SOME PHARMACOLOGICAL STUDIES ON ANTIBACTERIAL DRUG ...

www.wjpps.com Vol 9, Issue 3, 2020.

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Elmehalawy et al. World Journal of Pharmacy and Pharmaceutical Sciences

SOME PHARMACOLOGICAL STUDIES ON ANTIBACTERIAL DRUG

PROTECTIVE EFFECTS OF GINSENG ON CEFTRIAXONE-

INDUCED HEPATIC INJURY IN RATS

1Ashraf Abd El Hakeem Elkomy,

2Samar Saber Ibrahim Mohamed and

3*Mahmmoud Elsayed Mahmmoud Elmehalawy

1Pharmacology Department, Faculty of Veterinary Medicine Benha University, Egypt.

2Forensic Medicine Deparment, Benha University, Egypt.

3Glopal Human Health Department,

Merck Sharp & Dhome, Fifth Settlement, Egypt.

ABSTRACT

Ceftriaxone is a broad-spectrum semisynthetic cephalosporin antibiotic

that causes partial damage in the liver manifested by transient elevation

in some biochemical parameters. In this study, our aim was to

investigate the use of Ginseng in prevention of the hepatotoxic effect

and biochemical changes induced by ceftriaxone in rats. Rats were

divided into Five groups (control, ceftriaxone 180 mg/kg, Ginseng +

ceftriaxone 180 mg/kg, ceftriaxone 360 mg/kg, and Ginseng +

ceftriaxone 360 mg/kg). Ceftriaxone was injected intraperitone- ally,

and Ginseng was given orally daily for four consecutive weeks. Then

liver functions (serums AST, ALT, ALP, direct bilirubin, and total

protein) were assessed. Histopathological examination was performed.

Treatment of animals with ceftriaxone caused elevated activities of serum alanine

aminotransferase (ALT) and aspartate aminotransferase (AST) as well as total bilirubin level.

These elevations in liver enzymes were decreased by combination ceftriaxone with Ginseng.

In addition, ceftriaxone caused a significant increase in malondialdehyde (MDA) and nitric

oxide (NO) content but significant decrease in glutathione (GSH) content. Combination of

GINSENG and ceftriaxone resulted in a significant decrease in MDA, NO content and

significantly elevated GSH content. It could be concluded that GINSENG acts as an effective

hepatoprotective agent against liver dysfunction caused by ceftriaxone, and this effect might

be related to its antioxidant properties. Hepatic functions should be monitored, and the dose

should be adjusted during ceftriaxone therapy.

WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES

SJIF Impact Factor 7.632

Volume 9, Issue 3, 285-296 Research Article ISSN 2278 – 4357

*Corresponding Author

Mahmmoud Elsayed

Mahmmoud Elmehalawy

Glopal Human Health

Department, Merck Sharp &

Dhome, Fifth Settlement,

Egypt.

Article Received on

14 Jan. 2020,

Revised on 03 Feb. 2020,

Accepted on 23 Feb. 2020

DOI: 10.20959/wjpps20203-15680


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KEYWORDS: Ceftriaxone; GINSENG Hepatotoxicity; Liver, Kidney Protective Role.

1. INTRODUCTION

Liver injury caused by drugs ranges from mild biochemical abnormalities to acute and

chronic liver failure. The majority of adverse liver reactions is idiosyncratic, and occurs in

most instances 5–90 days after the causative medication was last taken.[1]

Some antibiotics are considered a common cause of drug- induced liver injury.[2]

Hepatotoxicity that occurs is usually asymptomatic, transient and associated with hepatic

impairment.3 Ceftriaxone is a broad-spectrum parenteral cephalosporin with potent activity

against gram-positive and gram-negative bacteria.[4]

It widely used, because of its pro- longed

terminal half-life that allows its prescription as a single dose per day.[5]

Hepatotoxicity caused

by ceftriaxone appears after 9–11 days.[6,7]

Previous studies have reported high aspartate

aminotransferase (ALT) and alanine aminotransferase (AST) activities with the

administration of ceftriaxone.[8,9]

Ceftriaxone causes partial damage in the liver as a result of transient elevation in some

biochemical parameters such as AST, ALT, total bilirubin, cholesterol, triglyceride (TG) and

Low-density lipoprotein (LDL) as well as transient decrease in albumin and High-density

lipoprotein (HDL) concentrations.[10]

Liver diseases are amongst the most serious health problems in the world today and

hepatocellular carcinoma is one of the world's deadliest cancers. The aim of the current study

was to evaluate the protective effect of sider honey and/or Korean ginseng extract (KGE)

against carbon tetrachloride (CCl4)-induced hepato-nephrotoxicity in rat. Eighty male

Sprague-Dawley (SD) rats were allocated into different groups and over a 4-week period,

they orally received honey and/or KGE or were treated either with CCl4 alone (100mg/kg

b.w) or with CCl4 after a pretreatment period with honey, KGE or a combination of both.

Clinical, clinico-pathological and histopathological evaluations were done and CCl4-treated

groups were compared with rats receiving no treatment and with rats given honey, KGE or a

combination of these substances. The results indicated that oral administration of CCl4

induced severe hepatic and kidney injury associated with oxidative stress. The combined

treatment with CCl4 plus honey and/or KGE resulted in a significant improvement in all

evaluated parameters. This improvement was prominent in the group receiving CCl4 after

combined pretreatment with honey and KGE. Animals receiving honey and/or KGE (without


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CCl4-treatment) were comparable to the control untreated group. It could be concluded that

honey and KGE protect SD rats against the severe CCl4-induced hepatic and renal toxic

effects. Our results suggest that the protective activity of honey and KGE may have been

related to their antioxidant properties.

2. MATERIALS AND METHODS

2.1. Animals

The present study was carried out on a total number of 35 white Albino male rats weighting

185-210 gm. Rats were obtained from Center of Laboratory Animal, Faculty of Veterinary

Medicine, Benha University, Egypt. They acclimatized for one week prior to the experiment.

All rats received standard laboratory balanced commercial diet and water ad libitum.

Drugs

Ceftriaxone (Mesporin®): Injectable solution, Each vial contains Mesporin sodium

equivalent to 250 mg, 500 mg, 1 gram or 2 grams of Mesporin activity. Mesporin for

Injection, USP contains approximately 83 mg (3.6 mEq) of sodium per gram of Mesporin

activity for EIPICO Egypt.

Gensing (Gensing®

): An oral Gelatin Capsule, hydroalcoholic extract of Ginseng root

(Panax Ginseng), containing 4% ginsenoside - 100 mg; It was purchased from Pharco group.

2.2. Experimental design

The rats were divided randomly into five experimental groups, each consisting of Seven rats,

that were treated as follows: group 1 received vehicle and served as a control, group 2

ceftriaxone (180 mg/kg), group 3 received combined oral doses of GINSENG 20 mg/kg and

ceftriaxone 180 mg/kg, group 4 received ceftriaxone (360 mg/kg) and finally group 5

received combined oral doses of GINSENG 20 mg/kg and ceftriaxone (360 mg/kg). Ceftri-

axone was i.m. injected while GINSENG was orally administered for 3 weeks.

At the end of the experiment blood samples were collected from the retro-orbital plexus and

used for serum separation. All the rats were sacrificed by decapitation and the livers of rats

were immediately dissected out. Part of the liver tissues was homogenized in ice-cold 0.9%

w/v saline using a homog- enizer to obtain 20% homogenate. Aliquots of the liver

homogenate were stored at 4 °C prior to biochemical analysis. The other part of the liver

was preserved in 10% formalin solution for histopathological examination.


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2.3. Determination of biochemical parameters

Hepatic enzymes in the serum such as AST and ALT were used as biochemical markers for

hepatotoxicity and assayed by the method of Reitman and Frankel.[17]

Serum alkaline

phosphatase (ALP) was determined according to the method of Belfield and Goldberg[18]

using colorimetric kit obtained from Diamond Co., Egypt. Total serum bilirubin was

determined spectrophotometrically according to the method of Walter and Gerade.[19]

2.4. Preparation of sections for histopathological examination

Formalin (10%): from Middle East Company, Cairo, Egypt.

Hematoxylin and Eosin (H&E) stain :from Middle East Company, Cairo, Egypt.

2.5. Statistical analysis

Statistical analysis was performed using SPSS (Version 20.0; SPSS Inc., Chicago, IL, USA).

The significant differences between groups were evaluated by one way ANOVA using

Duncan test as a post hoc. Results are expressed as mean ± SEM. P<0.05 was considered

significant.

Data analysis was accomplished using the software program graphpad prism (version 5).

3. RESULTS

GROUP OF RATS KEPT AS CONTROL

LIVER

There was no histopathological alteration and the normal histological structure of the central

vein and surrounding hepatocytes in the parenchyma were recorded in (Fig.1).

KIDNEY

There was no histopathological alteration and the normal histological structure of the

glomeruli and tubules at the cortex were recorded in (Fig.2).

GROUP OF RATS ADMINISTRATED THERAPEUTIC DOSE OF THE

CEFTRIAXONE

LIVER

Dilatation and congestion were detected in the portal vein (Fig.4).

KIDNEY

The cortical area showed congestion in the blood vessels and degeneration in the lining


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epithelial cells of some tubules (Fig.5).

GROUP OF RATS ADMINISTRATED THE DOUBLE THREAPEUTIC DOSE OF

THE CEFTRIAXONE

LIVER

The central and portal veins were dilated and congested (Fig.7) associated with inflammatory

cells infiltration in the portal area surrounding the bile ducts and congested portal vein

(Fig.8). There were dilatation in the hepatic sinusoids and microvacuolar degeneration in the

cytoplasm of some hepatocytes (Fig.9.)

KIDNEY

Perivascular inflammatory cells infiltration with oedema were detected surrounding the

congested cortical blood vessels (Fig.10). The corticomedullary junction showed focal

haemorrhages in between the tubules (Fig.11).

GROUP OF RATS ADMINISTRATED GINSENG AND THERAPEUTIC DOSE OF

THE CEFTRIAXONE

LIVER

Few inflammatory cells infiltration was detected in the portal area while the hepatocytes

surrounding the dilated central vein showed microvacuolar degeneration in the cytoplasm

(Fig.13).

KIDNEY

The cortex showed focal inflammatory cells infiltration in between the tubules (Fig.14)

associated with perivascular inflammatory cells infiltration surrounding the congested blood

vessels (Fig.15).

GROUP OF RATS ADMINISTRATED GINSENG AND DOUBLE DOSE OF THE

CEFTRIAXONE

LIVER

There was no histopathological alteration as recorded in (Fig.17).

KIDNEY

The cortical portion showed degeneration in the lining epithelial cells of some tubules

(Fig.18).


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4. DISCUSSION

The use of herbal medicines has increased considerably, because they are becoming a popular

alternative treatment in different countries. Plants constitute an important source of active

natural products with different biological properties. Various phytochemical components,

especially polyphenols (such as flavonoids, phyenyl propanoids, phenolic acids, tannins, etc)

are known to be responsible for the free radical scavenging and antioxidant activities of

plants. The got outcomes indicated that organization of ceftriaxone incited an assortment of

symptoms. These are spoken to by decrease of testicles, epididymis and adornment sex organ

weight, changes in sperm characters (lessening of sperm check and motility) and increment of

sperm variationsIn this manner, alert is required when utilizing enormous portions of

ceftriaxone because of its lethality to conceptive organs just as its expanding impact on liver

compound.Discover the Frequency of hepatotoxicity brought about by the wide range anti-

toxin mix amoxicillin-clavulanic corrosive (Co-amoxyclav) has been progressively perceived

and the component of this harmfulness stays vague. Then again, Ursodeoxycholic corrosive

(UDCA) has been recommended as effective cancer prevention agent treatment in different

liver ailments. Thusly, the present examination was intended to clarify the conceivable job of

oxidative worry in hepatotoxicity prompted by Co-amoxyclav and the putative defensive job

of UDCA in rodents. Impacts of amoxicillin (Amox; 50 mg/kg, orally, 21 d) or clavulanic

corrosive (Clav; 10 mg/kg, orally, 21 d) and their consolidated organization on the

biochemical liver parameters, decreased glutathione (GSH), lipid peroxidation estimated as

hepatic malondialdehyde (MDA) levels. Furthermore, myeloperoxidase (MPO) action and

receptive oxygen species (ROS) creation in liver homogenate were likewise assessed. Then

again, the defensive impacts of pretreatment with UDCA (20 mg/kg, orally, 21 d) on these

parameters were likewise assessed. Our outcomes show that pretreatment with UDCA

diminished the liver parameters that were upgraded by single or joined organization of Amox

or potentially Clav, for example, serum exercises of alanine aminotransferase (ALT),

aspartate aminotransferase (AST), soluble phosphatase (Snow capped mountain) and serum

bilirubin levels. In addition, pretreatment with UDCA standardized the GSH level and

hindered the height in hepatic MDA focus. The upgraded MPO action and ROS generation in

liver homogenate of rodents treated with Clav or Co-amoxyclav were additionally

standardized by UDCA pretreatment. Taking everything into account, the present information

recommend that UDCA goes about as compelling hepatoprotective operator against liver

brokenness brought about by Co-amoxyclav and this impact is identified with its cancer

prevention agent properties. State As of now, the utilization of β-lactamase inhibitors in


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explicit obstruction instrument of β-lactamase creating creatures in blend with β-lactams

filled in as exceptionally compelling and promising treatment. It recapture the

defenselessness profile of microscopic organisms to blend operator. The present examination

explores lethality profile of comparative routine, an intense and synergistic blend of third era

cephalosporins and β-lactamase inhibitor, Ceftriaxone-Tazobactam by directing rehashed

portion subchronic danger study on rodent (male and female). Three portion levels were

chosen for the investigation. Results of present examination proposed no critical changes in

physiological, biochemical just as hematological parameters. No mortality was seen in any of

the treatment gatherings. It was presumed that Ceftriaxone-Tazobactam mix applies no

danger and is protected on long haul treatmen utilization.

Figure 1: A There was no histopathological alteration and the normal histological

structure of the central vein and surrounding hepatocytes in the parenchyma were

recorded (H&E, ·100).

Figure 2 There was no histopathological alteration and the normal histological structure

of the glomeruli and tubules at the cortex were recorded (H&E, 100).


292


Figure 4: Dilatation and congestion were detected in the portal vein (H&E, ·50).

Figure 5: The cortical area showed congestion in the blood vessels and degeneration in

the lining epithelial cells of some tubules.

The central and portal veins were dilated and congested (Fig.7) associated with

inflammatory cells infiltration in the portal area surrounding the bile ducts and

congested portal vein (Fig.8). There were dilatation in the hepatic sinusoids and

microvacuolar degeneration in the cytoplasm of some hepatocytes (Fig.9).


293


5. CONCLUSION

In conclusion; the results of the present study demonstrate that GINSENG has a

hepatoprotective effect against liver injury caused by ceftriaxone owing to its antioxidant and

immuno- modulatory properties. Further, clinical studies are required to confirm this effect.

6. Conflict of interest

The authors declare that there is no conflict of interest.

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