Ameliorative effects of phyllanthin on carbon tetrachloride-induced hepatic oxidative damage in mice

ObjectiveTo evaluate the liver protecting efficacy of phyllanthin, a lignin, isolated from the leaves of Phyllanthus amarus using mice model.MethodsPhyllanthin was orally administered with or without CCl₄ for 30 d. Serum levels of hepatic marker enzymes namely alanine transaminase and aspartate transaminase were evaluated. Oxidative stress was ascertained by measuring hepatic lipid peroxidation levels and by estimating non-enzymatic antioxidants such as glutathione, total ascorbic acid, enzymatic antioxidants namely catalase, superoxide dismutase, glutathione peroxidase, glutathione reductase, and glutathione transferase. Histopathological and ultramicroscopic analyses were also carried out.ResultsOral administration of CCl₄ caused significant increase in lipid peroxidation. The hepatic levels of both non-enzymatic antioxidants and enzymatic antioxidants were significantly lowered in CCl₄-treated mice as compared to control. Treatment with phyllanthin significantly mitigated these changes in the CCl₄-treated mice. Histopathological and ultramicroscopic studies correlated well with the biochemical findings, as phyllanthin treatment reversed the alterations induced by the toxin and the subcellular features of phyllanthin treated mice were similar to those present in the normal mouse liver.ConclusionsThis study reports the in vivo anti-hepatotoxic potential of this isolated molecule phyllanthin, which may be responsible for the liver protecting property of Phyllanthus amarus.     

Evaluation of hepatoprotective activity of alcoholic and aqueous extracts of Selaginella lepidophylla

ObjectiveTo screen Selaginella lepidophylla (S. lepidophylla) that are used in traditional medicine for their claimed hepatoprotective properties.MethodsAlcoholic and aqueous extracts of S. lepidophylla were evaluated for their hepatoprotective activity using CCl₄ and paracetamol induced acute hepatic injury model.ResultsTreatment with CCl₄ and paracetamol significantly increased liver weight and volume compared to the normal group. Pretreatment with silymarin alcoholic and aqueous extracts significantly prevent increase in liver weight and volume.ConclusionsFrom the present experimental study it can be concluded that alcoholic and aqueous extract of S. lepidophylla exhibited significant hepatoprotective activity against CCl₄ and paracetamol induced hepatotoxicity in rats, as result showed in physical, biochemical and histopathological parameters.     

Efficacy evaluation of the protein isolated from Peganum harmala seeds as an antioxidant in liver of rats

ObjectiveTo evaluate the curative effect of the 132 KD protein isolated from the seeds of Peganum harmala (P. harmala) L. against carbon tetrachloride (CCl₄) induced oxidative stress in rats.MethodsAnimals were post treated intraperitoneally with 132 KD isolated protein at doses of 4 and 8 mg/kg body weight and bovine serum albumin (BSA) (8 mg/kg body weight) as well as vitamin C (250 mg/kg body weight p.o.) for 7 d after they challenged with CCl₄ orally (1 mL/kg body weight) in olive oil (50%) for 2 d.ResultsThe purified protein from seeds of P. harmala plant showed in vitro antioxidant activity with DPPH assay. Administration of CCl₄ induced induction in serum aminotransferases (AST, ALT), alkaline phosphatase (ALP), lipid profile parameters and liver malondialdehyde (MDA) and decrease in serum total protein, liver superoxide dismutase (SOD), catalase (CAT), and reduced glutathione (GSH) levels. 132 KD protein treatment of rats post CCl₄ intoxication successfully alleviated the toxic effects of CCl₄.ConclusionsThe isolated protein possessed strong antioxidant activity comparable to that of BSA (negative control) and vitamin C (positive control).     

Antioxidant,hepatoprotective and hypolipidemic effects of methanolic root extract of Cassia singueana in rats following acute and chronic carbon tetrachloride intoxication

ObjectiveTo evaluate in vivo antioxidant and hepatoprotective activities of the methanolic extract of the root of Cassia singueana in rats following acute and chronic carbon tetrachloride intoxication.MethodsMalondialdehyde (MDA), aspartate aminotransferase (AST), alanine aminotransferase (ALT) and bilirubin as indices of liver damage and lipid peroxidation were detected in rats after intraperitoneal administration of extract (5 mg/kg).ResultsThe liver, kidney and heart showed significant reduction (P<0.05) in the levels of MDA from (0.18±0.04), (0.23±0.07) and (0.26±0.10) nmol/mg respectively in the CCl₄ control to (0.15±0.03), (0.17±0.04) and (0.17±0.07) nmol/mg protein in groups pre-treated with the extract for three days at 5 mg/kg). Similarly, compared to the CCl₄ control, significant reduction (P<0.05) in serum AST, ALT and bilirubin as well as in level of total cholesterol and MDA with concomitant increase in HDL cholesterol, superoxide dismutase and catalase levels when CCl₄-intoxicated rats were treated with Cassia singueana root extract for two weeks.ConclusionsThese results suggest that methanolic extract of Cassia singueana contain potent antioxidant compounds that can offer significant protection against hepatic and oxidative injuries.     

An aqueous extract of Stevia rebaudiana variety Morita II prevents liver damage in a rat model of cirrhosis that mimics the human disease

Introduction and aimStevia has exhibited antioxidant, antihyperglycemic, antihypertensive and anti-inflammatory properties in several in vivo and in vitro models. The objective of this study was to investigate the ability of an aqueous extract of stevia (AES) to prevent experimental cirrhosis in rats and to explore its mechanism of action.Materials and methodsLiver cirrhosis was induced by administering carbon tetrachloride (CCl₄) (400 mg/kg by i.p. injection 3 times a week for 12 weeks); AES was administered (100 mg/kg by gavage daily) during the CCl₄ treatment. Fibrosis was evaluated with histological, biochemical and molecular approaches, and liver damage was assessed with standardized procedures. The profibrotic pathways were analyzed by western blotting, qRT-PCR and immunohistochemistry.Results and conclusionsChronic CCl₄ administration increased nuclear factor kappa B (NF-κB) and proinflammatory cytokine production as well as oxidative parameters such as lipid peroxidation and 4-hydroxynonenal levels, whereas GSH and nuclear factor-E2-related factor 2 (Nrf2) levels were decreased. CCl₄ induced profibrogenic mediator expression, hepatic stellate cell (HSC) activation and, consequently, extracellular matrix production. AES exhibited antioxidant, anti-inflammatory and antifibrotic properties, probably because of its capacity to induce Nrf2 expression, reduce NF-κB expression and block several profibrogenic signaling pathways, subsequently inhibiting HSC activation and preventing fibrosis induced by chronic CCl₄ administration.     

Restorative effect of Eclipta alba in CCl4 induced hepatotoxicity in male albino rats

ObjectiveTo elucidate the restorative effect of the aqueous leaf extract (85%) of the traditional medicinal plant Eclipta alba (E. alba) against CCl₄ induced hepatotoxicity in male albino rats.MethodsRestorative activity was assessed using CCl₄-induced hepatic injury in rats by monitoring biochemical parameters. Biochemical markers of hepatic damage such as aspartate transaminase (AST), alanine transaminase (ALT) and alkaline phosphatase (ALP) were assessed in serum. The oxidative stress was evaluated by measuring the levels of thiobarbutric acid reactive substance (TBARS), hydroperoxides, activity levels of enzymes viz. superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPX) and glutathione-s-transferase (GST) in hepatic tissue.ResultsCCl₄ and olive oil mixture (1:1 dosage of 1 mL/kg b.w.) induced oxidative stress was indicated by elevated levels of TBARS and hydroperoxides, and augmented levels of serum AST, ALT and ALP. The depleted activity levels of antioxidant enzymes such as SOD, CAT, GPX and GST were found in CCl₄ induced animals. The aqueous leaf extract of E. alba (250 mg/kg b.w.) ameliorated the effects of CCl₄ and returned the alter levels of the biochemical markers near to the normal levels.ConclusionsThe study indicates that aqueous leaf extract of E. alba has potential restorative effect on CCl₄ induced hepatotoxicity in male albino rats.     

Hepatoprotective and antioxidant capacity of Melochia corchorifolia extracts

ObjectiveTo evaluate hepato protective and antioxidant capacity of Melochia corchorifolia (M. corchorifolia) aerial part extracts.MethodsAntioxidant activity was evaluated by using three free radicals (Superoxide, Hydroxyl and DPPH) and hepatoprotective activity was assessed against CCl₄ induced liver intoxication in rats.ResultsThe extracts produced concentration dependent percentage protection in decrease of serum enzymes and percentage inhibition on free radicals. Among all extracts methanol extract showed better activity with percentage protection of SGOT (78.98%), SGPT (79.65%), ALP (82.48%) and total bilirubin (80.0%) levels against CCl₄ liver intoxication and also methanolic extract showed better activity with IC₅₀ values on superoxide, hydroxyl and DPPH radicals were 127 μ g, 240 μ g and 179 μ g.ConclusionsFrom the results obtained during the study it could be concluded that M. corchorifolia aerial part extracts have antioxidant and hepatoprotective components. Further study is necessary for isolation and characterization of bioactive molecules which are responsible for hepatoprotective and antioxidant activity.     

Antipyretic, anti-inflammatory and analgesic properties of nilavembu kudineer choornam: a classical preparation used in the treatment of chikungunya fever

ObjectiveTo investigate the efficacy of ethanolic extract of nilavembu kudineer choornam (EENKC) in inflammation, pain and fever using animal models to support its actions.MethodsAcute toxicity study of EENKC was performed in mice to fix the effective dose. The antipyretic, anti-inflammatory and analgesic activity of EENKC was evaluated in brewer's yeast induced pyrexia in rats, carrageenan-induced inflammation in rats and acetic-acid induced writhing in mice model.ResultsAcute toxicity revealed that EENKC didn't show death and toxic signs up to 2 000 mg/kg. In brewer's yeast induced pyrexia and carrageenan-induced inflammation EENKC at the doses of 200 and 400 mg/kg inhibited fever and inflammation significantly (P<0.01 and <0.05) compared to control animals. In mice, the number of writhing induced by acetic-acid was significantly (P<0.01) reduced after treatment with both the dose of EENKC than control animals. EENKC 200 mg/kg inhibits inflammation higher level in carrageenan-induced paw edema, but there is no significant difference when compared to indomethacin 10 mg/kg.ConclusionsThe present findings revealed that EENKC possesses antipyretic, anti-inflammatory and analgesic activity which supports nilavembu kudineer choornam efficacy in chikungunya fever.     

Pharmacokinetics of acemetacin and its active metabolite indomethacin in rats during acute hepatic damage and liver regeneration

Background and aim:The pharmacokinetics of acemetacin, a non-steroidal anti-inflammatory drug which is biotransformed to indomethacin by hepatic first-pass effect, was examined during the necrotic and regeneration phases resulting from acute hepatitis induced by carbon tetrachloride (CCl₄).Material and methods: Acute hepatitis was induced by oral CCl₄ administration to male Wistar rats. On days 0, 1 and 3 after the insult, liver histological analysis was performed, biochemical markers of liver damage and regeneration were measured, and the pharmacokinetics of oral acemetacin and of its active metabolite, indomethacin, were determined.Results: One day after CCl₄ administration, liver necrosis was apparent and there was an increase in the circulating levels of indicators of liver damage and regeneration with regard to control conditions. Acemetacin bioavailability was increased, although not in a statistically significant manner. On the other hand, indomethacin bioavailability was significantly reduced. By day 3, histological analysis revealed liver recovery, although not complete, while biochemical indicators of hepatic damage had reverted either totally or partially. Markers of liver regeneration were still increased. Bioavailability acemetacin and indomethacin was comparable to control values.In conclusion: Indomethacin bioavailability after oral administration of its precursor, acemetacin, is significantly reduced by acute hepatitis produced by CCl₄. Pharmacokinetic alterations, as liver damage, are reversible, but do not require complete liver regeneration to return to basal conditions.     

Analgesic and anti-inflammatory activities of Passiflora foetida L

ObjectiveTo investigate the analgesic and anti-inflammatory activities of ethanol extract of Passiflora foetida (P. foetida) leaves.MethodsEthanol extract of P. foetida leaf was evaluated for analgesic action by acetic acid-induced writhing and hot plate method in albino mice. The anti-inflammatory property of ethanolic leaf extract was tested by carrageenan induced acute paw edema and histamine induced acute paw edema in rats.ResultsThe dose 200 mg/kg of P. foetida leaf extract exhibited highest significant analgesic activity [(13.50±0.43) min] at a reaction time of 20 min in hot plate method in mice. The ethanol extract of leaf dose 100 mg/kg produced a highly significant anti inflammatory effect [(1.302±0.079) mL] in rats.ConclusionsIt is very clear that P. foetida also has analgesic and anti-inflammatory activities for the pharmaceuticals.     

Protective effect of Sida cordata leaf extract against CCl4 induced acute liver toxicity in rats

ObjectiveTo investigate the hepatoprotective potential of Sida cordata (Malvaceae) (S. cordata) in experimental rats to validate its traditional claim.MethodsWister albino rats were divided into 6 groups: Group I served as control; Group II served as hepatotoxic (CCl₄ treated) group; Group III, IV and V served as (100, 200 and 400 mg/kg b.w.) S. cordata leaf extract (SCLE) treated groups; Group VI served as positive control (Silymarin) treated group. Liver marker enzymes serum glutamate oxyloacetic transaminase, serum glutamic pyruvic transaminase, pancreatic enzymatic antioxidants superoxide dismutase (SOD), lipid peroxidation, catalase (CAT), reduced glutathione (GSH) were measured and compared along with histopathological studies.ResultsObtained results show that the treatment with SCLE significantly (P<0.05-<0.001) and dose-dependently reduced CCl₄ induced elevated serum level of hepatic enzymes. Furthermore, SCLE significantly (up to P<0.001) reduced the lipid peroxidation in the liver tissue and restored activities of defence antioxidant enzymes GSH, SOD and CAT towards normal levels, which was confirmed by the histopathological studies.ConclusionsThe results of this study strongly indicate the protective effect of SCLE against CCl₄ induced acute liver toxicity in rats and thereby scientifically support its traditional use.     

Hepatoprotective effect of Premna corymbosa (Burm. f.) Rottl. & Willd. leaves extract on CCl4 induced hepatic damage in Wistar albino rats

ObjectiveTo investigate the hepetoprotective activity of Premna corymbosa leaves against carbon tetrachloride (CCl₄) induced hepatic damage.MethodsHepatotoxicity was induced in wistar rats of both sexes by intraperitoneal injection of CCl₄, 1 mL/kg body weight for every 72 h. The ethanolic extract of Premna corymbosa leaves were administrated at doses of 200 & 400 mL/kg body weight, p. o., daily for 14 days. The hepatotoxicity and its prevention was assessed by serum markers like serum alkaline phosphatase (SALP), serum triglycerides (STG), serum total protein (STP), serum cholesterol (SC), and liver wet weight and histopathological studies of the liver.ResultsIn treatment with the ethanolic extract, the toxic effect of CCl₄ was controlled significantly (P < 0.01) by restoration of the levels of biochemical parameters as compared to normal and standard drug silymarin treated groups. The liver weight was reduced by the ethanolic extract treated groups. The histopathology of the liver sections evidenced the hepatoprotective activity.ConclusionThe ethanolic extract of the leaves of Premna corymbosa possess significant acute hepatoprotective activity. Premna corymbosa can be recommended for the liver disorders.     

Hepatoprotective effect of the selective mineralocorticoid receptor antagonist, eplerenone against carbon tetrachloride-induced liver injury in rats

Background. Eplerenone is a selective mineralocorticoid receptor (MR) antagonist, and its potential protective role in cardiovascular injury has been reported by several studies. However, whether and how this drug can ameliorate hepatic injury in rats is unknown.Material and methods. The present study was conducted to investigate effect of eplerenone against liver injury induced by carbon tetrachloride (CCl₄) in rats. The biochemical liver function tests and oxidative stress parameters including malondialdehyde (MDA), reactive oxygen species (ROS), in addition to the reduced glutathione (GSH) levels were evaluated. Moreover, serum tumor necrotic factors (TNF-α) level and histopathological changes were examined.Results. Our results show that pre-treatment with eplerenone (4 mg/kg per day for 4 weeks) revealed attenuation in serum activities of alanine aminotransferase (ALT), aspartate aminotransferase, (AST), alkaline phosphatase (ALP) and bilirubin levels that were enhanced by CCl₄. Further, pre-treatment with eplerenone inhibited the elevated hepatic MDA content and restored hepatic GSH to its normal level. The enhanced hepatic ROS production in CCl₄-treated group was markedly decreased by eplerenone administration. Eple-renone pre-treatment significantly attenuated the inflammatory responses caused by CCl₄ as evident by the decreased serum TNF-α level. Histopathological studies showed that eplerenone alleviated the liver damage and reduced the lesions caused by CCl₄.Conclusion. Collectively, the present study provides a proof to hepatoprotective actions of eplerenone via reducing oxidative stress and inflammatory responses in CCl₄-induced liver damage in rat model.     

Hepatoprotective and antioxidant activity of methanolic extract of flowers of Nerium oleander against CCl4-induced liver injury in rats

ObjectiveTo investigate the antioxidant and hepatoprotective activity of methanolic flower extract of Nerium oleander against CCl₄-induced hepatotoxicity in rats.MethodsIn vitro antioxidant activity of methanolic extract of flowers of Nerium oleander (MENO–F) was evaluated by various assays, including reducing power, lipid peroxidation, DPPH, ABTS, superoxide anion, hydroxyl radicals and metal chelation. The hepatoprotective and in vivo antioxidant activity of MENO-F were evaluated against CCl₄–induced hepatic damage in rats. The MENO-F at dose of 100, 200 and 400 mg/kg were administered orally once daily for seven days. Serum enzymatic levels of serum glutamate oxaloacetate transaminase (AST), serum glutamate pyruvate transaminase (ALT), serum alkaline phosphatase (ALP) and total bilirubin were estimated along with estimation of superoxide dismutase (SOD) and malondialdehyde (MDA) levels in liver tissues. Further histopathological examination of the liver sections was carried out to support the induction of hepatotoxicity and hepatoprotective efficacy.ResultsThe extract showed potent activities on reducing power, lipid peroxide, DPPH, ABTS, superoxide anion, hydroxyl radical and metal chelation. The substantially elevated serum enzymatic levels of AST, ALT, ALP and total bilirubin were found to be restored towards normalization significantly by the MENO-F in a dose dependent manner with maximum hepatoprotection at 400 mg/kg dose level. The histopathological observations supported the biochemical evidences of hepatoprotection. Elevated level of SOD and decreased level of MDA further strengthen the hepatoprotective observations.ConclusionsThe results of the present study strongly reveal that MENO-F has potent antioxidant activity and hepatoprotective activity against CCl₄-induced hepatic damage in experimental animals.     

Extracellular histones stimulate collagen expression in vitro and promote liver fibrogenesis in a mouse model via the TLR4-MyD88 signaling pathway

BACKGROUNDLiver fibrosis progressing to liver cirrhosis and hepatic carcinoma is very common and causes more than one million deaths annually. Fibrosis develops from recurrent liver injury but the molecular mechanisms are not fully understood. Recently, the TLR4-MyD88 signaling pathway has been reported to contribute to fibrosis. Extracellular histones are ligands of TLR4 but their roles in liver fibrosis have not been investigated.AIMTo investigate the roles and potential mechanisms of extracellular histones in liver fibrosis.METHODS In vitro, LX2 human hepatic stellate cells (HSCs) were treated with histones in the presence or absence of non-anticoagulant heparin (NAHP) for neutralizing histones or TLR4-blocking antibody. The resultant cellular expression of collagen I was detected using western blotting and immunofluorescent staining. In vivo, the CCl₄-induced liver fibrosis model was generated in male 6-week-old ICR mice and in TLR4 or MyD88 knockout and parental mice. Circulating histones were detected and the effect of NAHP was evaluated.RESULTSExtracellular histones strongly stimulated LX2 cells to produce collagen I. Histone-enhanced collagen expression was significantly reduced by NAHP and TLR4-blocking antibody. In CCl₄-treated wild type mice, circulating histones were dramatically increased and maintained high levels during the duration of fibrosis-induction. Injection of NAHP not only reduced alanine aminotransferase and liver injury scores, but also significantly reduced fibrogenesis. Since the TLR4-blocking antibody reduced histone-enhanced collagen I production in HSC, the CCl₄ model with TLR4 and MyD88 knockout mice was used to demonstrate the roles of the TLR4-MyD88 signaling pathway in CCl₄-induced liver fibrosis. The levels of liver fibrosis were indeed significantly reduced in knockout mice compared to wild type parental mice.CONCLUSIONExtracellular histones potentially enhance fibrogenesis via the TLR4–MyD88 signaling pathway and NAHP has therapeutic potential by detoxifying extracellular histones.     

Dihydromyricetin alleviates carbon tetrachloride-induced acute liver injury via JNK-dependent mechanism in mice

AIM: To assess the effects of dihydromyricetin(DHM) as a hepatoprotective candidate in reducing hepatic injury and accelerating hepatocyte proliferation after carbon tetrachloride(CCl4) treatment.METHODS: C57 BL/6 mice were used in this study. Mice were orally administered with DHM(150 mg/kg) for 4 d after CCl4 treatment. Serum and liver tissue samples were collected on days 1, 2, 3, 5 and 7 after CCl4 treatment. The anti-inflammatory effect of DHM was assessed directly by hepatic histology detection and indirectly by serum levels of aspartate aminotransferase(AST), alanine aminotransferase(ALT), albumin, and superoxide dismutase(SOD). Inflammatory cytokines, such as interleukin(IL)-1β, IL-6 and tumor necrosis factor-α(TNF-α), were detected using ELISA kits. Proliferating cell nuclear antigen(PCNA) staining was used to evaluate the role of DHM in promoting hepatocyte proliferation. Hepatocyte apoptosis wasmeasured by TUNEL assay.Furthermore,apoptosis proteins Caspases-3,6,8,and 9 were detected by Western blot.SP600125 were used to confirm whether DHM regulated liver regeneration through JNK/TNF-αpathways.RESULTS:DHM showed a strong anti-inflammatory effect on CCl4-induced liver injury in mice.DHM could significantly decrease serum ALT,AST,IL-1β,IL-6 and TNF-αand increase serum albumin,SOD and liver SOD compared to the control group after CCl4 treatment(P0.05).PCNA results indicated that DHM could significantly increase the number of PCNA positive cells compared to the control(348.9±56.0 vs 107.1±31.4,P0.01).TUNEL assay showed that DHM dramatically reduced the number of apoptotic cells after CCl4 treatment compared to the control(365.4±99.4 vs 90.5±13.8,P0.01).Caspase activity detection showed that DHM could reduce the activities of Caspases-8,3,6 and 9 compared to the control(P0.05).The results of Western blot showed that DHM increased the expression of JNK and decreased TNF-αexpression.However,DHM could not affect TNF-αexpression after SP600125 treatment.Furthermore,DHM could significantly improve the survival rate of acute liver failure(ALF)mice(73.3%vs 20.0%,P0.0001),and SP600125 could inhibit the effect of DHM.CONCLUSION:These findings demonstrate that DHM alleviates CCl4-induced liver injury,suggesting that DHM is a promising candidate for reversing liver injury and ALF.     

Hepatoprotective and antioxidant activity of pentagamavunon-0 against carbon tetrachloride-induced hepatic injury in rats

ObjectiveTo investigate the hepatoprotective and antioxidant activity of pentagamavunon-0(PGV-0) against CCl₄-induced hepatic injury in rats.MethodsThe groups of animals were administered with PGV-0 at the doses 2.5, 5, 10, and 20 mg/kgb.w., p.o. once in a day for 6 days and at day 7 the animals were administrated with carbon tetrachloride (CCl) (20%, 2 mL/kgb.w. in liquid paraffin (i.p.). The effect of PGV-0 on serum transaminase (SGPT), alkaline phosphates (ALP) and total bilirubin were determined in CCl₄-induced hepatotoxicity in rats. Further, the effects of PGV-0 on glutathione (GSH) content, catalase (CAT) and NO free radical scavenging activity also were investigated.ResultsThe results demonstrated that PGV-0 significantly reduced the activity of SGPT, serum ALP and total bilirubin in CCl₄induced rat hepatotoxicity. PGV-0 has effect on the antioxidant and free radical defense system. It prevented the depletion level of GSH and decrease activity of CAT in CCl₄-induced liver injury in rats. PGV-0 also demonstrated the free radical scavenger effects on NO free radical scavenging activity with ES value of 32.32μM.ConculsionAll of our findings suggests that PGV-0 could protect the liver cells from CCl₄-induced liver damages and the mechanism may through the antioxidative effect of PGV-0 to prevent the accumulation of free radicals and protect the liver damage.     

Therapeutic Effects and Possible Mechanisms of a Snake Venom Preparation in the Fibrotic Rat Liver

The effects of a Chinese snake venom preparation from Agkistrodon halys pallas, used for treatment of hepatic fibrosis/cirrhosis in China, was investigated in an {in vivo} rat model and using in situ hepatic perfusion. Four groups were used in the experiments: (i) healthy, (ii) healthy/venom-treated, (iii) carbon tetrachloride (CCl₄)-treated, and (iv) CCl₄/venom-treated. Treatment effects were assessed by determining hepatic histopathology, biochemistry and fibrosis index parameters, bile production, biliary taurocholate recovery, hepatic mRNA expression of four bile salt transporters (Ntcp, Bsep, Oatp-1, and Oatp-3), comparison of hepatic microcirculation, fibrinolytic activity, and antithrombotic effects. Liver histopathology, biochemistry, and fibrosis index showed a dramatic improvement in venom-treated animals. There were significant differences in bile production between healthy/venom-treated and all other experimental groups and between CCl₄/venom-treated and CCl₄-treated animals, but no significant differences were found between CCl₄/venom-treated and healthy animals. Biliary taurocholate recovery was significantly increased in healthy/venom-treated and CCl₄/venom-treated animals. The expression of mRNA levels of the four bile salt transporters showed an increase after venom treatment. The hepatic microcirculation studies showed normalized sinusoidal beds in CCl₄/venom-treated animals compared to healthy animals, whereas CCl₄-treated animals showed abnormal profiles to the healthy and the CCl₄/AHPV-treated animals. The fibrinogen and plasma thromboxane B₂ levels of healthy rats decreased with increasing dose after venom treatment. It was concluded that snake venom treatment may be therapeutic in treatment of hepatic fibrosis/cirrhosis by possibly a combination of increased bile flow and improved hepatic microcirculation, changes in bile salt transporter expression, and fibrinolytic and antithrombotic effects of the snake venom preparation.     

Phycocyanobilin accelerates liver regeneration and reduces mortality rate in carbon tetrachloride-induced liver injury mice

AIM: To investigate the hepatoprotective effects of phycocyanobilin(PCB) in reducing hepatic injury and accelerating hepatocyte proliferation following carbon tetrachloride(CCl4) treatment.METHODS: C57BL/6 mice were orally administered PCB 100 mg/kg for 4 d after CCl4 injection, and then the serum and liver tissue of the mice were collected at days 1, 2, 3, 5 and 7 after CCl4 treatment. A series of evaluations were performed to identify the curative effects on liver injury and recovery. Aspartate aminotransferase(AST), alanine aminotransferase(ALT), albumin and superoxide dismutase(SOD) were detected to indirectly assess the anti-inflammatory effects of PCB. Meanwhile, we detected the expressions of hepatocyte growth factor, transforming growth factor alpha(TGF-α), TGF-β, tumor necrosis factor-alpha(TNF-α) and interleukin-6(IL-6), the factors which are associated with inflammation and liver regeneration. The protein expressions of proliferating cell nuclear antigen(PCNA), TNF-α and cytochrome C were detected by western blot. Furthermore, the survivalrates were analyzed of mice which were administered a lethal dose of CCl4(2.6 mg/kg)with or without PCB.RESULTS:In our research,PCB showed a strongly anti-inflammatory effect on CCl4-induced liver injury in mice.The ALT was significantly decreased after CCl4 treatment from day 1(P0.01)and the AST was significantly decreased from day 2(P0.001).Both albumin and liver SOD were increased from day2(P0.001 and P0.01),but serum SOD levels did not show a significant increase(P0.05).PCB protected the structure of liver from the injury by CCl4.TUNEL assay showed that PCB dramatically reduced the number of apoptotic cells after CCl4 treatment compared to the control(101.0±25.4 vs 25.7±6.4,P0.01).The result of western blotting showed that PCB could increase PCNA expression,decrease TNF-αand cytochrome C expression.Furthermore,data shows that PCB could improve the survival rate of acute liver failure(ALF)mice which were injected with a lethal dose of CCl4(60.0%vs 20.0%).CONCLUSION:Our study indicated that PCB could be an ideal candidate for reversing acute liver injury or ALF.     

Monocyte-derived fibrocytes elimination had little contribution on liver fibrosis

BackgroundMonocyte-derived fibrocytes play an important role in the progression of fibrosis in the skin, lungs, heart and kidney. However, the contribution of fibrocytes to liver fibrosis is unclear. The aim of this study was to investigate whether fibrocytes contributed to fibrosis progression in the livers of carbon tetrachloride (CCl₄)-treated mice.MethodsC57BL/6J mice were divided into 4 groups: normal control group, CCl₄-treated group, CCl₄ + control liposome-treated group, and CCl₄ + clodronate liposome-treated group. For the elimination of systemic monocyte and monocyte-derived fibrocyte, one group was treated with clodronate liposome, and another group with control liposome as a control. After 4 weeks of treatment, hepatic mononuclear cells were subjected to immunofluorescent (IF) staining and fluorescence-activated cell sorter (FACS) analysis to detect fibrocytes. Measurement of collagen-positive Sirius red stained area and collagen-I mRNA expression in the liver were performed to evaluate the degree of liver fibrosis quantitatively.ResultsIn the liver of the CCl₄-treated and CCl₄ + control liposome-treated groups, the number of fibrocytes, the area positive for Sirius red staining and collagen-I mRNA expression significantly increased compared with those in the normal control group. In the liver of the CCl₄ + clodronate liposome-treated group, few fibrocytes was observed as in the normal control group, but Sirius red staining positive area and collagen-I mRNA expression were increased and equivalent to the CCl₄-treated and CCl₄ + control liposome-treated groups.ConclusionMonocyte-derived fibrocytes play a minimal role in CCl₄-induced liver fibrosis. Cells other than fibrocytes such as hepatic stellate cells play a central role in liver fibrosis.