Pre-treatment influences on SGPT levels and liver damage after inhalation of CCl4

Ten groups of male albino rats were subjected to different schedules of phenobarbitone treatment and/or starvation prior to a single 4 hr exposure to 1150 ppm CCl₄. By these treatments a gradual increase in liver cytochrome P-450 was achieved prior to exposure with a fourfold difference between the lowest and highest values. Rats were killed 20 hrs after exposure and it has been found that neither SGPT activity nor microscopic liver damage correlated with the pre-exposure values of P-450. The results seem to suggest that factors in addition to the level of cytochrome P-450 appear to determine SGPT levels and liver damage 20 hrs after exposure to CCl₄.     

Plasma amino acid levels after carbon tetrachloride induced acute liver damage. A dose-response and time-response study in rats

Summary The aims of the present study were to assess the changes of individual plasma amino acid levels in relation (1) to the severity of liver damage and (2) to the process of liver recovery. Acute liver injury was induced by an intragastric administration of CCl₄ diluted in olive oil in doses of 2, 4 and /or 6 g of CCl₄ per kg b.w. The control rats received olive oil only. Animals were sacrificed at 16, 24, 48 and 96 hours after treatment. The severity of liver injury was assessed by histological examination, by changes in ALT and AST in the blood plasma and by changes in liver weight. Statistical analysis was carried by ANOVA, p < 0.05 was considered significant. The Spearman rank correlation coefficient was used as a measure of the degree of linear relationship between variable and dose. In the period of the development of acute liver damage, i.e. at 16 and 24 hours after treatment, an increase in blood plasma amino acid levels and positive correlations with the dose of CCl₄ were observed for most individual amino acids. The only exception was arginine which decreased in a dose dependent manner. At a phase of liver recovery, i.e. at 48 and 96 hours after CCl₄ treatment, the concentrations of some individual amino acids decreased below the control values. The negative correlation with the dose of CCl₄ occurred for taurine and isoleucine (at 48 hours) and taurine, threonine, valine, methionine, isoleucine and leucine (at 96 hours).     

Perturbations in polyamines and related enzymes following chlordecone-potentiated bromotrichloromethane hepatotoxicity

The mechanism by which chlordecone (CD) amplifies the hepatotoxicity of halomethanes such as CCl₄, CHCl₃, and BrCCl₃ has been a subject of intense study. Recent work has shown that suppression of hepatocellular regeneration leads to accelerated progression of liver injury leading to complete hepatic failure due to an unusual interaction between individually nontoxic low-dose combination of CD and CCl₄. Since polyamines are involved in cell division, their levels reflect the extent to which there is suppression of hepatocellular regeneration during CD and CCl₄ interaction. The present studies were designed to investigate the polyamine levels and associated enzymes in livers of rats treated with BrCCl₃ alone or CD and BrCCl₃ low-dose combination in order to confirm whether the sequence of events of hepatotoxicity is similar to that seen in CCl₄ toxicity or that seen during CD and CCl₄ interaction. The extent of liver toxicity in rats fed 10 ppm chlordecone (CD) for 15 days prior to the injection of a single low dose of BrCCl₃ (15 μL/kg body weight) or after exposure to a high dose of BrCCl₃ (80 μL/kg body weight) without CD pretreatment, was similar 6 and 24 hr later as assessed by plasma transaminase levels. There was also an increase in transaminase levels, in rats exposed to a single low dose of BrCCl₃ alone (15 μL/kg body weight) but this increase was far below the high-dose exposure alone or the combination treatment. Hepatic levels of ornithine decarboxylase, S-adeno-sylmethionine decarboxylase, N¹-acetylputrescine, N¹-acetylspermidine, putrescine, spermidine, and spermine at the end of 24 hr increased after exposure to a low dose of BrCCl₃ alone as compared to exposure to a high dose alone or the low-dose combination of CD and BrCCl₃. Liver spermidine N¹-acetyltrans-ferase was elevated at 2, 6, and 24 hr after exposure to a high dose of BrCCl₃ alone as compared to treatment with a low-dose combination of CD and BrCCl₃ suggesting decreased synthesis of this enzyme, in spite of a greater need as seen from liver transaminase levels. In general, it was observed that there is significant elevation in some polyamines and related enzymes during toxicity of a low dose of BrCCl₃ which seemed to stabilize within 24 hr. This was not observed with the other two groups of rats exposed either to BrCCl₃ high dose alone or the low-dose combination of CD and BrCCl₃. Results indicate that CD and BrCCl₃ low-dose combination treatment causes increased liver toxicity resulting in compromised polyamine metabolism which is coincidental with suppressed hepatocellular regeneration leading to accelerated progressive phase of liver injury culminating in complete hepatic failure. These findings point to the possibility that the mechanism of potenti-ation of BrCCl₃ hepatotoxicity by CD is similar to that seen for CD and CCl₄ interaction.     

Neuroprotection against CCl4 induced brain damage with crocin in Wistar rats

Owing to its lipophilic property, carbon tetrachloride (CCl₄) is rapidly absorbed by both the liver and brain. We investigated the protective effects of crocin against brain damage caused by CCl₄. Fifty rats were divided into five groups of ten: control, corn oil, crocin, CCl₄ and CCl₄ + crocin. CCl₄ administration decreased glutathione (GSH) and total antioxidant status (TAS) levels, and catalase (CAT) activity, while significant increases were observed in malondialdehyde (MDA) and total oxidant status (TOS) levels and superoxide dismutase (SOD) activity. The cerebral cortex nuclear lamina developed a spongy appearance, neuronal degeneration was observed in the hippocampus, and heterochromatic and pyknotic neurons with increased cytoplasmic eosinophilia were observed in the hippocampus after CCl₄ treatment. Because crocin exhibits strong antioxidant properties, crocin treatment increased GSH and TAS levels and CAT activities, and decreased MDA and TOS levels and SOD activity; significant improvements also were observed in histologic architecture. We found that crocin administration nearly eliminated CCl₄ induced brain damage by preventing oxidative stress.     

Role of IL-6 trans-signaling in CCl4 induced liver damage

Interleukin-6 (IL-6) plays an important role in liver regeneration and protection against liver damage. In addition to IL-6 classic signaling via membrane bound receptor (mIL-6R), IL-6 signaling can also be mediated by soluble IL-6R (sIL-6R) thereby activating cells that do not express membrane bound IL-6R. This process has been named trans-signaling. IL-6 trans-signaling has been demonstrated to operate during liver regeneration. We have developed methods to specifically block or mimic IL-6 trans-signaling. A soluble gp130 protein (sgp130Fc) exclusively inhibits IL-6 trans-signaling whereas an IL-6/sIL-6R fusion protein (Hyper-IL-6) mimics IL-6 trans-signaling. Using these tools we investigate the role of IL-6 trans-signaling in CCl₄ induced liver damage. Blockade of IL-6 trans-signaling during CCl₄ induced liver damage led to higher liver damage, although induction of Cyp4502E1 and thus bioactivation of CCl₄ was unchanged. Depletion of neutrophils resulted in reduced liver transaminase levels irrespective of IL-6 trans-signaling blockade. Furthermore, IL-6 trans-signaling was important for refilling of hepatocyte glycogen stores, which were depleted 24 h after CCl₄ treatment. We conclude that IL-6 trans-signaling via the soluble IL-6R is important for the physiologic response of the liver to CCl₄ induced chemical damage.     

Potential role of regucalcin as a specific biochemical marker of chronic liver injury with carbon tetrachloride administration in rats

The potential sensitivity of liver specific protein regucalcin as a biochemical marker of chronic liver injury with carbon tetrachloride (CCl₄) administration in rats was investigated. CCl₄ (10%; 1.0 ml/100 g body wt) was orally given 5 times at 3-day intervals to rats, and the animals were killed by bleeding at 3, 6, 18, and 30 days after the first administration of CCl₄. The body weight of rats was significantly lowered 3 and 6 days after CCl₄ administration as compared with that of control rats administered with corn oil, and then the weight was restored at 18 and 30 days. Serum glutamate-oxaloacetate transaminase (GOT) and glutamate-pyruvate transaminase (GPT) activities were significantly increased 3 days after the administration, while a significant increase in serum -glutamyltranspeptidase (-GTP) activity was seen at 3 and 6 days after the administration. Serum GOT, GPT, and -GTP activities were restored to control levels at 18 and 30 days after CCl₄ administration. Serum albumin, -fetoprotein, and ammonium levels were not changed by CCl₄ administration. Meanwhile, serum regucalcin concentration was markedly increased 3 and 6 days after CCl₄ administration, and a significant increase in serum regucalcin concentration was observed 18 and 30 days after the administration. Liver regucalcin mRNA and liver cytosolic regucalcin levels were significantly decreased 18 and 30 days after CCl₄ administration. Liver content of calcium, which intracellular calcium homeostasis is maintained, was significantly increased between 3 and 30 days after CCl₄ administration. Hepatic mitochondrial succinate dehydrogenase activity was significantly increased 30 days after the administration. The present study demonstrates that serum regucalcin has a potential sensitivity as a specific biochemical marker of chronic liver injury with CCl₄ administration in rats.     

The protective effect of vitamin D against carbon tetrachloride damage to the rat liver

We investigated the protective effect of vitamin D against liver damage caused by carbon tetrachloride (CCl₄). Twenty-four male rats were divided into four equal groups: G1, untreated controls; G2, administered CCl₄; G3, administered both CCl₄ and vitamin D for 10 weeks; G4, administered CCl₄ for 10 weeks and vitamin D for 12 weeks. At the end of experiment, intracardiac blood samples were taken and liver samples were removed. Hepatic damage due to CCl₄ was assessed using biochemistry and histopathology. Glutathione (GSH) levels decreased, while malondialdehyde (MDA) levels increased in liver tissues of G2. Alanine transaminase (ALT), alkaline phosphatase (ALP), and gamma-glutamyl-transaminase (GGT) levels increased, while albumin (ALB) levels decreased. Hepatocyte degeneration, lobular disorder, sinusoid dilation, focal necrotic areas, hyperemia, and glycogen loss were observed. Hepatic fibrosis was observed around portal areas and central veins. Bridging fibrous septa were formed between portal veins. By immunohistochemistry, both matrix metalloproteinase-9 (MMP-9) and desmin reactivity were increased. All aspects of liver damage were at least partially prevented in rats treated with vitamin D. Vitamin D appears to act as an antioxidant and anti-fibrotic to protect the rat liver against damage.     

Distinct roles of ecto-nucleoside triphosphate diphosphohydrolase-2 (NTPDase2) in liver regeneration and fibrosis

Ecto-nucleoside triphosphate diphosphohydrolases (E-NTPDases) are cell surface-located transmembrane ecto-enzymes of the CD39 superfamily which regulate inflammation and tissue repair by catalyzing the phosphohydrolysis of extracellular nucleotides and modulating purinergic signaling. In the liver, NTPDase2 is reportedly expressed on portal fibroblasts, but its functional role in regulating tissue regeneration and fibrosis is incompletely understood. Here, we studied the role of NTPDase2 in several models of liver injury using global knockout mice. Liver regeneration and severity of fibrosis were analyzed at different time points after exposure to carbon tetrachloride (CCl₄) or 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) or partial hepatectomy in C57BL/6 wild-type and globally NTPDase2-deficient (Entpd2 null) mice. After chronic CCl₄ intoxication, Entpd2 null mice exhibit significantly more severe liver fibrosis, as assessed by collagen content and histology. In contrast, deletion of NTPDase2 does not have a substantial effect on biliary-type fibrosis in the setting of DDC feeding. In injured livers, NTPDase2 expression extends from the portal areas to fibrotic septae in pan-lobular (CCl₄-induced) liver fibrosis; the same pattern was observed, albeit to a lesser extent in biliary-type (DDC-induced) fibrosis. Liver regeneration after partial hepatectomy is not substantively impaired in global Entpd2 null mice. NTPDase2 protects from liver fibrosis resulting from hepatocellular injury induced by CCl₄. In contrast, Entpd2 deletion does not significantly impact fibrosis secondary to DDC injury or liver regeneration after partial hepatectomy. Our observations highlight mechanisms relating to purinergic signaling in the liver and indicate possible therapeutic avenues and new cellular targets to test in the management of hepatic fibrosis.     

Changes in rat plasma-free fatty acid composition under oxidative stress induced by carbon tetrachloride: decrease of polyunsaturated fatty acids and increase of palmitoleic acid

Summary

In order to measure the changes in antioxidant levels and the composition of plasma-free fatty acids resulting from oxidative stress, male Fisher rats were given a twice weekly subcutaneous injection of a 50% solution of carbon tetrachloride (CCl₄) in corn oil for a period of 2 to 13 weeks. The dosage was 1.3 ml/kg of body weight. This treatment significantly suppressed the gain of body weight compared with control rats receiving the same dosage of corn oil. Liver weight of the two groups was similar while necrosis and hyperplasia of hepatocytes and liver fibrosis were observed in CCl₄ treated rats. Increased levels of plasma glutamate-oxalacetate transaminase and glutamate-pyruvate transaminase were indicative that hepatocyte necrosis was induced by CCl₄. Increased oxidative stress in CCl₄ treated rats was indicated by a significant decrease of liver ascorbate and a decrease in the plasma ratio of polyunsaturated fatty acids (PUFA) to total free fatty acids. Interestingly, a significant increase of palmitoleic acid was observed in CCl₄ treated rats, which may compensate for the loss of PUFA. The possibility of using the plasma composition of PUFA and palmitoleic acid as a marker of oxidative stress is discussed.     

Serum hormone levels following partial hepatectomy in the rat.

Serum levels of insulin, glucagon, growth hormone (somatotrophin) and thyroxine (TT₄) were measured by radioimmunoassay following both sham operation and 70% partial hepatectomy in the rat to evaluate changes in hormone levels during liver regeneration. An eleven fold increase in glucagon was observed (from 112 ± 10 pg/ml to 1500 ± 200 pg/ml) 6 hours following partial hepatectomy but not sham operation. In contrast, insulin levels remained unchanged compared to sham controls for up to 72 hr while growth hormone fell to low levels, 6 to 48 hr after partial hepatectomy. Both total thyroxine and free thyroxine levels also fell 24–72 hours after hepatectomy. These studies suggest that growth hormone, thyroxine and insulin are not primary stimulants of hepatic regeneration although the data suggests that glucagon may modify this growth process.     

Induction of liver fibrosis by CCl4 mediates pathological alterations in the spleen and lymph nodes: The potential therapeutic role of propolis

In an animal models, carbon tetrachloride (CCl₄) is a carcinogenic agent that causes liver fibrosis. The current study aims to investigate whether induction in liver-fibrosis by CCl₄ in the mouse model could promote the initiation of fibrosis in lymph node and spleen due to sustained increase of inflammatory signals and also aimed to clarify the protective therapeutic effects of propolis. The male mice (BALB/c) were categorized into three experimental sets and each group involved 15 mice. Control group falls into first group; group-II and group-III were injected with CCl₄ to induce liver-fibrosis and oral supplementation with propolis was provided in group-III for 4-weeks. A major improvement with hepatic collagen and α-smooth muscle actin (α-SMA) production was aligned with the activation of liver fibrosis from CCl₄. Mice treated with CCl₄ exhibited collagen deposition towards liver sections, pathological alterations in spleen and lymph node architectures, and a significantly increase the circulation of both T&B cells in secondary lymphoid organs. Mechanically, the secondary lymphoid organs treated with CCl₄ in mice exposed a positive growth in α-SMA and collagen expression, increased in proinflammatory cytokine levels and a significant increase in TGF-β, NO and ROS levels. A manifest intensification in the expression of Nrf2, COX-2, and eNOS and upregulation of ASK1 and P38 phosphorylation. Interestingly, addition of propolis-treated CCl₄ mice, substantially suppressed deposition of liver collagen, repealed inflammatory signals and resorted CCl₄-mediated alterations in signaling cascades, thereby repairing the architectures of the secondary lymphoid organs. Our findings revealed benefits of propolis against fibrotic complications and enhancing secondary lymphoid organ architecture.     

3-Alkynyl selenophene protects against carbon-tetrachloride-induced and 2-nitropropane-induced hepatic damage in rats

The aim of this study was to investigate the protective effect of 3-alkynyl selenophene (3-ASP) on acute liver injury induced by carbon tetrachloride (CCl₄) and 2-nitropropane (2-NP) in rats. On the first day of treatment, the animals received 3-ASP (25 mg/kg, p.o.). On the second day, the rats received CCl₄ (1 mg/kg, i.p.) or 2-NP (100 mg/kg, p.o.). Twenty-four hours after CCl₄ or 2-NP administration, the animals were euthanized, and their plasma and liver were removed for biochemical and histological analyses. The histological analysis revealed extensive injury in the liver of CCl₄-exposed and 2-NP-exposed rats, which was attenuated by 3-ASP. 3-ASP significantly attenuated (1) the increase in plasmatic aspartate and alanine aminotransferase activities and lipid peroxidation levels induced by CCl₄ and 2-NP; (2) the inhibition of δ-aminolevulinic dehydratase activity caused by 2-NP; and (3) the decrease in ascorbic acid (AA) levels and catalase (CAT) activity caused by CCl₄. AA levels and CAT activity remained unaltered in the liver of rats exposed to 2-NP. The protective effect of 3-ASP on acute liver injury induced by CCl₄ and 2-NP in rats was demonstrated.     

Changes in the insulin and glucagon receptors in the regenerating liver following intoxication with carbon tetrachloride.

The response of rat liver plasma membrane adenylate cyclase was studied from one to 14 days after a single dose of carbon tetrachloride (CCl₄). The response to glucagon was diminished to a greater extent than that of fluoride and was due to a deficiency in hormone binding. In contrast, insulin binding increased 300% over control; the change was due to increased number of binding sites. The “affinities” of receptors for either hormone were not altered. The tissue levels of adenosine 3′:5′ -monophosphate increased following CCl₄ poisoning reaching a peak precisely when the adenylate cyclase response to glucagon was at its lowest level. These studies present evidence that receptors for pancreatic hormones change differently when liver is damaged and during its regeneration following CCl₄ intoxication. The change in pattern is remarkably similar to changes reported previously in fetal liver development or following partial hepatectomy of adult rat.     

Colchicine alters lactate utilization in isolated hepatocytes of rats treated with CCl4 and ethanol

Lactic acidosis has been described in patients with liver disease. Hyperlactacidaemia results from an imbalance in lactate production versus lactate utilization. It is estimated that the liver utilizes approximately 30 percent of the total lactate produced in the body under basal conditions, primarily by gluconeogenesis. The gluconeogenesis from lactate 10 mM and lactacidaemia were determined in order to investigate the effects of CCl₄+ethanol administration in liver injury and, the possible effect of colchicine in our experimental fibrosis model. The tests were determined after 15, 30 or 45 days of treatment. The results indicate that the gluconeogenesis was significantly inhibited in both CCl₄+ethanol groups and CCl₄+ethanol+colchicine groups. By contrast, the lactacidaemia levels were much higher in the CCl₄+ethanol groups than the colchicine groups. Summarising, we have documented that hyperlactacidaemia is due to the inhibition of lactate utilization by the isolated hepatocytes in experimental cirrhosis, and that the improvement in lactacidaemia caused by colchicine is not primarily due to an increase in hepatic lactate utilization.     

Zonation of Nitrogen and Glucose Metabolism Gene Expression upon Acute Liver Damage in Mouse

Zonation of metabolic activities within specific structures and cell types is a phenomenon of liver organization and ensures complementarity of variant liver functions like protein production, glucose homeostasis and detoxification. To analyze damage and regeneration of liver tissue in response to a toxic agent, expression of liver specific enzymes was analyzed by in situ hybridization in mouse over a 6 days time course following carbon tetrachloride (CCl₄) injection. CCl₄ mixed with mineral oil was administered to BALB/c mice by intraperitoneal injection, and mice were sacrificed at different time points post injection. Changes in the expression of albumin (Alb), arginase (Arg1), glutaminase 2 (Gls2), Glutamine synthetase (Gs), glucose-6-phosphatase (G6pc), glycogen synthase 2 (Gys2), Glycerinaldehyd-3-phosphat-Dehydrogenase (Gapdh), Cytochrom p450 2E1 (Cyp2e1) and glucagon receptor (Gcgr) genes in the liver were studied by in situ hybridization and qPCR. We observed significant changes in gene expression of enzymes involved in nitrogen and glucose metabolism and their local distribution following CCl₄ injury. We also found that Cyp2e1, the primary metabolizing enzyme for CCl₄, was strongly expressed in the pericentral zone during recovery. Furthermore, cells in the damaged area displayed distinct gene expression profiles during the analyzed time course and showed complete recovery with strong albumin production 6 days after CCl₄ injection. Our results indicate that despite severe damage, liver cells in the damaged area do not simply die but instead display locally adjusted gene expression supporting damage response and recovery.     

Effects of indoleamine 2,3‐dioxygenases in carbon tetrachloride‐induced hepatitis model of rats

Indoleamine 2,3‐dioxygenase (IDO) converts tryptophan to l ‐kynurenine, and it is noted as a relevant molecule in promoting tolerance and suppressing adaptive immunity. In this study, to investigate the effects of IDO in carbon tetrachloride (CCl₄)–induced hepatitis model, the levels of IDO enzymic activities in the mock group, the control group and the 1‐methyl‐d ‐tryptophan (1‐MT)–treated group were confirmed by determination of l ‐kynurenine concentrations. Serum alanine aminotransferase levels in 1‐MT‐treated rats after CCl₄ injection significantly increased compared with those in mock and control groups. In CCl₄‐induced hepatitis models, tumour necrosis factor‐α (TNF‐α) is critical in the development of liver injury. The mRNA expression and secretion levels of TNF‐α in the liver from 1‐MT‐treated rats were more enhanced compared with those in the mock and the control groups. Moreover, the levels of cytokine and chemokine from mock, control group and 1‐MT‐treated rats after treated with CCl₄ were analyzed by ELISA, and the level of interleukin‐6 was found to increase in 1‐MT‐treated rats. It was concluded that the deficiency of IDO exacerbated liver injury in CCl₄‐induced hepatitis and its effect may be connected with TNF‐α and interleukin‐6. Copyright © 2012 John Wiley & Sons, Ltd.     

Immunoperoxidase labelling of alpha1-fetoprotein (AFP) in normal and regenerating livers of a low and a high AFP producing mouse strain

Summary Serum AFP concentrations in normal adult BALB/c/J and in normal adult C3H/He mice were in the order of 0.6 g/ml and 0.1 g/ml, respectively. In BALB mice, AFP was localized in the cytoplasm of differentiated mono- and binucleated hepatocytes in centrolobular and intermediate zones of normal adult liver. No cellular AFP could be detected in liver sections of normal adult C3H mice.CCl₄ intoxication was accompanied by increase of serum AFP levels. A maximum was reached on day 4. Afterwards, concentrations declined. In sera of BALB/c/J mice, AFP levels reached values 10-fold higher and more than in sera of C3H/He mice.From day one after CCl₄ intoxication, cellular AFP was detected in hepatocytes of portal and periportal areas including intermediate zones adjacent to the necrosis. The intensity of AFP staining reached a maximum between the days 3 and 4. Hepatocytes in front of the necrotic areas usually contained the strongest AFP reactions. In both mouse strains, cellular AFP pattern was comparable, but strongest immunoreactivity was observed in liver sections of BALB/c/J mice.Liver injury and subsequent regeneration occurred to the same extent in both studied strains. The much higher serum AFP levels and the stronger AFP immunolocalizations in BALB mice were thought not due to increased numbers of AFP producing and releasing cells during liver regeneration. Additional mechanisms must play a role in increased AFP synthesis per single cell. C3H/He was a low AFP-inducible and BALB/c/J was a high AFP-inducible mouse strain.Supported by the Deutsche Forschungsgemeinschaft (Ku 257/3) Bonn. Federal Republic of Germany     

Hepatoprotective effect of Origanum vulgare in Wistar rats against carbon tetrachloride-induced hepatotoxicity

The effect of an aqueous extract of Origanum vulgare (OV) leaves extract on CCl₄-induced hepatotoxicity was investigated in normal and hepatotoxic rats. To evaluate the hepatoprotective activity of OV, rats were divided into six groups: control group, O. vulgare group, carbon tetrachloride (CCl₄; 2 ml/kg body weight) group, and three treatment groups that received CCl₄ and OV at doses of 50, 100, 150 mg/kg body weight orally for 15 days. Alanine amino transferase (ALT), alkaline phosphatase (ALP), and aspartate amino transferase (AST) in serum, lipid peroxide (LPO), GST, CAT, SOD, GPx, GR, and GSH in liver tissue were estimated to assess liver function. CCl₄ administration led to pathological and biochemical evidence of liver injury as compared to controls. OV administration led to significant protection against CCl₄-induced hepatotoxicity in dose-dependent manner, maximum activity was found in CCl₄?+?OV3 (150 mg/kg body weight) groups and changes in the hepatocytes were confirmed through histopathological analysis of liver tissues. It was also associated with significantly lower serum ALT, ALP, and AST levels, higher GST, CAT, SOD, GPx, GR, and GSH level in liver tissue. The level of LPO also decreases significantly after the administration of OV leaves extract. The biochemical observations were supplemented with histopathological examination of rat liver sections. Thus, the study suggests O. vulgare showed protective activity against CCl₄-induced hepatotoxicity in Wistar rats and might be beneficial for the liver toxicity.