The Effect Of Compatibility Of Penthorum Chinense Pursh And Flos Puerariae In L-02 Liver Cells Induced By Ethanol

ObjectiveTo explore the effect of compatibility of Penthorum Chinense Pursh and Flos Puerariae containing serum in the L-02 liver cells induced by alcohol and its possible mechanism.MethodsThe clean grade 50 SD rats were divided into 5 groups: compatibility of Penthorum Chinense Pursh and flos puerariae 1:1 group,2:1 group,1:2 group,normal saline group and tiopronin group,were Prepared of containing serum by gavage.L-02 cells Cultured in Vitro were divided into 6 groups: control group,model group,1:1 group,2:1 group,1:2 group,normal saline group and tiopronin group.Matched containing serum were added respectively.After 1 hour all groups were induced by Ethanol 100mmol/L except control group for 24 hours.L-02 cells viability were assessed by MTT assay.Alanine Aminotransferase(ALT)、Aspartate Aminotransferase(AST)、Superoxide Dismutase(SOD)、 Malondialdehyde(MDA)were measured by enzyme label method.The protein and mRNA expression levels of TNF-α、 IL-6、Nrf2 and HO-1 were detected by real-time PCR and Western-blot,so that we can explore the mechanism on liver cells induced by Ethanol and find the best compatibility combination.Results1.Transaminase activity of liver cells induced by Ethanol:Levels of ALT、AST were significantly higher than the control group;each treament group were lower than model group and has a significantly difference compared with model group.However,ALT、AST value among the treament group compared to other group which did not find an outstanding effect.2.Activity of SOD、MDA in liver cells of liver injury :Levels of MDA was significantly higher than the control group,but SOD was lower than it;compared with model group MDA was lower than it in each treament group,SOD was higher and has a significantly difference.However,SOD、MDAvalue among the treament group compared to other group which did not find an outstanding effect.3.In the study of the mechanism,compatibility of Penthorum Chinense Pursh and Flos Puerariae rescription could reduce TNF-α、IL-6 mRNA and protein value,indicating that it could help regulating the balance of inflammatory cytolines.The 1:2 group have a better therapeutic effect on regulating the level of TNF-αprotein than other groups.4.In the study of the mechanism,combination groups could improve Nrf2、HO-1 mRNA and protein value,indicating that it could balance oxidation and anti-oxidation.The 2:1 group had the best effect on improve Nrf2 mRNA value.The 1:1 group had the best effect on improve Nrf2 protein value.Conclusion1.compatibility of Penthorum Chinense Pursh and Flos Puerariae has a therapeutic effect on ethanol-induced hepatocyte injury,The 1:1 group had a litter superiority.2.the therapeutic effect of compatibility of Penthorum Chinense Pursh and Flos Puerariae may affect the expression level of TNF-α,IL-6,Nrf2 and HO-1 to and reduce the inflammatory reaction and oxidative stress in alcohol-induced L-02 liver cells.Then play a role of resistance to alcoholic liver damage,and a new pathway may be exist.