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ORIGINAL ARTICLE
Year : 2013  |  Volume : 3  |  Issue : 2  |  Page : 114-120

Effect of Vimliv on lipid profile and histopathology in ethanol-induced hepatotoxicity in albino Wistar rats


Department of Biochemistry, Muthayammal College of Arts and Science, Rasipuram, Namakkal Dist., Tamil Nadu, India

Date of Submission18-Jul-2012
Date of Acceptance10-Sep-2012
Date of Web Publication3-Jun-2013

Correspondence Address:
Murugan Rajadurai
Department of Biochemistry, Muthayammal College of Arts and Science, Rasipuram - 637 408, Namakkal Dist., Tamil Nadu
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/2231-0738.112831

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   Abstract 

Background: Liver disease is considered to be a serious health problem, as the liver is an important organ for the detoxification and deposition of endogenous and exogenous substances. Objective: The present study is carried out to investigate the effect of Vimliv in ethanol-induced hepatotoxicity. Materials and Methods: Ethanol (3 g/kg) was dissolved in water and injected intragastrically, for a period of 35 days. Vimliv was dissolved in carboxy methyl cellulose and administered to rats at doses of 25 and 50 mg/kg for a period of 35 days as coadministration. Results: Administration of Vimliv resulted in significant reduction of blood glucose, serum urea, creatinine, lipid profile and increase in albumin. Administration of Vimliv also shows better pathological effects on liver and kidney. Conclusion: Therefore, this study suggests that Vimliv has the hepatoprotective effect and consequently may alleviate liver and renal damage associated with ethanol-induced hepatotoxicity in rats.

Keywords: Ethanol, hepatotoxicity, histopatholgy, lipid profile, vimliv


How to cite this article:
Samundeeswari N, Rajadurai M, Ganapathy P, Shairibha SR. Effect of Vimliv on lipid profile and histopathology in ethanol-induced hepatotoxicity in albino Wistar rats. Int J Nutr Pharmacol Neurol Dis 2013;3:114-20

How to cite this URL:
Samundeeswari N, Rajadurai M, Ganapathy P, Shairibha SR. Effect of Vimliv on lipid profile and histopathology in ethanol-induced hepatotoxicity in albino Wistar rats. Int J Nutr Pharmacol Neurol Dis [serial online] 2013 [cited 2019 Jun 19];3:114-20. Available from: http://www.ijnpnd.com/text.asp?2013/3/2/114/112831


   Introduction Top


Liver disease is considered to be a serious health problem, as the liver is an important organ for the detoxification and deposition of endogenous and exogenous substance. [1] Among liver diseases, alcohol-induced liver injury is one of the world's major health problems and excessive alcohol consumption caused several fetal diseases, which are evidenced by considerable experimental and clinical studies. [2],[3]

High alcohol consumption results in critical problems in the body including alcohol liver diseases (ALD). [4] Many pathways are thought to be involved in ALD, including oxidative stress and mitochondrial damage. [5] Ethanol is metabolized to acetaldehyde in the body by enzyme catalysis; acetaldehyde is further oxidized to acetate and then changed into carbon dioxide via the citric acid cycle. [6] Ethanol also affects the immune system and alters cytokine production, in turn increasing the levels of hepatic triglycerides and lipid peroxidation and decreasing hepatic glutathione (GSH) content. GSH acts as a free radical scavenger and a regenerator of α-tocopherol, and plays a significant role in sustaining protein sulfhydryl groups. [7],[8]

Herbal remedies are rapidly gaining popularity throughout the world because of nil effect of conventional medicines. Herbal preparations are considered harmless because of their natural origin. [9] However, several herbal medicines are also reported to have hepatotoxic effects. [10],[11] In view of wide application of herbal medicines on health, a study was conducted on the safety profile of a popular herbal preparation, which is in clinical use in Ayurveda as tonic, aphrodisiac, demulcent, lactogogue, purgative, and cholagogue [12] in various formulations. The present study was performed to assess the hepatoprotective effects of Vimliv on ethanol-induced rats, and the possible biochemical and morphologic changes in the liver and kidney. Vimliv fortified is a rationale combination of hepatoprotective herbs mentioned in Ayurvedic scriptures for the treatment of liver ailments, the composition of Vimliv presented in [Table 1].
Table 1: Composition of Vimliv

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   Materials and Methods Top


Experimental animals

All the experiments were done with female albino Wistar rats weighing 140-150 g, obtained from Venkateswara Enterprises, Bangalore were used in this study. They were housed in polypropylene cages (47 × 34 × 20 cm) lined with husk, renewed every 24 hours under a 12:12 hour light/dark cycle at around 22°C and had free access to water and food. The rats were fed on a standard pellet diet (Pranav Agro Industries Ltd., Maharashtra, India). The pellet diet consisted of 22.02% crude protein, 4.25% crude oil, 3.02% crude fiber, 7.5% ash, 1.38% sand silica, 0.8% calcium, 0.6% phosphorus, 2.46% glucose, 1.8% vitamins, and 56.17% nitrogen-free extract (carbohydrates). The diet provided metabolizable energy of 3,600 kcal. The experiment was carried out in accordance with the guidelines of the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), New Delhi, India.

Drugs and chemicals

Vimliv was purchased from local market, Salem, Tamil Nadu, India. Ethanol was purchased from Changshu Yangyuan Chemicals Pvt. Ltd., China. Chloroform, heptane, methanol, triethanolamine was purchased from Sigma Chemical Company, St. Louis, MO, USA. All other chemicals used in the study were of analytical grade.

Experimental induction of hepatotoxicity

Ethanol (3 gm/kg) was dissolved in water and injected intragastrically, for a period of 35 days. [13]

Experimental design

Vimliv tablet was made to powder with the help of mortar and pestle. The Vimliv powder administered at concentrations of 25 and 50 mg/kg dissolved in carboxy methyl cellulose (CMC) was given to rats through intragastric intubations for a period of 35 days.

In the experiment, a total of 30 rats (18 toxicity surviving rats, 12 control rats) were used in the study. The rats were divided into five groups of six rats each.

Group 1: Normal control rats
Group 2: Normal rats treated with Vimliv (50 mg/kg)
Group 3: Ethanol control rats
Group 4: Vimliv treated (25 mg/kg) + Ethanol
Group 5: Vimliv treated (50 mg/kg) + Ethanol

After the last treatment, all the rats were killed by cervical decapitation. Blood was collected for the estimation of blood glucose. Serum and plasma were separated from blood after centrifugation. The liver and kidney tissues were excised immediately from the rats, and washed off blood with ice-cold physiological saline. Known weight of the liver was homogenized in an appropriate buffer solution. The homogenates were centrifuged and the supernatants were used for the estimation of various biochemical parameters.

Biochemical estimation

Glucose was estimated by a Trinder [14] method, urea was estimated by the Chaney and Marbach [15] method, creatinine was estimated by the Henry [16] method and triglycerides (TG) levels were estimated by the Schettler and Nussel [17] method using a commercial kit from Agappe Diagnostics. [18]

Lipids were extracted from serum and liver by the method of Folch et al.,[19] using chloroform: methanol mixture (2:1 v/v). Total cholesterol (TC) was estimated by the method of Zlatkis et al. [20] Free fatty acids (FFA) levels were estimated by the method of Falholt et al. [21] Phospholipids (PL) levels were estimated by the method of Zilversmit and Davis. [22]

Histopathology

Tissues (liver and kidney) were obtained from all the experimental groups and washed immediately with saline and then fixed in a 10% buffered neutral formalin solution. After fixation, the tissues were processed by embedding in paraffin. Then, the tissues were sectioned and stained with hematoxylin and eosin (H and E) and examined under a high power microscope (400×) and photomicrographs were taken.

Statistical analysis

Statistical analysis was performed by one-way analysis of variance (ANOVA) followed by Duncan's multiple range test (DMRT) using a statistical package for the Social Sciences (SPSS) software package version 9.05. P values <0.05 were considered significant.


   Results Top


Effect of Vimliv on biochemical parameters

[Table 2] represents the levels of glucose, urea, creatinine, and A/G ratio in normal and ethanol-induced rats. Rats induced with ethanol exhibited a significant increase in the levels of these biochemical parameters with subsequent decrease in A/G ratio. Rats treated with Vimliv (25 and 50 mg/kg) for 35 days significantly minimized the alterations in the levels of these biochemical parameters.
Table 2: Effect of Vimliv on biochemical parameters

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Effect of Vimliv on the levels of lipids

[Figure 1], [Figure 2], [Figure 3], [Figure 4] and [Figure 5] shows the role of Vimliv on the levels of total cholesterol (TC), triglycerides (TG), free fatty acids (FFA), and phospholipids (PL) in serum and liver of normal and ethanol-administered rats. Ethanol control rats showed a significant increase in these lipid levels in serum and the liver. Oral treatment with Vimliv to ethanol-induced rats significantly decreased the levels of these lipids in serum and the liver.
Figure 1: Effect of Vimliv on the levels of serum total cholesterol and phospholipids (PL) in normal and ethanol-induced hepatotoxicity in rats Each value is mean ± S.D. for six rats in each group. Values not sharing a common superscript (a-d) differ signifi cantly from each other (P<0.05, DMRT)

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Figure 2: Effect of Vimliv on the levels of serum triglycerides (TG) and free fatty acids (FFA) in normal and ethanol-induced hepatotoxicity in rats

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Figure 3: Effect of Vimliv on the levels of liver free fatty acids (FFA) in normal and ethanol-induced hepatotoxicity in rats

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Figure 4: Effect of Vimliv on the levels of liver total cholesterol and triglycerides (TG) in normal and ethanol-induced hepatotoxicity in rats

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Figure 5: Effect of Vimliv on the levels of liver phospholipids (PL) in normal and ethanol-induced hepatotoxicity in rats

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Effect of Vimliv on histopathological examination

Liver histology

The effect of Vimliv on histopathology of liver is shown in [Figure 6], [Figure 7], [Figure 8], [Figure 9] and [Figure 10]. Normal control rats showed central veins with peripheral hepatic triads, showing normal architecture of the liver [Figure 6]. Ethanol-administered rats showed necrosis of the hepatocyte, mild bile duct hyperplasis, congestion with degeneration, and infiltration of mononuclear cells [Figure 8]. Rats treated with Vimliv (25 mg/kg) to ethanol-administered rats showed hydropic degeneration of hepatocytes and mild tubular epithelial cell degeneration and mononuclear cell infiltration [Figure 9]. Rats treated with Vimliv (50 mg/kg) to ethanol-administered rats showed mild tubular epithelial cell degeneration with necrosis [Figure 10]. Normal rats treated with Vimliv (50 mg/ kg) showed normal architecture of the hepatocytes without any pathological alterations [Figure 7].
Figure 6: Normal control rats showed normal architecture of the hepatocytes

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Figure 7: Normal rats treated with Vimliv (50 mg/kg) showed the normal architecture of the hepatocytes

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Figure 8: Ethanol control rats showed necrosis of the hepatocytes, congestion with degeneration and infi ltration of mononuclear cells

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Figure 9: Rats treated with Ethanol + Vimliv ( 25 mg/kg) showed hydropic degeneration of hepatocytes, with mild bileduct hyperplasia

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Figure 10: Rats treated with Ethanol + Vimliv ( 50 mg/ kg) showed mild tubular epithelial cells degeneration with necrosis

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Renal histology

The effect of Vimliv on histopathology of kidney is shown in [Figure 11], [Figure 12], [Figure 13], [Figure 14] and [Figure 15]. Normal control rats showed normal architecture of the renal tissue [Figure 11]. Ethanol-administered rats showed tubular epithelial cell degeneration with mononuclear cell infiltration with edema and necrosis [Figure 13]. Ethanol-administered rats treated with Vimliv (25 mg/kg) showed mild degeneration of epithelial cells with mild necrosis [Figure 14]. Ethanol-administered rats treated with Vimliv (50 mg/kg) showed mild necrosis and little degeneration of epithelial cells with near normal architecture of the kidney [Figure 15]. Normal rats treated with Vimliv (50 mg/kg) showed normal architecture of the kidney [Figure 12].
Figure 11: Normal control rats showed the normal architecture of the renal tissue

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Figure 12: Normal rats treated with vimliv (50mg/kg) showed the normal architecture of the kidney with very mild mononuclear cell infiltration

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Figure 13: Ethanol control rats showed tubular epithelial degeneration with mononuclear cell infiltration with oedema and necrosis

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Figure 14: Rats treated with ethanol + Vimliv (25 mg/ kg) showed mild degeneration of epithelial cells with mild necrosis

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Figure 15: Rats treated with ethanol + Vimliv ( 50 mg/kg) showed mild necrosis and a few degeneration of epithelial cells with near normal architecture

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   Discussion Top


The levels of glucose, urea, and creatinine were significantly increased with a subsequent decrease in the levels of A/G ratio in ethanol control rats. The elevated levels of glucose could be due to the toxic effect of ethanol on pancreas. The other biochemical parameters like urea, creatinine also increased, which could be due to the toxic effect of ethanol on hepatocytes. Total protein levels can be deemed as a useful index of the severity of cellular dysfunction in chronic liver diseases. [23] The levels of total proteins and A/G ratio were significantly decreased in ethanol control rats, may be associated with the decreased in the number of hepatocytes, which in turn, may result into the decreased hepatic capacity to synthesize protein.

Treatment with Vimliv at doses of 25 and 50 mg/ kg for a period of 35 days significantly increased the level of A/G ratio and decreased the levels of glucose, urea, and creatinine in ethanol-administered rats. This could be due to the protective effect of Vimliv on the hepatocytes, which minimized the destruction of hepatocytes and declined the permeability of liver cells.

Ethanol is known to have a profound effect on the metabolism of lipids and lipoproteins. Accumulation of lipids in the hepatocytes is the most striking initial manifestation of alcohol-induced liver injury. [24] The concentration of serum and tissue lipids such as free fatty acid (FFA), triacylglycerols (TG), total cholesterol (TC), and phospholipids (PL), is increased, in response to ethanol administration. Serum levels of lipids are principally a balance of production and deprivation by the liver. The observed increase in cholesterol level increases the membrane fluidity, disturbs its permeability, and alters the viscosity and also the internal chemical composition of membranes. [25] The role of high levels of cholesterol and TG in the early steps of atherosclerosis is well established. Increased levels of FFA may be due to lipid breakdown, which can increase the synthesis of other major lipids and activate NADPH- or NADH-dependent microsomal peroxidation. [26]

Phospholipids are basic machinery of cell membranes, mainly acting as regulators of membrane-bound enzymes and in membrane transport processes, and thus they are important in determining the pathology of various diseases. The increased PL content in the ethanol control group may be due to the elevated levels of FFA and cholesterol, which can promote the production of PL. [27]

Vimliv administration to ethanol-induced rats significantly decreased the levels of lipids in serum and liver. These effects may be related to the antioxidant and hypolipidemic effects of Vimliv. The decrease in FFA content, in Vimliv treated rats, may lead to the decreased synthesis of TG, cholesterol, PL, and extent of lipid peroxidation. The plant extracts and its active constituents may play a major role in the lipid lowering properties of Vimliv.

The effect of Vimliv on histopathology of liver normal control rats showed central veins with peripheral hepatic triads, showing normal architecture of the liver. Ethanol-administered rats showed necrosis of the hepatocyte, mild bile duct hyperplasis, congestion with degeneration and infiltration of mononuclear cells. Rats treated with Vimliv to ethanol-administered rats showed hydropic degeneration of hepatocytes and mild tubular epithelial cell degeneration and mononuclear cell infiltration.

The effect of Vimliv on histopathology of kidney in normal control rats showed normal architecture of the renal tissues. Ethanol-administered rats showed tubular epithelial cell degeneration with mononuclear cell infiltration with edema and necrosis. Rats treated with Vimliv to ethanol administered rats showed mild degeneration of epithelial cells with mild necrosis, with near normal architecture.


   Conclusion Top


The levels of lipids such as TC, TG, FFA, and PL were significantly increased in serum and liver of ethanol-induced rats. Vimliv treatment decreased these lipids levels to near normal. Histopathological studies also revealed that the protective effect of ethanol-induced rat liver and kidney. In conclusion, our observations suggested that treatment with Vimliv exhibited hepatoprotective effect against ethanol-induced hepatic damage in albino rats.

 
   References Top

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    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6], [Figure 7], [Figure 8], [Figure 9], [Figure 10], [Figure 11], [Figure 12], [Figure 13], [Figure 14], [Figure 15]
 
 
    Tables

  [Table 1], [Table 2]


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