Phytochemical analysis of The Author(s) 2012 Convolvulus ...

Article

Phytochemical analysis ofConvolvulus hystrix Vahl andits biological effects in rats

AM Rezq1, Abdulrahman L Al-Malki2,Mohamed Y Refai2, Taha A Kumosani2

and Said S Moselhy2

Abstract‘‘Convolvulus hystrix Vahl’’ is a plant that has been known to Ancient Egyptians and to Arabs and its root was usedtraditionally as a purgative. Our attention was directed to the root bark as we observed that the largest part ofthe plant is deeply impeded underground. The work plan involved testing experimental animals for theinfluence of aqueous root bark extract on carbohydrate, fat and protein metabolisms as reflected on thegrowth and relevant laboratory metabolic assessment parameters. Proximate analysis showed high percen-tages of moisture (85%) and total lipids (7.2%) and surprising amounts of elements such as barium, strontium,rhodium and tellurium (1.7 mg, 3.1 mg, 8 mg and 9.1 mg/g ash, respectively). Random serum glucose valueshowed a significant decrease in the treated female group (p < 0.05). Serum total proteins of treated femalegroup were found to be increased significantly (p < 0.001), while it was found to be decreased in the relevanttreated male group (p < 0.01), together with a significant decrease in blood urea nitrogen (p < 0.05), with asignificant increase in the serum creatinine (p < 0.05). Concerning serum fat metabolic parameters, a significantdecrease in the serum triglycerides and high-density lipoproteins (p < 0.01 and p < 0.01, respectively) werefound. We concluded that the presence of huge amount of polyphenolics such as tannins is responsible for theoverall results documented as growth retarding and antinutritional factors. The results were motivating andpointed out to the possible opening of vast areas of research in the field of natural products. We recommenda series of biochemical and pharmacological studies concerning different parts of the plant as well.

KeywordsPhytochemical, Convolvulus hystrix Vahl, toxicity, metabolism, rats

Introduction

The plant has been documented scientifically as

‘‘Convolvulus hystrix Vahl’’ and has been known to

Ancient Egyptians and to Arabs. The plant’s basic

morphology and uses were mentioned in traditional

Arabs medicine. Yet, it did not attract the attention

of scientific committee. Revising published research

together with extensive search on the ‘‘Internet’’ did

not reveal any such interest. All published data were

concerned with its historical documentation, botanical

taxonomy, morphology, regional and geographical

distribution, ‘‘this helped a lot,’’ but did not give a sin-

gle direct clue to follow, concerning the plant’s meta-

bolic effects. According to Michael (1995), there are

only few basic structural differences between the

continuously extended root and stem bark. Studies

on the morphological and anatomical structures of the

aerial parts of C. hystrix under investigation revealed

that neither biochemical nor metabolic research has

been cited and found that the separation and

1 Unit of Biochemistry and Molecular Biology, the MedicalBiochemistry Department, Faculty of Medicine, Cairo University,Cairo, Egypt2 Biochemistry Department, Faculty of Science, King AbdulazizUniversity, Jeddah, Kingdom of Saudi Arabia

Corresponding author:Said S Moselhy, Biochemistry Department, Faculty of Science,King Abdulaziz University, Jeddah, Saudi Arabia.Email: [email protected]

Toxicology and Industrial Health2015, Vol. 31(4) 304–318© The Author(s) 2012Reprints and permissions:sagepub.co.uk/journalsPermissions.navDOI: 10.1177/0748233712469653tih.sagepub.com

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identification of some active chemical constituents of

the aerial parts of this plant were carried out by Dawi-

dar et al. (2000). The work plan involves testing

experimental animals for the influence of this extract

to give to other general metabolic, hematological and

histopathological assessments as confirmatory mea-

sures to the expected metabolic influences. Dawidar

et al. (2000) identified 10 natural products, of which

a stilbene carboxylic acid was isolated for the first

time as a new natural product. The isolated com-

pounds include seven shikimates: vanillin, vanillic

acid, syringic acid, ferulic acid, isoferulic acid,

isoscopoletin and a new stilbene carboxylic acid deri-

vative, in addition to two sterols: beta-sitosterol and

stigmasterol as well as one triterpenoid (oleanolic

acid).

The phenolic fractions of the aerial part of Scro-

phularia frutescens and Scrophularia sambucifolia

(Scrophulariaceae) showed a potent antibacterial

activity. Ferulic, isovanillic, p-hydroxycinnamic,

p-hydroxybenzoic, syringic, caffeic, gentisic and pro-

tocatechuic acids were isolated from S. frutescens,

and ferulic, p-coumaric, vanillic, p-hydroxibenzoic

and syringic acids were isolated from S. sambucifolia.

Since phenolic acids have been shown in the literature

to exert an antibacterial effect, the presence of these

compounds in the two plants explains their antibacter-

ial activity (Fernandez et al., 1996). Four of the 12

phenolic compounds detected in the nonvolatile frac-

tion (caffeic acid, vanillin, syringaldehyde and ellagic

acid), the amounts of which increase during the

maturation of whisky, were found to strongly inhibit

the mouse alcohol dehydrogenases class I (Haseba

et al., 2008). Abbas et al. (2007) investigated the

aerial part of Commiphora opobalsamum L. (Burser-

aceae) growing in Saudi Arabia, in order to identify

its major chemical constituents and to evaluate its

extracts and isolated compounds for antimicrobial,

antimalarial, antitumor, anti-inflammatory cyclooxy

genase-2 (inhibition), antioxidant and estrogenic

activities. Of the isolated compounds, syringic acid

showed moderate antimalarial, anticandidal and anti-

mycobacterial activities. As an antioxidant, ferulic

acid may neutralize free radicals such as reactive oxy-

gen species that may be involved in DNA damage and

accelerated cell aging. Animal studies and in vitro

studies suggest that ferulic acid may have direct anti-

tumor activity against breast cancer and liver cancer.

Ferulic acid may have proapoptotic effects in cancer

cells, thereby leading to their destruction. Ferulic acid

has been shown to be effective in treating hot flushes

in postmenopausal women. Ohara et al. (2008) inves-

tigated the effect of hydroxycinnamic acid derivatives

(HADs) on serum adiponectin concentrations and on

adiponectin secretion of 3T3-L1 adipocytes in mice.

It was found that trans-ferulic acid markedly

enhanced the adiponectin secretion of 3T3-L1 adipo-

cytes. The results indicate that these HADs might reg-

ulate adiponectin secretion by the inhibition of

nuclear factor-kappaB activation. HADs might be

effective for ameliorating type 2 diabetes.

Materials and methods

Plant samples have been collected as one batch. The

leaves, inflorescence and flowers were removed and

kept separately at �20�C for future study. The roots

were washed lightly to remove sand then dried with

clean tissue papers, after which their bark was peeled

out and stored in batches of 500 g each at �20�C for

further processing. Frozen root bark batches were

allowed to thaw and immediately cut into small pieces

about 3 � 3 cm2 which was then blend in high-speed

laboratory blender with cold bidistilled deionized:

water (1:1, w/v). The blend was further homogenized

in a high-speed homogenizer (6000 r/min) then

squeezed out in four-layered ordinary surgical gauze

and the filtrate was collected and stored in aliquots of

80 ml in a 100-ml capacity brown glass bottles at

�20�C for future use in other parts of the study. Root

bark chemical analysis was followed adopting the

methods of the Association of American Agricultural

Chemists (AOAC, 1984) and included moisture, total

crude extract protein, total fat, total crude carbohy-

drates, crude fibers, ash and its elemental composition.

Total moisture percentage

Total moisture has been determined using a drying oven

at 105�C repeatedly on a sample of about 10 g of fresh

crude sample until a constant weight was recorded for

three times successively. Total moisture was calculated

as a percentage of fresh sample weight.

Total protein percentage

Total protein was determined according to the method

of Kjeldahl published by Burns (1984). Total protein

was calculated as a percentage of fresh sample weight.

Total fat percentage

Total fat was determined through extraction of dried

sample with petroleum ether (40–60) by the Soxhlet

Rezq et al. 305



Apparatus to constant weight. Total fat was calculated

as a percentage of fresh sample weight.

Total crude fiber percentage

The AOAC method for the determination of crude

fibers content of plant samples depends on digestion

with 0.3 N sulfuric acid followed by another digestion

with 0.3 N sodium hydroxide. The washed and dried

unhydrolyzable fraction represents the total crude

fiber of the plant material, which was then calculated

as percentage of the fresh sample.

Total ash percentage

Total ash percentage was determined by ashing a

dried sample in a muffle furnace at 550�C for 12 h.

Total ash was calculated as a percentage of fresh

sample weight.

Total carbohydrate percentage

According to the AOAC (1984), total soluble ‘‘diges-

tible’’ carbohydrates were determined by difference

as a percentage of fresh sample.

Ash elemental content percentage

Ash elemental analysis was performed using a Plasma

(inductively coupled plasma) Atomic Absorption

Spectrophotometer technique (Optima 4300DV;

Perkin-Elmer, MA, USA) and a Scanning Electron

Microscope JSM-6490LV (Joel, Tachikaua-Tokyo,

Japan), according to the method of Goldstein et al.

(1992). This included aluminum, barium, boron, cal-

cium, carbon, chlorine, chromium, cobalt, cupper, iron,

lithium, magnesium, manganese, nickel, oxygen, phos-

phorus, potassium, rhodium, sodium, strontium, tellur-

ium, titanium, vanadium, zink and zirconium.

Toxicity studies

Acute toxicity of the extracts was determined in mice

according to the method of Irwin (1961) to calculate

the dose of the extract, which kills 50% of the animals

(lethal dosage (LD50)), during the 24-h period. The

extract was administrated to the animals in various

doses and watched daily for apparent changes, such

as signs of toxicity, general condition of eyes, mouth,

posture, gait and behavior.

Chronic toxicity

The chronic toxicity was carried out on rats of both

sexes according to the method of Irwin (1961), for

1 month. The extract was administrated for 2 weeks,

and the animals were observed daily for any apparent

changes, such as signs of toxicity, general condition

of eyes, mouth, posture, gait and behavior.

Animals

A total of 48 Albino Wistar rats (140–150 g) were

obtained from the Animal House facility of King

Fahad Medical Research Centre, King Abdulaziz Uni-

versity, Jeddah, Saudi Arabia. Animals were kept in

cages in groups; each group of six under standard

laboratory conditions (temperature 25 + 2�C; photo-

period of 12 h) under standard ration (Baker, 1986).

The ration was obtained from the Grain Silos & Flour

Mills Organization (Jeddah, Saudi Arabia). Animals

were kept together during the 1-week adaptation

period before starting the experiment. The rats were

divided into four groups: group I: normal control male

group (n¼ 12), group II: normal control female group

(n¼ 12), these two groups were fed ad libtium and not

treated with root bark extract, group III: male group

treated with root bark extract (n ¼ 12) and group

IV: female group treated with root bark extract

(n ¼ 12).

Administration of root bark extract

On the basis of the results of acute and chronic toxi-

city studies mentioned above, a daily dose of 2 ml

of the root bark extract was given orally at

10:00 am by stomach gavage tube size 16 mm to all

members of the treated groups. An equal volume of

distilled water was given by the same route to the

normal control groups. All the groups were followed

up on a daily basis for food and water consumption

and body weight change, for a period of 30 days. Ani-

mals were observed daily for any apparent changes,

such as signs of toxicity, general condition of eyes,

mouth, etc. posture, gait and behavior, according to

the method of Irwin (1961).

Blood sample collection

At the end of the 30-day experimental period, from all

the animals blood samples were collected from the

retro-orbital plexus under anesthesia according to van

Herck and his group (1992). The blood samples were

divided into two portions: one in a plain tube for

306 Toxicology and Industrial Health 31(4)



serum separation and second in ethylenediaminete-

traacetic acid for hematological studies. After blood

sampling, all animals were killed by cervical disloca-

tion and liver, lung, kidney, spleen, heart and testicles

(male groups) are obtained through laparotomy.

Serum was separated by centrifugation, aspirated and

kept at�20�C until analyzed, while the internal organs

were weighted individually and suitable parts of each

organ were preserved in 10% formalin for histopatholo-

gical examination. The main part of each organ was kept

at�20�C for future biochemical analysis.

Histopathological examination

Samples of the formalin-fixed livers from all four

experimental groups were sequentially dehydrated

with increasing concentrations of ethanol after which

the tissue samples were embedded in paraffin. Micro-

tome sectioned and stained with hematoxylin and

eosin according to Lynch et al. (1969) for histopatho-

logical examination under light microscope.

Statistical analysis

Statistical analysis was performed on a PC using

Statistica, V.8, Statsoft, Inc (Victoria, Australia). Data

are presented as arithmetic mean + standard devia-

tion. Student’s t test was used for the determination

of the significance of difference between sample

means, while regression analysis was used for the

determination of the presence of statistically signifi-

cant correlations between variables.

Results

It is clear from Table 1 that the total lipid components

constitute the highest percentage of the root bark on

dry weight basis.

Surprisingly, very high levels of tellurium

(9066 mg/g ash), rhodium (8000 mg/g ash), strontium

(3060.4 mg/g ash) and barium (1736.6 mg/g ash) were

found in the ash (Table 2).

Table 3 shows statistically significant differences

in mean body weight of all four groups (p < 0.05–

0.001). It is clear that the 30-day experimental period

has affected the growth in the normal control and the

treated groups positively. Males either the normal

control or the treated one gained more weight than

females either normal control or treated. The effect

of treatment on the change in mean body weight of

males and females either normal or treated is pre-

sented in Table 4.

It is clear from Table 4 that statistically highly

significant differences (p < 0.001) have been found

after the 30-day experimental period treatment with

C. hystrix Vahl root bark extract and have affected the

growth negatively in treated groups. The males either

normal control or treated gained more weight than

females either for normal control or treated.

Time growth trend

The time trend of growth for all four experimental

groups as presented by mean body weights is shown

in Figure 1.

Table 1. Results of chemical analysis of the root bark, ingram percentage.

ParameterPercent (fresh weight)

mg/g dry weight

Moisture 85Crude proteins 0.73Total lipids 7.2Ash 0.6Total carbohydrates 0.4Crude fibers 6.1

Table 2. Elemental analysis of the root bark ash presentedas microgram per gram ash.

Elements Microgram per gram ash

Aluminum 0.97Calcium 23.24Iron 0.52Potassium 14.23Magnesium 1.47Manganese 0.046Sodium 5.06Phosphorus 0.88Titanium 0.06Boron 142.5Barium 1736.6Cobalt 2.9Chromium 1.7Cupper 70.7Lithium 11.54Nickel 21.1Strontium 3060.4Vanadium 11.5Zink 108.40Zirconium 8.85Rhodium 8000Tellurium 9066

Rezq et al. 307



Table 3. Statistical analysis of significant differences in mean body weight of animals in the four experimental groups atday 30 of the experiment.

ParameterNormal Treated

Female (n ¼ 12) Male (n ¼ 12) Female (n ¼ 12) Male (n ¼ 12)

Body weight (g), mean + SD 190.41 + 10.2 285.75 + 25.3 180.52 + 9.8 255.3 + 16.7

Normal female vs. treated female

t Test 2.3740

p Value <0.05

Normal female vs. normal male

t Test 12.1228

p Value <0.001

Treatment female vs. treated male

t Test 12.6803

p Value <0.001

Normal male vs. treated male

t Test 3.2582

p Value <0.005

Table 4. Statistical analysis of significant differences in mean body weight (g) of animals in the four experimental groups,at day 1 when compared with day 30 of the experiment.

ParameterNormal Treated

Female (n ¼ 12) Male (n ¼ 12) Female (n ¼ 12) Male (n ¼ 12)

Body weight (g), mean + SD 148.67 + 7.9 152.58 + 10.3 148.17 + 9.5 154.16 + 15.1

Normal female vs. treated female

t Test 11.2573

p Value <0.001

Normal female vs. normal male

t Test 8.0518

p Value <0.001

Treatment female vs. treated male

t Test 16.9089

p Value <0.001

Normal male vs. treated male

t Test 14.9387

p Value <0.001




As Figure 1 shows, together with special references

to Tables 2 to 4, the effect of the treatment with oral

C. hystrix Vahl root bark extract has proved of statis-

tically highly significant growth retarding effect on

mean daily body weight increase in both males and

females groups when compared with relevant control

groups. Regression analysis of the rate of change in

body weight ‘‘time trend’’ as presented by mean body

weights of all four experimental animal groups is pre-

sented in Table 4 and Figures 2 to 5. Statistically

highly significant correlations (r ¼ 0.96–0.99) have

been found between time and mean body weigh daily

change (p < 0.001) in all the groups. The regression

lines show a linear relationship obeying the first-

order equation (Y ¼ a þ bX). However, the rate of

daily change in body weights (g) as presented by the

slope of the regression lines (b) is evidently less in the

treated groups when compared with the relevant

normal control groups.

Food and water consumptions

Table 5 shows food and water consumption in all the

groups on days 1 and 30 of the experiment.

As for day 1, all statistical comparisons gave statis-

tically nonsignificant differences except for three

comparisons viz., normal control male consumed

more water than normal control females (p < 0.001),

1 3 5 7 9 11 13 15 17 19 21 23 25 27 29 31Time (day)

140

160

180

200

220

240

260

280

300M

ean

body

wei

ghts

(g)

Normal female Tretaed female Normal male Treated male

Figure 1. Time trend of the change in weight per day aspresented by mean body weights (g) for normal and treatedmale and female groups.

Normal Treated male male

Figure 2. Histopathological slide of normal female liver(�10).

Figure 3. Histopathological slide of treated female liver(�10).

Figure 4. Histopathological slide of normal male liver(�10).

Rezq et al. 309



the same applies to treated males when compared

with treated females (p < 0.001).

However, on day 30 (end of the experiment), nor-

mal control male mean food consumption was statis-

tically significantly higher than normal control

females (p < 0.01). On the other hand, treatment with

C. hystrix Vahl root bark extract has led to a statisti-

cally significant decrease in food consumption of the

treated female groups when compared with their nor-

mal counterparts (p < 0.05). The treatment did not

affect the amount of food consumed by treated males

when compared with their normal control counter-

parts, but its effect was highly reflected more on the

food consumption of the treated female groups being

statistically significantly lower than the treated male

group (p < 0.001). On comparison of water consump-

tion of normal control females and males, a statisti-

cally significantly higher water consumption was

found in the male group (p < 0.001). However, treat-

ment with oral C. hystrix Vahl root bark extract has

statistically significantly decreased water consump-

tion of the treated female when compared with their

normal control counterparts (p < 0.01). The same

applies to treated male group when compared with

their normal control counterparts (p < 0.001). On

the other hand, treated males consumed statistically

higher amount of water than treated female

(p < 0.01). The cumulative reflection of food and

water consumption on growth and body weight is

presented in Table 5, which shows a statistically

nonsignificant difference in food consumption in

either the normal or the treated female groups.

However, in the normal male group, a statistically

significant increase is found in food consumption

by the normal male in day 1 when compared with

day 30 (p < 0.05 and p < 0.001, respectively).

As concerns water consumption in all the studied

groups on day 1 when compared with day 30, a statis-

tically nonsignificant difference was noticed on all

comparisons except a statistically highly significant

decrease on day 30 when compared with day 1 in the

treated male group (p < 0.001).

Statistical regression analysis has shown some

significant linear correlations of mean daily food

and water consumption and their percentage to

mean daily body weights as shown in Table 5. The

change in daily mean body weight in the normal

control male group per gram of food consumed is

more than double that in the treated male group

(b ¼ 8.01 and 3.63, respectively). On the other

hand, the linear relationship of contribution of

mean daily water consumption to mean daily body

weight increase has been lost in the treated male

group. No other statistically significant linear

correlations comparable to the above mentioned

correlations has been found in either normal con-

trol or treated female groups.

Participations of major internal body organsweights to total body weight percentage

Tables 6 to 9 show participations of the studied

individual major internal body organs to total body

weights as a percentage together with their statistical

analysis of the four studied experimental groups, to

clarify whether there are statistically significant

differences due to either the sex of the experimental

animal or the effect of treatment with oral C. hystrix

Vahl root bark extract on the mean of some major

internal body organs.

On the other hand, the participation of some

major internal body organs’ means weight percent-

age in normal control males to females as pre-

sented in Table 6 shows statistically significant

lower participation of heart (p < 0.05), spleen

(p < 0.05) and lungs in male than that in the female

control groups (p < 0.001). The same applies on

liver and kidneys but the differences are statically

nonsignificant.

The effect of treatment with oral C. hystrix Vahl

root bark extract as reflected on mean participation

percentage of some major internal body organ

weights is presented in Table 7. Treated male showed

statistically significantly less percentage participation

Figure 5. Histopathology slide of treated male liver (�10).




Table 6. Statistical analysis of significant differences in mean of some major internal body organ to body weights percent-age of the normal control female and males at the end of the experiment.

Normal, mean + SD

Female Male

Liver % (n ¼ 11) 3.98 + 0.4 Liver %(n ¼ 12) 3.8 + 0.4

Heart % (n ¼ 11) 0.47 + 0.07 Heart % (n ¼ 12) 0.41 + 0.1

Spleen % (n ¼ 12) 0.23 + 0.03 Spleen % (n ¼ 12) 0.2 + 0.03

Lung % (n ¼ 12) 0.82 + 0.1 Lung % (n ¼ 12) 0.59 + 0.1

Kidneys % (n ¼ 12) 0.79 + 0.06 Kidneys % (n ¼ 12) 0.76 + 0.1

Liver

t Test 1.5207

p Value NS

Heart

t Test 2.3461

p Value <0.05

Spleen

t Test 2.4493

p Value <0.05

Lung

t Test 5.2792

p Value <0.001

Kidneys

t Test 1.3851

p Value NS

Table 5. Descriptive statistical analysis of mean daily food and water consumptions at the start, day 1 and at the end,day 30 of the study.

Normal Mean + SD Treated Mean + SD

Day 1 (start of the experiment)Female Female

Food consumption (g) 13.91 + 1.37 Food consumption (g) 13.66 + 1.72Water consumption (ml) 17.53 + 1.7 Water consumed (ml) 15.08 + 2.02

Male MaleFood consumption (g) 13.41 + 1.97 Food consumption (g) 14.9 + 1.9Water consumption (ml) 22.88 + 1.75 Water consumption (ml) 23.28 + 3.15

Day 30 (end of the experiment)Female Female

Food consumption (g) 15 + 1.53 Food consumption (g) 13.08 + 1.88Water consumption (ml) 19.15 + 3.08 Water consumption (ml) 15.75 + 1.09

Male MaleFood consumption (g) 17.91 + 2.84 Food consumption (g) 16.84 + 2.34Water consumption (ml) 25.1 + 3.55 Water consumption (ml) 18.83 + 2.28

Rezq et al. 311



for all studied major internal body organs when com-

pared with treated females.

Statistically nonsignificant difference in mean par-

ticipation of major internal body organ percentage in

the treated female groups when compared with the

normal female counterparts for all studied organs

except a significant decrease in the mean participation

percentage of spleen to total body weight (p < 0.01)

(Table 8).

Statistically nonsignificant difference decreases in

the mean participation of some major internal body

organs to body weight percentages in the treated male

groups was found for all studied organs except a sig-

nificant decrease in the mean participation percentage

of liver to total body weight (p < 0.05), with an excep-

tional statistically significant increase in lung weight

participation percentage (p < 0.05) (Table 9).

Histopathological examination results

Histopathological examination of the livers of the

female- and male-treated animals did not show any

significant changes when compared with the relevant

normal control groups.

Discussion

Migahed (1978) described the plant, whereas Bailey

and Danin (1981) stated that Bedwens in ‘‘Sainai’’

of Egypt use the juice of the plant root mixed with

goat’s milk as a purgative. Boulus (1983) mentioned

that C. hystrix VahI is a common wild plant growing

in the Kingdom of Saudi Arabia and used in tradi-

tional medicine as a purgative. Al-Yahya et al.

(1990) studied the anatomical characteristics of the

aerial parts of plant.

Table 7. Statistical analysis of the significant differences in some major mean internal body organ percentage of thefemales and males in treated groups at the end of the experiment.

Treated, mean + SD

Female Male

Liver % (n ¼ 12) 3.82 + 0.3 Liver % (n ¼ 11) 3.46 + 0.3

Heart % (n ¼ 12) 0.43 + 0.1 Heart % (n ¼ 11) 0.4 + 0.1

Spleen % (n ¼ 12) 0.2 + 0.004 Spleen % (n ¼ 11) 0.19 + 0.01

Lung % (n ¼ 12) 0.8 + 0.2 Lung % (n ¼ 11) 0.67 + 0.4


Liver

t Test 2.7536

p Value <0.05

Heart

t Test 1.1097

p Value <0.05

Spleen

t Test 2.3803

p Value <0.05

Lung

t Test 2.2148

p Value <0.005

Kidneys

t Test 2.8819

p Value <0.01




The search for the plant according to Migahed

(1978) and as confirmed by Boulus (1983) and later

by Colentte (1999) has led to the localization of plenty

of shrub populations around the Old Airport Area and

Madinah Road, Jeddah, Saudi Arabia. Field trips

looking for the plant were successful in its localiza-

tion additionally in King Abdulaziz University Cam-

pus, Jeddah, Saudi Arabia. Figure 9 shows the form of

several densely flourished huge groupings.

Positive identification and taxonomy of the col-

lected samples confirmed their true nature as described

and classified by Migahed (1978), Al-Yahya et al.

(1990) and Colentte (1999), curtsy of Dr Faraj H.

AL-Jamdy.

Concerning the composition of the plant under inves-

tigation, Khalil et al. (1981) and EI-Tawil (1983) have

found coumarins, alkaloids, flavonoids, sterols and tri-

terpenes in the aerial parts of the plant. Dawidar et al.

(2000) identified 10 natural products, of which a new

stilbene carboxylic acid was isolated for the first time

as a new natural product. The isolated compounds

include seven shikimates: vanillin, vanillic acid, syrin-

gic acid, ferulic acid, isoferulic acid, isoscopoletin and

a new stilbene carboxylic acid derivative, in addition

Table 8. Statistical analysis of the significant differences in some major mean internal body organ percentage of thefemales in normal control and treated groups at the end of the experiment.


Female Female

Liver %(n ¼ 11) 3.98 + 0.4 Liver % (n ¼ 12) 3.82 + 0.3

Heart % (n ¼ 11) 0.47 + 0.07 Heart % (n ¼ 12) 0.43 + 0.1

Spleen (n ¼ 12) 0.23 + 0.03 Spleen % (n ¼ 12) 0.2 + 0.004

Lung % (n ¼ 12) 0.82 + 0.1 Lung % (n ¼ 12) 0.8 + 0.2


Liver

t Test 1.5346

p Value NS

Heart

t Test 1.2906

p Value NS

Spleen

t Test 2.8599

p Value <0.01

Lung

t Test 1.2379

p Value NS

Kidneys

t Test 0.2840

p Value NS

Rezq et al. 313



to two sterols: beta-sitosterol and stigmasterol as well as

one triterpenoid (oleanolic acid).

However, in view of being experimenting with a

whole water extract that contains plenty of active nat-

ural compounds, it is not expected (logically) to attri-

bute the various effects obtained as a result of

treatment to a specific compound or a group of com-

pounds, as is the case with all crude natural extracts.

The analysis of the research results gave huge volume

of documented data that need space other than this

thesis for their discussion and publication. However,

all results have been statistically analyzed and com-

mented on to the finest detail for future reference in

separate publications.

The chemical analysis of the root bark of the plant has

shown the highest content to be 85% water, together

with 8.5% total lipids, which led to the milky juicy fluid

drainage on cutting or breaking the root. This was con-

firmed by the fact that on dry weight basis, the highest

content was for total lipids (56.7%) (Table 1).

Surprisingly, very high levels of tellurium

(9066 mg/g ash), rhodium (8000 mg/g ash), strontium

Table 9. Statistical analysis of the significant differences in some major mean internal body organ percentage of the malesin normal control and treated groups at the end of the experiment.


Male Male

Liver %(n ¼ 12) 3.8 + 0.4 Liver % (n ¼ 11) 3.46 + 0.3

Heart % (n ¼ 12) 0.41 + 0.1 Heart % (n ¼ 11) 0.4 + 0.1

Spleen (n ¼ 12) 0.2 + 0.03 Spleen % (n ¼ 11) 0.19 + 0.01

Lung % (n ¼ 12) 0.59 + 0.1 Lung (n ¼ 11) 0.67 + 0.4


Testicle (n ¼ 10) 1.29 + 0.24 Testicle (n ¼ 10) 1.35 + 0.2

Liver

t Test 2.2840

p Value <0.05

Heart

t Test 0.4069

p Value NS

Spleen

t Test 1.2988

p Value NS

Lung

t Test 2.1895

p Value <0.05

Kidneys

t Test 1.7613

p Value NS

Testacies

t Test 0.6123

p Value NS




(3060.4 mg/g ash) and barium (1736.6 mg/g ash) were

found in the ash, Table 2. This shows a clear indica-

tion of the presence of such elements in the sandy

ground from which plant sample was collected.

Whether this is due to a selective absorption of such

elements specifically by this plant or being arbitrary

for all the plants in the mentioned area should be

further investigated.

Treatment with oral C. hystrix Vahl root bark

extract has proved of statistically highly significant

growth retarding effect on mean daily body weight

increase in both male and female rats when compared

with relevant normal control groups (Tables 2 to 4).

Regression analysis of the growth ‘‘time trend’’ of all

four experimental animal groups, Table 4 and Figures

2 to 5, showed statistically highly significant correla-

tions (r ¼ 0.96–0.99) between time and daily mean

body weigh change (p < 0.001) in all the groups. The

regression lines showed a first-order linear relation-

ship with the slopes of the regression lines (b) is evi-

dently less in the treated groups when compared with

the relevant normal control groups.

The cumulative reflection of food and water con-

sumption on growth and body weight, Table 3, showed

statistically different sex-related results in either the

normal or treated groups which could only be

explained on hormonal basis of the sex differences.

Statistical regression analysis, Table 4, of the change

in daily mean body weight in the normal control male

group per gram of food consumed is more than double

that in the treated male group (b ¼ 8.01 and 3.63,

respectively). The linear relationship of contribution

of mean daily water consumption to mean daily body

weight increase has been lost in the treated male group.

The participation of some major internal body

organs’ mean weights to total body weight percentage

in normal control and treated groups. Table 5 shows

some statistically significant lower participation of

studied organs due to the effect of treatment, with

an exceptional statistically significant increase in lung

weight participation percentage, in treated males

(p < 0.05). Histopathological examinations are now

being considered to try to explain such results.

All of the above mentioned changes could be

attributed to the antinutritional effects of polypheno-

lics which have been demonstrated using laboratory

animals (Myer and Gorbet, 1985; Rostagno et al.,

1973) and humans (Hussein and Abbas, 1985; Stavric

and Matula, 1992). These effects could be manifested

as a decrease in growth rate or food metabolism

(Mehansho et al., 1985 and Myer and Gorbet, 1985;

Rostagno et al., 1973), inhibition of enzymes (Ahmed

et al., 1991; Guyot et al., 1996; Oh and Hoff, 1986)

and lower egg production (Sell et al., 1983).

The shikimates that has been identified in the plant

under investigation are the precursors of countless

phytochemicals shikimate-derived phenolics such as

tannins are widely distributed in plant kingdom

(Bruyne et al., 1999). Phenolic acids are the precur-

sors for condensed tannins in the shikimate pathway,

the levels of these compounds are positively corre-

lated. Total phenolic acids were highly correlated

with condensed tannin concentrations. According to

Dewick (1995), one way of biosynthesis of polyphe-

nols is through the transformation of shikimate phe-

nolic compounds.

C. hystrix Vahl root bark extract contains the same

shikimates identified in the aerial parts of the plant.

According to Michael (1995), there are only few

basic structural differences between the continu-

ously extended root and stem bark. The thickened

roots are storage organs that differ from true tubers.

The relationship of the aerial bark to the root bark are

well known and documented especially in the bota-

nical family, ‘‘Convolvulaceae’’ with the specially

well-known model of sweet potato ‘‘Impomoea bata-

tas’’. The total content of phenolics in seven sweet

potato tubers ranges from 117 to 467 mg/kg of fresh

weight (Walter and Purcell, 1979). Many plants of the

family ‘‘Convolvulaceae’’ are characterized by having

taproots that accumulate biologically active chemicals.

A total of 289 possible secondary metabolites were

quantified.

Condensed tannins complexes with proteins could

explain the antinutritional effects of tannin-

containing ingredients in nonruminant (Martin-

Tanguy et al., 1977) and ruminant (Kumar and Singh,

1984) feeds. Interaction of phenolics with proteins is a

well-known phenomenon (Carter et al., 1972). Diges-

tive enzymes such as trypsin, lipase and amylase are

affected (Salunkhe et al., 1992). Tannins have been

reported to interfere with pancreatic digestion (Dried-

ger and Hatfield, 1972). Polyphenols found in beans

and peas inhibit the activity of amylases in vitro and

may also be responsible for diminishing the overall

digestibility of carbohydrates in the intestinal tract

of rats (Griffith, 1979, 1981). This may be due to

phenol–enzyme interaction as well as starch–phenol

association.

As concerns carbohydrate, the results of the present

study indicate a statistically significantly lower ran-

dom mean serum glucose in the treated female group

Rezq et al. 315



than that of their normal control counterparts

(p < 0.05). In the male group, the effect of treatment

on the serum amylase activity showed a statistically

significant decrease in enzyme activity (p < 0.001),

Table 9. This could be due to pancreatic affection and

to the fact that tannins also form complexes with

certain types of carbohydrates (Cai et al., 1989). On

the other hand, no effects of the treatment were of sig-

nificant effect on initial glycolytic steps of glucose as

reflected on serum LDH activity.

Such results are in accordance with previous

reports which showed that tea polyphenols exhibit a

broad spectrum of biological activities. Green tea

polyphenols inhibit intestinal uptake of glucose

through the sodium-dependent glucose transporter of

the rabbit intestinal epithelial cells. It has been sug-

gested that tea polyphenols act as antagonist-like

molecules (Kobayashi et al., 2000) and thus may con-

tribute to the reduction of blood glucose level.

It has been shown that water extracts of bean testa

inhibit the in vitro activity of a-amylase, lipase and

trypsin (Griffith, 1979). Sorghum tannins inhibit the

activity of a-amylase. Griffith and Moseley found that

trypsin and a-amylase activities in the intestine of rats

fed on a diet containing high levels of tannins of bean

testa are significantly reduced. On the other hand,

betel tannins cause antinutritional effects such as

diminishing palatibility or inhibiting the postdigestive

metabolism.

The effect of treatment with C. hystrix Vahl root

bark extract on serum lipid profile did not show any

statistically significant differences in the female

groups. However, the profile has shown a different

pattern in the treated male when compared with its

normal control male counterparts group, as a statisti-

cally significant decrease in serum triglycerides and

high-density lipoprotein together with an increase in

low-density lipoprotein has been found.

Histopathological examination of the livers of the

female- and male-treated animals did not show any

significant changes when compared with the relevant

normal control groups. This might suggest the rela-

tively short duration of the experimental period was

not enough to produce histopathologically evident

effects. Investigation of other more sensitive organs

might reflect some effect of the treatment with the

experimental extract.

However, in all the above discussions of the huge

results outcome, it is not expected to explain every

little bit of the results. As mentioned above, the pri-

mary goal of this study was to document the effect

of treatment with oral C. hystrix Vahl root bark extract

on carbohydrates, fats and proteins metabolism, as

indicators of the general growth status which in view

of being experimenting with a whole water extract

that contains plenty of active natural compounds, it

is not expected (logically) to attribute the various

effects obtained as a result of treatment with a specific

compound or a group of compounds, as is the case

with all crude natural extracts.

Extensive readings and more experimental work

with fractionated water and organic solvent extracts

are required to complete the picture of such a novel

study.

Funding

This research received no specific grant from any funding

agency in the public, commercial, or not-for-profit sectors.

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