Protective Effect Of Arctigenin On Acute Liver Injury Induced By Concanavalin A In Mice

Objective: The purpose of this study is to explore the effects of arctigenin(ATG)on concanavalin A(ConA)induce acute liver injury of mice at different time points.We investigate the protective effect of ATG on acute liver injury and explore its mechanism.Methods: The C57BL/6 mice were assigned into three groups at randomly: DMSO group,ConA/DMSO group and ConA/ATG group,with 7 mice in each group.DMSO group and ConA/DMSO group were injected with DMSO,ConA/ATG group were given equal volumes of ATG(30mg/kg)by intraperitoneal injection.Five hours later,the mice were injected ConA(20mg/kg)by the tail vein to induce liver injury.Then the mice were killed,blood and the liver tissue were gathered at 0,2,6,12 hours respectively.(1)Detection of serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)activity.(2)HE staining was used to observe liver tissue.(3)Observing the effect of ATG on the survival time of ConA(25mg/kg)induced liver injury at lethal dose.(4)Serum tumor necrosis factor-α(TNF-α),interferon-γ(IFN-γ)and interleukin-12(IL-12)levels were determined by ELISA.(5)Expression of TNF-α,IFN-γ,Bax and Bcl-2 mRNA levels in liver tissue detected by RT-PCR.(6)TUNEL assay was used to detect hepatocyte apoptosis.(7)Flow cytometry was adopted to assess lymphocytic infiltration in liver and spleen.(8)Detection of liver superoxide dismutase(SOD),malondialdehyde(MDA)and myeloperoxidase(MPO).Results:(1)Compared with the ConA/DMSO group,the ATG pretreatment group significantly decreased the levels of serum ALT and AST at 6 and 12 hours(P<0.05).(2)Compared with the ConA/DMSO group,The degree of liver injury was alleviated obviously in ATG pretreatment group.(3)Compared with the ConA/DMSO group,The survival time of mice was obviously prolonged in ATG pretreatment group.(4)Compared with the DMSO group,the levels of serum TNF-α,IFN-γ and IL-12 were increased significantly in ConA/DMSO group;Compared with the ConA/DMSO group,the ATG pretreatment group decreased the levels of serum TNF-α,IFN-γ at 6h and 12 h,and decreased IL-12 level at 2h,6h and 12h(P<0.05).(5)Compared with the DMSOgroup,the expression of TNF-α、IFN-γ and proapoptotic gene Bax mRNA in liver tissue were increased and antiapoptotic gene Bcl-2 mRNA was reduced in ConA/DMSO group;Compared with the ConA/DMSO group,the expression of TNF-α 、 IFN-γ and Bax m RNA were reduced and Bcl-2 mRNA was increased(P<0.05).(6)Compared with the DMSO group,the number of hepatocytes apoptosis was increased in the ConA/DMSO group;Compared with the ConA/DMSO group,the ATG pretreatment group decreased the number of hepatocytes apoptosis.(7)Compared with the ConA/DMSO group,the ATG pretreatment group enhanced recruitment of MDSCs,and affected the ratio of T cells,NK cells and NKT cells in liver and spleen.(8)Compared with the ConA/DMSO group,the ATG pretreatment group increased SOD activity at 24h(P<0.05),decreased MDA activity at 2h,12 h and 24h(P<0.05),and enhanced MPO activity at 6h and 12h(P<0.01).Conclusion: The model of acute liver injury induced by ConA was successfully constructed in mice.It was proved that ATG had obvious protective effect on acute liver injury induced by ConA.The protective effect of ATG was mainly by inhibiting the expression and release of inflammatory factors,blocking the apoptotic pathway of hepatocyte,promoting recruitment of MDSCs,and inhibiting lipid peroxidation of the hepatocyte membrane.