Carbon Tetrachloride Hepatotoxicity Research Articles

Ameliorative Effects of Pomegranate Molasses and Fig on Carbon Tetrachloride Hepatotoxicity in Rats

Ameliorative Effects of Pomegranate Molasses and Fig on Carbon Tetrachloride Hepatotoxicity in Rats

Modulation of miR-192/NF-κB/ TGF-β/ E-cadherin by thymoquinone protects against diethylnitrosamine /carbon tetrachloride hepatotoxicity.

Scientific efforts have been made for a better understanding of the pathogenesis of hepatocellular carcinoma (HCC). We investigated the possible role of miR-192/nuclear factor-κB (NF-κB)/transforming growth factor-β (TGF-β)/E-cadherin in hepatic tumorigenesis. We expected a modulatory impact of thymoquinone. Thirty adult male rats were assigned into 3 groups (n = 10); (1) Control group. Group (2): Experimental HCC induced by intraperitoneal injection of diethylnitrosamine (DENA) followed by carbon tetrachloride (CCl4). Group (3): Thymoquinone 20 mg kg-1/oral supplementation starting from the model induction to the end of the 8th week. The HCC (DENA-CCL4) model was confirmed by elevated serum levels of alpha-fetoprotein and transaminases (ALT, AST) and by histopathological examination which denoted marked cellular atypia and features of neoplasia. Suppressed hepatic miR-192 and E-cadherin expression were detected in the HCC (DENA-CCL4) group accompanied by elevated tumor necrosis factor (TNF-α), interleukin (IL6)/NF-κB & TGF-β1. Thymoquinone treatment protected the rat livers from hepatic tumorigenesis. Thymoquinone diminished (P < 0.001) alpha-fetoprotein and improved ALT, AST. It preserved hepatic miR-192 and normal E-cadherin expression. Thymoquinone-treated rats showed abrogated TNF-α, IL6/NF-κB/TGF-β. Thymoquinone increased cell apoptosis markers Bax/Bcl2 and diminished cellular atypia. Pearson's correlations revealed positive association between miR-192 expression and E-cadherin and Bax/Bcl2 as well, and it was negatively correlated to alpha-fetoprotein, NF-κB and TGF-β and the cellular atypia score. In conclusion, thymoquinone protected the liver tissues through preserving miR-192 and E-cadherin and aborting NF-κB & TGF-β signaling. The current results highlight a new role for thymoquinone in preventing hepatic tumorigenesis.

Effect of silymarin on blood coagulation profile and osmotic fragility in carbon tetrachloride induced hepatotoxicity in male Wistar rats

Reports about the impact of Carbon tetrachloride (CCl4) hepatotoxicity on coagulation profile have been inconsistent. Multiple investigators have however demonstrated the effectiveness of silymarin in the resolution of anomalies induced by CCl4, although the effect of silymarin on the impact of CCl4 hepatotoxicity, especially coagulation profile and osmotic fragility have not been investigated. The liver, the primary site for the secretion of coagulation proteins, can become impaired in CCl4 hepatotoxicity, and silymarin reportedly increases hepatic protein synthesis as part of its hepatoprotective mechanism. This study assessed the effect of silymarin on blood coagulation profile and erythrocyte osmotic fragility in CCl4 induced hepatotoxicity in rats. Twenty male Wistar rats were allocated into four groups (n = 5) at random, namely: Control, CCl4 given CCl4 (1 ml/kg) administered intraperitoneally twice a week, Silymarin (S) given silymarin (100 mg/kg/day) orally, and S+CCl4 given silymarin (100 mg/kg/day) orally and (1 ml/kg) CCl4 one hour after, intraperitoneally twice a week for a duration of four weeks. Results showed protraction of activated partial thromboplastin time and thrombin time, increased erythrocyte osmotic fragility, liver damage, dyslipidemia, oxidative stress and lipid peroxidation in rats given CCl4. Silymarin attenuated most of these effects as observed from comparison between CCl4 and S+CCl4 rats. The findings of this study suggests that pretreatment with silymarin attenuated disruption in coagulation profile and erythrocyte osmotic fragility in CCl4 induced hepatotoxicity in Wistar rats.

Ethanolic extract of Mucuna pruriens leaves ameliorates carbon tetrachloride and rifampicin-induced hepatotoxicity and nephrotoxicity in wistar albino rat

BackgroundMucuna pruriens (L.) has been used for the treatment of several ailments in folkloric medicine. The present study therefore investigates the hepatoprotective and nephroprotective potentials of its leaves extract with a view to providing a potent alternative in the management of liver and kidney diseases.MethodologyForty male albino rats were randomly placed into eight groups comprising five animals each. Animals in group I were administered with the distilled water, while groups II and VI were exposed to CCl4 and rifampicin respectively. Animals in groups III and IV were initially exposed CCl4 and treated with 50 and 100 mg/kg bw M. pruriens respectively. Similarly, groups VII and VIII animals were exposed to rifampicin and treated with 50 and 100 mg/kg bw M. pruriens respectively. Animals in group V were treated with 100 mg/kg bw silymarin by oral gavage after an initial exposure to CCl4. Selected biomarkers of liver and kidney damage were determined in the serum and organs homogenate. Liver and kidney slices of experimental animals were also stained for histopathological examination.ResultsExposure to CCl4 and rifampicin respectively resulted in marked distortion in lipid profile, inhibition of antioxidant enzymes and a surge in ALT, AST, ALP, urea, uric acid, bilirubin and creatine kinase. Treatment with M. pruriens extract reversed all deranged biochemical and histopathological parameters in a dose-dependent manner.ConclusionExtract of M. pruriens leaves restored deranged biochemical and histopathological parameters in the liver and kidney with similar potency to silymarin. Hence, leaf extract of M. pruriens is a potential hepatoprotective and nephroprotective agent that can be exploited in the management of liver and kidney diseases.

RNA-Seq analysis of the protection by Dendrobium nobile alkaloids against carbon tetrachloride hepatotoxicity in mice

ObjectiveDendrobium nobile is a genuine Chinese medicine. Dendrobium nobile Lindl. alkaloids (DNLA) protects against CCl4-induced acute liver injury. This study used RNA-Seq to explore the mechanisms. MethodsMice were pretreated with DNLA (10 and 20 mg/kg, po) for 7 days, and subsequently intoxicated with CCl4 (20 μL/kg, ip for 24 h). Liver RNA was extracted and subjected to RNA-Seq. The bioinformatics, including PCA, GO, KEGG, two-dimensional clustering, Ingenuity Pathways Analysis (IPA), and Illumina BaseSpace Correlation Engine (BSCE) were used to analyze the data. qPCR was performed on selected genes to verify RNA-Seq results. ResultsDNLA protection against CCl4 hepatotoxicity was confirmed by histopathology. PCA revealed the distinct gene expression patterns between the different treatment groups. GO showed that CCl4 induced the activation, adhesion and proliferation of immune cells. KEGG showed CCl4 induced oxidative stress, diseases and compromised adaptive responses. CCl4 induced differentially expressed genes (DEGs) were identified by DESeq2 with Padj < 0.05 and 2D-clustered with other groups. DNLA reverted CCl4-induced DEGs in a dose-dependent manner. qPCR analysis of S100 g, Sprr1, CCL3/7, Saa2/3, IL1rn, Cox7a2 and Rad15 confirmed RNA-Seq results. IPA showed that CCl4 treatment altered some signaling and metabolic pathways, which were ameliorated or returned to normal following DNLA treatment. The CCl4-activated mitochondrial oxidative phosphorylation was illustrated as an example. IPA Upstream Regulator Analysis further revealed the activated or inhibited molecules and chemicals that are responsible for CCl4-induced DEGs, and DNLA attenuated these changes. BSCE analysis verified that CCl4-induced DEGs were highly correlated with the GEO database of CCl4 hepatotoxicity in rodents, and DNLA dose-dependently attenuated such correlation. ConclusionRNA-Seq revealed CCl4-induced DEGs, disruption of canonical pathways, activation or inhibition of upstream regulators, which are highly correlated with database for CCl4 hepatotoxicity. All these changes were attenuated or returned to normal by DNLA, demonstrating the mechanisms for DNLA to protect against CCl4 hepatotoxicity.

Hepatoprotective Activity of the Traditional Multicomponent Formulation Gurgem-13 on Carbon Tetrachloride (CCL4) Induced Experimental Liver Injury in Rats

Objectives: The purpose of present study was intended to evaluate hepatoprotective effects of traditional multicomponent formulation Gurgum13 on carbon tetrachloride (CCI4) induced liver injury model in rats. Methods: Chronic toxic liver injury was induced experimentally by intraperitoneal injection (IP) of 1.0 ml/kg body weight of 10% carbon tetrachloride (CCI4) three times per week for duration of three months in rats. A total of five groups of animals with 15 rats in each were used for the experiment. Group 1 was as a normal control (intact). Group 2 served as a control group received CCI4 1.0 ml/kg IP three times a week for 3 months. Group 3, 4 and 5 were as the treatment groups received CCI4 1 ml/kg IP and administered traditional multicomponent formulation Gurgum-13 as a test drug (100 mg/kg) for 3 months daily. Milk thistle (50 mg/kg) and Lilicoaguliant (LLC) (100 mg/kg) were given daily for 3 months as a reference drug respectively. The activity of Gurgum-13 formulation and other comparative drugs were investigated with the dynamic study of liver marker enzymes, AST, ALT, ALP, bilirubin, and the antioxidant and blood coagulating effects of the remedy, which were studied in three months of experimentation. Gurgum-13 is one of the Mongolian traditional multicomponent formulation used in practice for a range of diseases including liver disorders. Results: Carbon tetrachloride significantly decreased the liver functions as assessed through an increase in blood serum and liver marker enzymes in control groups. Gurgum-13 treatment significantly prevented hepatotoxicity of carbon tetrachloride by reducing ALT, AST, ALP marker enzymes and bilirubin in blood serum and liver tissue in the three months period of chronic hepatitis (p&lt;.05) similar to reference drugs. The result of the study revealed that Gurgum-13 is significant hepatoprotective against liver injury from CCI4 induced hepatotoxicity by having antioxidant effect and decreasing lipid peroxidation in liver. Conclusion: Multicomponent traditional formulation Gurgum-13 demonstrated very good hepatoprotective effect against CCI4 induced liver injury in rats. Thus, Gurgum-13 can be an effective remedy for the treatment of chronic hepatitis and biliary tract disorders.

Hepcidin-orchestrated Hemogram and Iron Homeostatic Patterns in Two Models of Subchronic Hepatic injury

ObjectiveThis study was designed to evaluate hematological disorders and the orchestrating roles of hepcidin and IL-6 in rat models of thioacetamide (TAA) and carbon tetrachloride (CCl4) hepatotoxicity. MethodsRats were intraperitoneally injected with TAA (10 mg/100 g rat weight dissolved in isosaline) or CCl4 (100 μL/100 g rat weight diluted as 1:4 in corn oil) twice weekly for eight consecutive weeks to induce subchronic liver fibrosis. Blood and tissue samples were collected and analyzed. ResultsCCl4 but not TAA significantly decreased the RBCs, Hb, PCV, and MCV values with minimal alterations in other erythrocytic indices. Both hepatotoxins showed leukocytosis, granulocytosis, and thrombocytopenia. By the end of the experiment, the erythropoietin level increased in the CCl4 model. The serum iron, UIBC, TIBC, transferrin saturation%, and serum transferrin concentration values significantly decreased, whereas that of ferritin increased in the CCl4 model. TAA increased the iron parameters toward iron overload. RT-PCR analysis revealed increased expression of hepatic hepcidin and IL-6 mRNAs in the CCl4 model and suppressed hepcidin expression without significant effect on IL-6 in the TAA model. ConclusionThese data suggest differences driven by hepcidin and IL-6 expression between CCl4 and TAA liver fibrosis models and are of clinical importance for diagnosis and therapeutics of liver diseases.

Evaluation of Hepatotoxicity of Carbon Tetrachloride and Pharmacological Intervention by Vitamin E in Balb C Mice

Evaluation of Hepatotoxicity of Carbon Tetrachloride and Pharmacological Intervention by Vitamin E in Balb C Mice

Protective effect of pilose antler peptide on carbon tetrachloride-induced hepatotoxicity in mice

The present study was designed to investigate the beneficial effects of pilose antler peptide (PAP) against carbon tetrachloride (CCl4) hepatotoxicity in mice, and explore the underlying mechanisms. In our results, animals were treated with either CCl4 and/or PAP for six consecutive weeks. The levels of inflammatory cytokines in serum and liver, hepatotoxicity markers in serum and histopathological evaluation were determined. PAP (50, 100mg/kg) significantly inhibited the CCl4-incuded overproduction of inflammatory cytokines including tumor necrosis factor α (TNF-α), cytokines interleukin-1β (IL-1β), and interleukin-6 (IL-6) in serum and liver. Animals treated with PAP exhibited lower levels of alanine transaminase (ALT) and aspartate transaminase (AST), PAP administration alleviated CCl4 induced hepatic fibrosis as manifested in histopathological records. In fact, PAP prevented the activation of TLR/NF-κB pathway, by inhibiting the expression of TLR2, TLR4 MyD88, p-NF-κBp65 and p-IκBα. In addition, PAP also significantly reversed CCl4-induced alteration of TGF-β and p-samd-3 in liver tissue. In conclusion, PAP restored CCl4-induced hepatotoxicity via TLR/NF-κB and TGF-β/samd-3 pathways.

The immature rat as a potential model for chemical risks to children: Ontogeny of selected hepatic P450s

Concern about potential susceptibilities of infants and children to chemicals has led to the consideration of immature rodents as potential test surrogates. Maturation of some hepatic microsomal cytochrome P450s (CYPs), that participate in metabolic activation of organic solvents and polycyclic aromatic hydrocarbons (PAHs), may differ significantly between humans and rodents. The present investigation was undertaken to delineate the ontogeny of selected hepatic CYPs in male and female Sprague-Dawley (S-D) rats, and to contrast them with developmental profiles in humans. Microsomes were prepared from the liver of sexed and unsexed postnatal day (PND) 1–90 rats, and total CYP450 levels, as well as CYP1A1/2, CYP2E1 and CYP2B1/2 activities and protein, were quantified. CYP1A1/2 and CYP2E1 activity and expression rose rapidly after birth, peaked from PND 21–40/50, and declined substantially to adult values by PND 90. The same ontogenic profiles were manifested when the enzyme activities were expressed per entire liver or liver normalized to body weight. CYP1A1/2 and CYP2E1 activity and protein expression were well correlated. CYP2B1/2 activity peaked abruptly on PND 21 and declined irregularly to adult values. These patterns are in contrast to human CYP1A2 and CYP2E1, which are reported to progressively increase in liver during the first few months to years of life. The three CYP protein developmental profiles were largely gender independent in rats. The immature rat does not appear to be a suitable model for assessing risks posed to infants and children by chemicals metabolically activated by CYP2E1, based on the findings of greater carbon tetrachloride hepatotoxicity in preweanlings and weanlings than in adult animals. Additional studies are required to determine whether immature S-D rats may be used as an animal model for substrates of other CYPs, as total CYP450 levels in the liver progressively rose during maturation, similarly to humans.

Ameliorative effects of pomegranate on carbon tetrachloride hepatotoxicity in rats: A molecular and histopathological study.

The present study aimed to investigate the molecular mechanism underlying the hepatoprotective effects of pomegranate (POM) against oxidative stress in a rat model of carbon tetrachloride (CCl4)-induced liver damage. Injection of rats with CCl4 resulted in hepatic inflammation and lipid accumulation via the upregulation of interleukin (IL)‑6 and sterol regulatory element‑binding protein 1c (SREBP‑1c) mRNA expression. CCl4 induced downregulation of the anti‑inflammatory factors alpha 2‑macroglobulin (α‑2M) and IL‑10 in comparison with the POM treated group. In addition, CCl4 induced downregulation of superoxide dismutase (SOD), glutathione S‑transferase (GST) and catalase (CAT) expression. Conversely, prior administration of POM counteracted the deleterious alterations induced by CCl4. POM downregulated CCl4-induced IL‑6 upregulation, normalized the increase in SREBP‑1c expression, and prevented CCl4‑induced α‑2M downregulation. POM counteracted CCL4‑induced alterations via immunosuppressive, anti‑inflammatory and regenerative effects by upregulating transforming growth factor‑β1, HSP70 and IL-10 mRNA expression. In addition, POM increased reactive oxygen species scavenging activity by augmenting the antioxidant defense mechanism against CCl4 hepatotoxicity, as demonstrated by detecting SOD, CAT and GST expression. These results confirm that, at the molecular level, POM exerts hepatoprotective effects against CCl4‑induced oxidative stress and liver tissue damage.

The Effect of N-acetylcysteine on Pyrrolized Protein, Lipid Hydroperoxide and Thiol Levels in the Carbon Tetrachloride Hepatotoxicity

This study was performed, on the rat model, to form oxidative stress by carbon tetrachloride (CCl4) and to reveal the relationship between toxicity and oxidative/thiol stresses through lipid/protein oxidation and to investigate the effects of N-acetylcysteine on the hepatotoxicity induced oxidative stress. Wistar albino male rats were divided into four groups as CCl4, N-acetylcysteine, CCl4-N-acetylcysteine and Control, each of ten rats. CCl4 (1.0 ml, 800 mg/kg; single dose) and N-acetylcysteine (200 mg; three doses) were intraperitoneally applied to corresponding groups as per kg of rat weight. Hepatotoxicity was identified with histopathological methods. Besides the thiol levels in plasma/tissue samples, pyrrolized protein and total lipid hydroperoxide levels in serum samples were measured. CCl4-induced hepatotoxic lesions remarkably improved in the presence of N-acetylcysteine. There was no significant difference between Control and N-acetylcysteine groups, in terms of parameters measured in serum/plasma and tissue samples. When compared to these groups, pyrrolized protein and total lipid hydroperoxide levels were found to be higher; whereas plasma/tissue thiol levels lower in the CCl4 group. N-acetylcysteine in combination with CCl4 significantly lowered pyrrolized protein and total lipid hydroperoxide levels, and increased thiol levels, so that the values were reached to those of Control and N-acetylcysteine groups. As reflected by higher pyrrolized protein and total lipid hydroperoxide, and lower thiol levels, enhanced free radical production in the CCl4 hepatotoxicity may lead to oxidative and thiol stresses mediated lipid/protein oxidation. In addition, N-acetylcysteine, a powerful antioxidant, may be added as a thiol source to the treatment protocols of several diseases whose pathogenesis is of oxidative and thiol stress.

Гликогенсберегающие свойствасредств растительного происхождения как фактор неспецифической защиты от гепатотоксического поражения ccl4в условиях интенсивной физической нагрузки

The medical-preventive administration of ginseng and eleutherococcus was studied on sub-acute carbon tetrachloride-induced mice liver injury under exhaustive physical activity. The ability of these drugs to reduce CCL4-induced decrease in health expectancy indicator of swimming mice with 7% of the cargo was identified. In this study eleutherococcus and ginseng mostly showed protective effect against a physical endurance than the comparison drug, karsil. To assess the content of glycogen in the liver tissue was carried out histo-chemical study by PAS-reaction. The ranges of reduced glycogen have pale pink color on micrographs, unlike hepatocytes with normal glycogen having deeply stained reddish color that the more intensively, the greater the amount of glycogen contained in the hepatocyte. In all groups of experimental animals treated with CCL4, is uneven loss of glycogen by hepatocytes, which correlates with the data on the mechanism of hepatotoxicity of carbon tetrachloride. There was a significant effect of the phytoadaptogens of the distribution of glycogen under these experimental conditions. This effect may be related to the hepato-protective effect of the drugs, as well as the previously identified ability phyto-adaptogens optimize bioenergetic processes during physical exercises.

β-sitosterol protects against carbon tetrachloride hepatotoxicity but not gentamicin nephrotoxicity in rats via the induction of mitochondrial glutathione redox cycling.

Previous findings have demonstrated that β-sitosterol (BSS), an active component of Cistanches Herba, protected against oxidant injury in H9c2 cardiomyocytes and in rat hearts by enhancing mitochondrial glutathione redox cycling, possibly through the intermediacy of mitochondrial reactive oxygen species production. We therefore hypothesized that BSS pretreatment can also confer tissue protection against oxidant injury in other vital organs such as liver and kidney of rats. In this study, the effects of BSS pretreatment on rat models of carbon tetrachloride (CCl4) hepatotoxicity and gentamicin nephrotoxicity were investigated. The findings showed that BSS pretreatment protected against CCl4-induced hepatotoxicity, but not gentamicin nephrotoxicity in rats. The hepatoprotection afforded by BSS was associated with the improvement in mitochondrial glutathione redox status, presumably through the glutathione reductase-mediated enhancement in mitochondrial glutathione redox cycling. The hepatoprotection afforded by BSS was also accompanied by the improved mitochondrial functional ability in rat livers. The inability of BSS to protect against gentamicin nephrotoxicity was likely due to the relatively low bioavailability of BSS in rat kidneys. BSS may serve as potential mitohormetic agent for the prevention of oxidative stress-induced injury in livers.

An ursolic acid-enriched extract of Cynomorium songaricum protects against carbon tetrachloride hepatotoxicity and gentamicin nephrotoxicity in rats possibly through a mitochondrial pathway: A comparison with ursolic acid

Cynomorium songaricum, which is an herbal tonic for treating impotence in Chinese medicine, serves as a popular health-promoting or invigorating food in China, particularly in the Inner Mongolia Province. Recent studies have shown that HCY2, an active fraction of C. songaricum, and its major ingredient ursolic acid (UA), protect against myocardial ischemia/reperfusion injury in rats. In the present study, we further investigated the effects of HCY2 or UA on carbon tetrachloride (CCl4) hepatotoxicity and gentamicin nephrotoxicity in rats. The results indicated that HCY2 pretreatment protected against CCl4 hepatotoxicity and gentamicin nephrotoxicity, as evidenced by the significant inhibition of plasma aspartate aminotransferase and alanine aminotransferases activities as well as decreases in plasma creatinine and blood urea nitrogen levels, respectively. The hepatoprotection and nephroprotection were associated with the improvement in mitochondrial functional ability, as assessed by the measurement of ATP generation capacity, as well as the enhancement in glutathione redox status, possibly through the induction of mitochondrial uncoupling in rat liver and kidney tissues. However, UA pretreatment at doses equivalent to those of HCY2 was found to protect against CCl4 hepatotoxicity but not gentamicin nephrotoxicity in rats. The discrepancy between HCY2 and UA may be related to the bioavailability of UA to kidney tissue in rats.

Nigella sativa L. oil protects against induced hepatotoxicity and improves serum lipid profile in rats.

The effects of 4 weeks oral intake of Nigella sativa L. (NS) oil on some liver function tests and D-galactosamine- or carbon tetrachloride-induced hepatotoxicity were investigated in male albino rats. In another series of experiments, the effect of the oil on serum lipid profile was examined in male spontaneously hypertensive rats of stroke prone strain and Wistar Kyoto rats. The study showed that daily administration of the oil per se (800 mg/kg orally for 4 weeks) did not adversely effect the serum transaminases (ALT and AST), alkaline phosphatase, serum bilirubin or prothrombin activity in normal albino rats. When the oil was given for 4 weeks prior to induction of hepatotoxicity by D-galactosamine or carbon tetrachloride, it was able to give complete protection against d-galactosamine and partial protection against carbon tetrachloride hepatotoxicity. NS oil showed a favourable effect on the serum lipid pattern where the administration of the oil (800 mg/kg orally for 4 weeks) caused a significant decrease in serum total cholesterol, low density lipoprotein, triglycerides and a significant elevation of serum high density lipoprotein level.

Hepatoprotective effect of &lt;i&gt;euphorbia hirta&lt;/i&gt; on carbon tetrachloride induced liver injury in rats

The aim of the present study was to investigate the hepatoprotective effects of alcoholic extract of Euphorbia hirta on carbon tetrachloride induced liver injury in albino rats. Intraperitoneal injection of carbon tetrachloride (1 ml/kg body weight every 72 hrs for 16 days significantly increased serum pyruvate transaminase, serum oxalate transaminase and serum bilirubin levels in rats. Histopathologically it produced hepatic necrosis, especially in the central lobular region. Simultaneous treatment with Euphorbia hirta (100 and 200 mg/kg body weight daily) and carbon tetrachloride, caused improvement of both biochemical and histopathological parameters compared to that of carbon tetrachloride treatment group alone. However, the extract exhibited significant (p < 0.001) hepatoprotective activity at 200 mg/kg p.o, which was comparable to the activity exhibited by the standard reference silymarin in carbon tetrachloride hepatotoxicity model. These findings were confirmed by the results of the histopathological study of liver.

Hepatoprotective Action of Dehydrozingerone in Carbon Tetrachloride and Thioacetamide-induced Hepatotoxicity in Wistar Rats

© 2011, INASL 40 Synthesis and Antihepatotoxic Activity of 5-(2,3-dihydro-1,4-benzodioxane-6-yl)-3-substitutedphenyl-4,5-dihydro-1H-pyrazole Derivatives Habibullah*,**, B Ahmed** *Department of Pharmaceutical Chemistry, Alwar Pharmacy College, Alwar, Rajasthan **Antihepatotoxic Research Laboratory, Faculty of Pharmacy, Jamia Hamdard, New Delhi Background and Objective: Silymarin isolated from seeds of Silybum marianum commonly known as ‘milk thistle’ has been found most potent antihepatotoxic agent against a variety of toxicants. The Silymarin has been found to be a mixture of three isomers of flavonolignan i.e., silybin, silychristin and silydianin. The silybin is the most potent component containing 1,4-dioxan ring system. Other isomer silychristin and silydianin do not possess 1,4-dioxan ring system and thus, do not display significant antihepatotoxic activity. Based on the above hypothesis we learned that 1,4-dioxane ring system plays an important role in exhibiting antihepatotoxic activity, and hence we have prepared some new pyrazoline derivatives bearing 1,4-dioxane ring system and evaluated them for antihepatotoxic activity against CCl4induced hepatotoxicity in rats. Material and Methods: The titled pyrazolines were prepared by refluxing 2,3-dihydro-1,4-benzodioxane-6-yl)-1-substituted-phenylprop2-en-1-one derivatives with hydrazine hydrate in absolute ethanol and few drops of glacial acetic acid. The synthesized compounds were evaluated for antihepatotoxic activity by measuring different biochemical parameters glutamate oxaloacetate transaminase (GOT) and glutamate pyruvate transaminase (GPT), alkaline phosphatase (ALP) and total protein against CCl4-induced hepatotoxicity in Wistar albino rats as well as histopathological study of the liver. Results: The administration of silymarin (standard drug) and synthesized compounds at a dose level of 10 mg/Kg body weight, prevented CCl4-induced elevation of SGOT, SGPT, ALP, and prevented decrease in total protein. The histopathological studies also showed significant recovery of hepatocytes of the liver in the standard drug and compound-treated animals. Conflict of Interest: None Study of Silymarin on Ethanol Induced Hepatotoxicity and Expression of Bcl-2, Bax and p53 Levels P Vasanth Raj, K Nitesh, P Jain, M Neena Sankhe, J Venkata Rao, C Mallikarjuna Rao, N Udupa Manipal College of Pharmaceutical Sciences, Manipal University, Manipal, Karnataka Background and Aims: Oxidative stress plays a pivotal role in the pathogenesis and progression of alcoholic liver disease (ALD). Considering the antihepatotoxic property of silymarin, a well known antioxidant (flavonoid); we investigated the hepatoprotective effect of silymarin and underlying mechanism(s). Methods: Twenty-four Wistar strain albino rats (180–200 g) of either sex were used. The animals were divided into three groups and treated for 60 days as follows: (1) normal control group received the vehicle (sodium CMC 0.3%); (2) ethanol group was administered ethanol 3.6 g/Kg (30%, 10 mL/Kg) 1 hour after ingestion of sodium CMC 0.3%; (3) ethanol plus silymarin group received silymarin 100 mg/Kg 1 hour prior to the administration of ethanol 3.6 g/Kg. Animals received ethanol and silymarin by oral route daily. Liver damage was evaluated by histopathology and liver function test. At molecular level, expression of P53, Bax and bcl-2 was determined by reverse transcriptase polymerase chain reaction (RT PCR). Results: Silymarin at 100 mg/Kg each ameliorated ethanol-induced macrovesicular steatosis and parenchymatous degeneration in hepatocytes, and decreased serum aminotransferases level. Ethanol downregulated antiapoptotic Bcl-2, upregulated pro apoptotic P53 and Bax mRNA levels in liver hepatocytes, thus, inducing apoptosis. However, this alteration in mRNA levels of P53, Bax and Bcl-2 was significantly prevented by silymarin treated animals at a dose of 100 mg/Kg body weight. Conclusion: Together, these results identify the biological efficacy of silymarin against ethanol-induced hepatotoxicity in rats. Hence, silymarin merits further investigation to support its molecular mechanism. Conflict of Interest: None Hepatoprotective Action of Dehydrozingerone in Carbon Tetrachloride and Thioacetamide-induced Hepatotoxicity in Wistar Rats N Kumar, P Vasanth Raj, A Tiwari, A Chaudhary, C Mallikarjuna Rao, N Gopalan Kutty, N Udupa Manipal College of Pharmaceutical Sciences, Manipal University, Manipal Background and Aims: Dehydrozingerone (DZ) is a prototype constituent of ginger and has shown radioprotection through its antioxidant activity. Chemically DZ is a chalcone in nature and can be prepared easily by reactions of vanillin with acetones. Antioxidant activity 03_JCEH-Abstract.indd 40 3/18/2011 11:13:05 AM

Study of Silymarin on Ethanol Induced Hepatotoxicity and Expression of Bcl-2, Bax and p53 Levels

© 2011, INASL 40 Synthesis and Antihepatotoxic Activity of 5-(2,3-dihydro-1,4-benzodioxane-6-yl)-3-substitutedphenyl-4,5-dihydro-1H-pyrazole Derivatives Habibullah*,**, B Ahmed** *Department of Pharmaceutical Chemistry, Alwar Pharmacy College, Alwar, Rajasthan **Antihepatotoxic Research Laboratory, Faculty of Pharmacy, Jamia Hamdard, New Delhi Background and Objective: Silymarin isolated from seeds of Silybum marianum commonly known as ‘milk thistle’ has been found most potent antihepatotoxic agent against a variety of toxicants. The Silymarin has been found to be a mixture of three isomers of flavonolignan i.e., silybin, silychristin and silydianin. The silybin is the most potent component containing 1,4-dioxan ring system. Other isomer silychristin and silydianin do not possess 1,4-dioxan ring system and thus, do not display significant antihepatotoxic activity. Based on the above hypothesis we learned that 1,4-dioxane ring system plays an important role in exhibiting antihepatotoxic activity, and hence we have prepared some new pyrazoline derivatives bearing 1,4-dioxane ring system and evaluated them for antihepatotoxic activity against CCl4induced hepatotoxicity in rats. Material and Methods: The titled pyrazolines were prepared by refluxing 2,3-dihydro-1,4-benzodioxane-6-yl)-1-substituted-phenylprop2-en-1-one derivatives with hydrazine hydrate in absolute ethanol and few drops of glacial acetic acid. The synthesized compounds were evaluated for antihepatotoxic activity by measuring different biochemical parameters glutamate oxaloacetate transaminase (GOT) and glutamate pyruvate transaminase (GPT), alkaline phosphatase (ALP) and total protein against CCl4-induced hepatotoxicity in Wistar albino rats as well as histopathological study of the liver. Results: The administration of silymarin (standard drug) and synthesized compounds at a dose level of 10 mg/Kg body weight, prevented CCl4-induced elevation of SGOT, SGPT, ALP, and prevented decrease in total protein. The histopathological studies also showed significant recovery of hepatocytes of the liver in the standard drug and compound-treated animals. Conflict of Interest: None Study of Silymarin on Ethanol Induced Hepatotoxicity and Expression of Bcl-2, Bax and p53 Levels P Vasanth Raj, K Nitesh, P Jain, M Neena Sankhe, J Venkata Rao, C Mallikarjuna Rao, N Udupa Manipal College of Pharmaceutical Sciences, Manipal University, Manipal, Karnataka Background and Aims: Oxidative stress plays a pivotal role in the pathogenesis and progression of alcoholic liver disease (ALD). Considering the antihepatotoxic property of silymarin, a well known antioxidant (flavonoid); we investigated the hepatoprotective effect of silymarin and underlying mechanism(s). Methods: Twenty-four Wistar strain albino rats (180–200 g) of either sex were used. The animals were divided into three groups and treated for 60 days as follows: (1) normal control group received the vehicle (sodium CMC 0.3%); (2) ethanol group was administered ethanol 3.6 g/Kg (30%, 10 mL/Kg) 1 hour after ingestion of sodium CMC 0.3%; (3) ethanol plus silymarin group received silymarin 100 mg/Kg 1 hour prior to the administration of ethanol 3.6 g/Kg. Animals received ethanol and silymarin by oral route daily. Liver damage was evaluated by histopathology and liver function test. At molecular level, expression of P53, Bax and bcl-2 was determined by reverse transcriptase polymerase chain reaction (RT PCR). Results: Silymarin at 100 mg/Kg each ameliorated ethanol-induced macrovesicular steatosis and parenchymatous degeneration in hepatocytes, and decreased serum aminotransferases level. Ethanol downregulated antiapoptotic Bcl-2, upregulated pro apoptotic P53 and Bax mRNA levels in liver hepatocytes, thus, inducing apoptosis. However, this alteration in mRNA levels of P53, Bax and Bcl-2 was significantly prevented by silymarin treated animals at a dose of 100 mg/Kg body weight. Conclusion: Together, these results identify the biological efficacy of silymarin against ethanol-induced hepatotoxicity in rats. Hence, silymarin merits further investigation to support its molecular mechanism. Conflict of Interest: None Hepatoprotective Action of Dehydrozingerone in Carbon Tetrachloride and Thioacetamide-induced Hepatotoxicity in Wistar Rats N Kumar, P Vasanth Raj, A Tiwari, A Chaudhary, C Mallikarjuna Rao, N Gopalan Kutty, N Udupa Manipal College of Pharmaceutical Sciences, Manipal University, Manipal Background and Aims: Dehydrozingerone (DZ) is a prototype constituent of ginger and has shown radioprotection through its antioxidant activity. Chemically DZ is a chalcone in nature and can be prepared easily by reactions of vanillin with acetones. Antioxidant activity 03_JCEH-Abstract.indd 40 3/18/2011 11:13:05 AM

Schisandrin B elicits a glutathione antioxidant response and protects against apoptosis via the redox-sensitive ERK/Nrf2 pathway in AML12 hepatocytes

This study examined the effects of (−)schisandrin B [(−)Sch B] on MAPK and Nrf2 activation and the subsequent induction of glutathione antioxidant response and cytoprotection against apoptosis in AML12 hepatocytes. Pharmacological tools, such as cytochrome P-450 (CYP) inhibitor, antioxidant, MAPK inhibitors and Nrf2 RNAi, were used to delineate the signalling pathway. (−)Sch B caused a time-dependent activation of MAPK in AML12 cells, particularly the ERK1/2. The MAPK activation was followed by an enhancement in Nrf2 nuclear translocation and the eliciting of a glutathione antioxidant response. Reactive oxygen species arising from a CYP-catalysed reaction with (−)Sch B seemed to be causally related to the activation of MAPK and Nrf2. ERK inhibition by U0126 or Nrf2 suppression by Nrf2 RNAi transfection almost completely abrogated the cytoprotection against menadione-induced apoptosis in (−)Sch B-pre-treated cells. (−)Sch B pre-treatment potentiated the menadione-induced ERK activation, whereas both p38 and JNK activations were suppressed. Under the condition of ERK inhibition, Sch B treatment did not protect against carbon tetrachloride-hepatotoxicity in an in vivo mouse model. In conclusion, (−)Sch B triggers a redox-sensitive ERK/Nrf2 signalling, which then elicits a cellular glutathione antioxidant response and protects against oxidant-induced apoptosis in AML12 cells.