and cytotoxic compounds from the leaves of
Muntingia calabura L., Journal of Ethnopharmacology. 146: 198–204.
Suresh, K.K. 2012. Pharmacognostic evaluation, in vitro antioxidant and in vivo anti-
inflammatory studies of Muntingia calabura Linn., Journal of Global Trends in
Pharmaceutical sciences. 3: 805–811.
Svendsen, P., and Hau, J. Handbook of Laboratory Animal Science: Selection and
Handling of Animals in Biomedical Research, Boca Raton: CRC Press Inc.
1994.
Szolcsanyi, J. Actions of capsaicin on sensory neurons in Capsaicin in the study of pain
ed Wood, J.N. London, England: Academic press. 1993.
Taiwo, Y.O. and Levine, J.D. 1990. Direct cutaneous hyperalgesia induced by
adenosine, Neuroscience. 38: 757-762.
Talarek, S. and Fidecka, S. 2002. Role of nitric oxide in benzodiazepines-induced
antinociception in mice, Polish Journal of Pharmacology. 54: 27–34.
Tata, A.M., Vilaró, M.T. and Mengod, G. 2000. Muscarinic receptor subtypes
expression in rat and chick dorsal root ganglia, Molecular Brain Research. 82:
1-10.
Terenius, L. Families of opioid peptides and classes of opioid receptors. In Advances in
Pain Research and Therapy ed. Fields H.L, Dubner, R., and Cervero, F. New
York Raven. 1985.
Tjolsen, A., Berge, O.G., Hunskaar, S., Rosland, J.H. and Hole, K. 1992. The formalin
test: an evaluation of the method, Pain. 51: 5-17.
© COPYRIG
HT UPM
131
Treede, R.D. and Apkarian, A.V. Nociceptive processing in the cerebral cortex in
Science of Pain ed. Basbaum, A.I. and Bushnell, M.C. San Diego: Elsevier.
2009.
Treede, R.D., Meyer, R.A., Raja, S.N. and Campbell, J.N. 1992. Peripheral and central
mechanisms of cutaneous hyperalgesia, Progress in Neurobiology. 38: 397-421.
Tseng, L., Yu, J. and Pieper, G. 1992. Increase of nitric oxide production by L-arginine
potentiates i.c.v. administered beta-endorphin-induced antinociception in the
mouse, European Journal of Pharmacology. 212: 301–303.
Tsimogiannis, D., Samiotaki, M., Panayotou G. and Oreopoulou, V. 2007.
Characterization of Flavonoid Subgroups and Hydroxy Substitution by HPLC-
MS/MS, Molecules. 12: 593-606.
Turner, P.V., Brabb, T., Pekow, C., and Vasbinder, M.A. 2011. Administration of
substances to laboratory animals: routes of administration and factors to
consider, Journal of American Association Laboratory Animal Science. 50:
600–613.
Urban, L. Thompson, S.W.N. and Dray, A. 1994. Modulation of spinal excitability: co-
operation between neurokinin and excitatory amino acid neurotransmitters,
Trend in neurosciences. 17: 432-438.
Valencia, E., Feria, M., Dias, J.G., Gonzalez, A. and Bermejo, J. 1994.
Antinociceptive, antiinfammatory and antipyretic effects of lapidin, a bicyclic
sesquiterpene, Planta Medica. 60: 395-399.
Vane, J.R. 1971. Inhibition of prostaglandin synthesis as a mechanism of action for
aspirin-like drugs, Nature: New Biology. 231: 232–235.
Vane, J.R. and Botting, R.M. 2003. The mechanism of action of aspirin, Thrombosis
research. 110: 255-258.
Vanegas, H. and Schaible, H.G. 2000. Effects of antagonists to high-threshold calcium
channels upon spinal mechanism of pain, hyperalgesia and allodynia, Pain. 85:
9-18.
Vanegas, H. and Schaible, H.G. NMDA receptors in spinal nociceptive processing in
Encyclopedia of pain ed. Schmidt, R.F. and Willis, W.D. New York: Springer-
verlag Berlin Heilberg. 2007.
Vanegas, H. and Schaible, H.G. N-methyl0D-aspartate (NMDA) antagonist in
Encyclopedia of pain ed. Schmidt, R.F. and Willis, W.D. New York: Springer-
verlag Berlin Heilberg. 2007a.
Vasudevan, M. Gunnam, K.K. and Parle, M. 2006. Antinociceptive and anti-
inflammatory properties of Daucus carota seeds extract, Journal of Health
Science. 52: 598–606.
© COPYRIG
HT UPM
132
Velazquez, K.T., Mohammad, H. and Sweitzer, S.M. 2007. Protein kinase C in pain:
involvement of multiple isoforms, Pharmacological research. 55: 578-589.
Vellani, V., Mappleback, S., Moriondo, A., Davis, J.B. and McNaughton, P.A. 2001.
Protein kinase C activation gating of the vanilloid receptor VR1 by capsaicin,
protons, heat and anandamide, Journal of Physiology. 560: 391-401.
Vellani, V., Mapplebeck, S., Moriondo, A., Davis, J.B. and McNaughton, P.A. 2001.
Protein kinase C activation potentiates gating of the vanilloid receptor VR1 by
capsaicin, protons, heat and anandamide, Journal of Physiology (Lond.). 543:
813–825.
Vergana, C., latorre, R., Marrison, N.V. and Adelman, J.P. 1998. Calcium-activated
potassium channels, Current opinion in neurobiology. 8: 321-329.
Verma, P.R., Joharapurkar, A.A., Chatpalliwar, V.A. and Asnani, A.J. 2005.
Antinociceptive activity of alcoholic extract of Hemidesmus indicus R.Br. in
mice, Journal of Ethnopharmacology, 102: 298–301.
Vivancos, G.G. Parada, C.A. and Ferreira, S.H. 2003. Opposite nociceptive effects of
the arginine/NO/cGMP pathway stimulation in dermal and subcutaneous
tissues, British Journal of Pharmacology. 138: 1351–1357.
Vogel, H.G. Drug discovery and evaluation: Pharmacological Assays. J.A. Majors
Company, Lewisville, 1997.
von Frey, M. 1895. Beitrage zur sinnesphysiologie der haut, Ber. Sachs. Ges. Wiss. 47:
166–184.
Vongtau, H.O., Abbah, J., Mosugu, O., Chindo, B.A., Ngazal, I.E., Salawu, A.O., et al.
2004. Antinociceptive profile of the methanolic extract of Neorautanenia mitis
root in rats and mice, Journal of Ethnopharmacology. 92: 317-324.
Vongtau, H.O., Abbah, J., Ngazal, I.E., Kunle, O.F., Chinbo, B.A., Otsapa, P.B. and
Gamaniel, K.S. 2004. Antinociceptive and anti-inflammatory activities of the
methanolic extract of Parinari polyandra stem bark in rats and mice, Journal of
Ethnopharmacology. 90: 115-121.
Weisenberg M. Pain: Clinical and Experimental Perspective. Saint Louis: C.V. Mosby
Company. 1975.
Willis, W.D. and Coggeshall, R.E. Sensory mechanisms. The spinal cord. New York:
Plenum Press. 1991.
Willis, W.D. Spinothalamic tract neurons, role of nitric oxide. In Encyclopedia of pain
ed. Schmidt, R.F. and Willis, W.D. New York: Springer-verlag Berlin Heilberg.
2007.
Wilson, S.G., Bryant, C.D., and Lariviere, W.R. 2003. The heritability of
antinociception: Pharmacogenetic mediation of three over-the-counter
© COPYRIG
HT UPM
133
analgesics in mice, Journal of Pharmacology and Experimental Therapeutics.
305: 755-764.
Woolf, C.J. 1996. Windup and central sensitization are not equivalent, Pain. 66: 105-
108.
Woolf, C.J. and Salter, M.W. 2000. Neuronal plasticity: increasing the gain in pain,
Science. 288: 1765-1768.
Wu, W.P., Hao, J.X., Halldner, L., Lövdahl, C., DeLander, G.E., Wiesenfeld-Hallin, Z.
et al. 2005. Increased nociceptive response in mice lacking the adenosine A1
receptor, Pain. 113: 395-404.
Xie, W. Assessment of pain in animals. In Animal models of pain ed. Ma, C. and
Zhang, J.M. New York: Human Press, Springer. 2011.
Yaksh, T.L. Central pharmacology of nociceptive transmission in Textbook of pain ed.
Wall, P.D., and Melzack, R. Edinburgh: Churchill Livingston. 1999.
Yalcin, I. and Aksu, F. 2005. Involvement of potassium channels and nitric oxide in
tramadol antinociception, Pharmacology Biochemistry and Behavior. 80: 69–
75.
Yasunaka, K., Abe, F., Nagayama, A., Okabe, H. Lozada-Perez, L. Lopez-Villafranco,
E. et al. 2005. Antibacterial activity of crude extracts from Mexican medicinal
plants and purified coumarins and xanthones, Journal of Ethnopharmacology.
97: 293–299.
Zakaria, Z.A. 2007. Free radical scavenging activity of some plants available in
Malaysia, Iranian Journal of Pharmacology and Therapeutics. 6: 87–91.
Zakaria, Z.A., Fatimah, C.A., Mat, Jais A.M., Zaiton, H., Henie, E.F.P., Sulaiman,
M.R., et al. 2006b. The in vitro antibacterial activity of Muntingia calabura
extracts, International Journal of Pharmacology. 2: 290–293.
Zakaria, Z.A., Hassan, M.H., Nurul Aqmar, M.N., Mohd Zaid, S.N., Abd Ghani, M.
Sulaiman, M.R. et al. 2007d. Effects of various nonopioid receptor antagonists
on the antinociceptive activity of Muntingia calabura extracts in mice, Methods
and Findings in Experimental and Clinical Pharmacology. 29: 515–520.
Zakaria, Z.A., Mat Jais, A.M., Mastura, M., Mat Jusoh, S.H., Mohamed, A.M., Mohd.
Jamil, N.S. et al. 2007a. In vitro antistaphylococcal activity of the extracts of
several neglected plants in Malaysia, International Journal of Pharmacology. 3:
428-431.
Zakaria, Z.A., Mohamed, A.M., Mohd. Jamil, N.S., Rofiee, M.S., Hussain, M.K.,
Sulaiman, M.R., et al., 2011. In vitro antiproliferative and antioxidant activities
of the extracts of Muntingia calabura leaves, The American Journal of Chinese
Medicine. 39: 183-200.
© COPYRIG
HT UPM
134
Zakaria, Z.A., Mohd. Nor Hazalin, N.A., Mohd. Zaid, S.N.H., Abd. Ghani, M., Hassan,
M.H., Gopalan, H.K. 2007b. Antinociceptive, anti-inflammatory and antipyretic
effects of Muntingia calabura aqueous extract in animal models, Journal of
Natural Medicines. 61: 443–448.
Zakaria, Z.A., Mustapha, S., Sulaiman, M.R., Mat Jais, A.M, Somchit, M.N. and
Abdullah, F.C. 2007c. The Antinociceptive Action of Aqueous Extract from
Muntingia calabura Leaves: The Role of Opioid Receptors, Medical Principle
and Practice. 16: 130-136.
Zakaria, Z.A., Raden Mohd Nor, R.N., Hanan Kumar, G., Abdul Ghani, Z.D.,
Sulaiman M.R., Rathna Devi, G. et al. 2006. Antinociceptive, anti-inflammatory
and anti-pyretic properties of Melastoma malabathricum leaves aqueous extract
in experimental animals, Canadian Journal of Physiology and Pharmacology.
84: 1291–1299.
Zakaria, Z.A., Somchit, M.N., Sulaiman, M.R., Mat Jais A.M. and Fatimah, C.A. 2008.
Effects of various receptor antagonists, pH and enzymes on Muntingia calabura
antinociception in mice, Research Journal of Pharmacology. 2: 31–37.
Zakaria, Z.A., Sufian, A.S., Ramasamy, K., Ahmat, N., Sulaiman, M.R., Arifah, A. K.,
et al. 2010. In vitro antimicrobial activity of Muntigia calabura extracts and
fractions, African Journal of Microbiology Research. 4: 304–308.
Zakaria, Z.A., Sulaiman, M.R., Jais, A.M.M., Somchit, M.N., Javaraman, K.V. and
Abdullah, F.C. 2006a. The antinociceptive activity of Muntingia calabura
aqueous extract and the involvement of L-arginine/nitric oxide/cyclic guanosine
monophosphate pathway in its observed activity in mice, Fundamental and
Clinical Pharmacology. 20: 365–72.
Zeilhofer, H.U., Kress, M. and Swandulla, D. 1997. Fractional Ca2+
currents through
capsaicin- and proton-activated ion channels in rat dorsal root ganglia neurons,
Journal of Physiology. 503: 67-78.
Zeitz, K.P., Guy, N., Malmberg, A.B., Dirajlal, S., Martin, W.J., Sun, L., et al. 2002.
The 5-HT3 subtype of serotonin receptor contributes to nociceptive processing
via a novel subset of myelinated and unmyelinated nociceptors. Journal of
Neuroscience. 22: 1010-1019.
Zhang, L.J., Wang, X.J. and Han, J.S. 1990. Phorbol ester suppression of opioid
analgesia in rats, Life Science. 47: 1775-1782.
Zimmermann, M. 1983. Ethical guidelines for investigations of experimental pain in
conscious animals, Pain. 16: 109–110.
© COPYRIG
HT UPM
137
BIODATA OF STUDENT
Mohd.Hijaz Mohd Sani was born in Kota Kinabalu, Sabah, Malaysia on April 14th
,
1987. He received his early primary education at Sekolah Kebangsaan Malawa,
Telipok (1994-1999). He then continued his secondary education at Sekolah
Menengah St. Peter, Telipok from 200-2004 before being accepted into Labuan
Matriculation College (2005-2006). In July 2006, his application to further his study in
bachelor degree was accepted by Universiti Putra Malaysia. He completed his degree in
Biomedical Sciences in four years. During the study period, his passion and interest
towards research particularly in medicine field was developed, which encouraged him
to further his study. He pursues his study in Master of Science (Pharmacology) at the
same university. After a year, he managed to complete reasonable amount of his
research which allowed him to publish his first scientific journal. Following this, his
determination prompts him to apply for conversion from master to doctor of
philosophy which was successful.
© COPYRIG
HT UPM
138
LIST OF PUBLICATIONS
Journal:
M.H. Mohd. Sani, Z.A. Zakaria, T. Balan, L.K. Teh and M.Z. Salleh (2012).
Antinociceptive Activity of Methanol Extract of Muntingia calabura Leaves and
the Mechanisms of Action Involved. Evidence-based Complementary and
Alternative Medicine. Article ID 890361. DOI:10.1155/2012/890361
Z.A. Zakaria, M.H.M. Sani, M.N.H. Abdullah, A. Zuraini, A. Abdul Kader, L.K. Teh
and M.Z. Salleh (2014). Antinociceptive Activity of Methanolic Extract of
Muntingia calabura Leaves: Further Elucidation of the Possible Mechanisms.
BMC Complementary and Alternative Medicine. 14: 63.
Z.A. Zakaria M.H.M Sani, A.A. Kadir, L.K. Teh, M.Z. Salleh, (2016). Antinociceptive
effect of semi-purified petroleum ether partition of Muntinia calabura leaves.
Sociedade Brasileira de Farmacognosia, doi: 10.1016/j.bjp.2015.12.007
Zainul Amiruddin Zakaria, Mohammed Hijaz Sani, Mohammed Fazli Mohammar,
Nurul Shulehaf Mansor, Zurina Shaameri, Teh Ley Kek et al. 2013.
Antinociceptive activity of a synthetic oxopyrrolidine-based compound,
ASH21374, and determination of its possible mechanisms. Can. J. Pharmacol.
91: 1143-1153
Mohd. Hijaz Mohd Sani, Muhammad Taher, Deny Susanti, Teh Ley Kek, Mohd. Zaki
Salleh, Zainul Amiruddin Zakaria. 2014. Mechanisms of α-Mangostin-induced
antinociception in a rodent model. Biological Research for Nursing. 17(1): 68-
77
© COPYRIG
HT UPM