Extract Of Averrhoa Carambola L.（Oxalidaceae） Roots Ameliorates CCl₄-Induced Acute Liver Injury In Mice And Hepatic Fibrosis In Rats

Objective: In the current study,we aim to evaluate the potential therapeutic effect of the extract of Averrhoa carambola L.(Oxalidaceae)roots(EACR)on carbon tetrachloride(CCl4)induced acute liver injury in mice and hepatic fibrosis in rats and to explore the underlying molecular mechanism.Methods: 1.Extract of Averrhoa Carambola L.(Oxalidaceae)Roots Ameliorates CCl4-Induced Acute Liver Injury in Mice 60 Kunming mice were ramdomly separated 6 groups in the study.The vehicle group and model control group received normal saline(NS).The positive group was given Bifendate.And the high,middle,low doses of Extract of Averrhoa Carambola L.Root(EACR)were treated with EACR.Both NS and the treatment protocol were administered via intragastric gavage(i.g.)for 7 days.After 1 hour of the last administration,each group of mice was intraperitineally injected with 0.15% CCl4(10 ml/kg)to establish an acute liver injury model except for the vehicle group.At the end of the experiment,biomarkers assay of liver function and apoptosis,hepatic oxidative stress and antioxidant status and liver tissue histopathological examine were performed to investigate the liver damage.The expression levels of tumor necrosis factor-α(TNF-α),nuclear factor-kappa B(NF-κb),cysteinyl aspartate specific proteinase-3(caspase-3)were measured by western blot.2.Extract of Averrhoa Carambola L.(Oxalidaceae)Roots Ameliorates CCl4-Induced Hepatic Fibrosis in Rats Sprague Dawley rats were randomly divided into six groups of fifteen each as follows: vehicle group,I,II,III,IV,V CCl4 groups.From the first to eighth weeks,CCl4 groups rats were given 50% CCl4 to establish liver fibrosis model.At the sixth,seventh and eighth week,we choose one rat from the vehicle group and CCl4 groups to investigate fibrosis degree respectively.At ninth week,CCl4 groups were randomly divided into five groups fourteen each as follows: CCl4+colchicine group,CCl4+EACR 1.0 g/kg group,CCl4+EACR 0.5 g/kg group and CCl4+EACR 0.25 g/kg group.At the end of the experiment,biomarkers assay of liver function,liver fibrosis,hepatic oxidative stress and antioxidant status,liver tissue histopathological examine and immunostaining were performed to investigate the liver damage and fibrosis degree.The expression levels of type I collagen(Col-1a1),alpha smooth muscle actin(α-SMA),transforming growth factor(TGF-β1),Smad2,Smad4,Smad7 and tissue inhibitor of metalloproteinase 2(TIMP2)in liver tissue were detected by quantitative real time PCR and Western blot.The apoptosis-related proteins(Bax,Bcl2,cleaved Caspase-3 and Caspase-3)in liver tissue were also measured using Western blot.Results: 1.Extract of Averrhoa Carambola L.(Oxalidaceae)Roots Ameliorates CCl4-Induced Acute Liver Injury in Mice After pretreatment EACR,the levels of AST,ALT,IL-β1 and IL-6 in serum as well as MDA in the liver were markedly decreased,whereas the activities of SOD,GSH and GSH-Px were increased.The protein expressions of TNF-α,NF-κb and caspase-3 were significantly down-regulated in the EACR groups.HE staining also showed that the injury of liver was mitigated after administration of EACR.2.Extract of Averrhoa Carambola L.(Oxalidaceae)Roots Ameliorates CCl4-Induced Hepatic Fibrosis in Rats EACR treatment reduced CCl4-induced liver injury,fibrogenic response,and the extent of oxidative stress.EACR treatment also significantly reduced the deposition of collagen and the immunostaining of α-SMA and TGF-β1 in the liver tissue of CCl4-treated rats.In addition,treatment with EACR markedly down-regulated the CCl4-induced m RNA expression of Col-1a1,α-SMA,TIMP2,TGF-β1,Smad2 and Smad4 and suppressed the protein expression of α-SMA,TIMP2,TGF-β1,Smad2,Smad3,Smad4,BAX and cleaved caspase-3.Meanwhile,EACR treatment also significantly elevated the m RNA expression of Smad7 and the protein expression of Smad7 and Bcl-2.Conclusion: EACR can prevent CCl4-induced acute liver injury in mice and against CCl4-induced liver fibrosis.The mechanism may be related to attenuating free radical,inhibiting the lipid peroxidation,apoptosis-inhibition and with modulation of TGF-β1/Smad signaling pathway.