Resveratrol Improves Non-alcoholic Fatty Liver Disease By Promoting The Expression Of The Antioxidant Factor SP1/ZNF32

Part ?: Resveratrol improves non-alcoholic fatty liver disease in model animals by promoting the expression of the antioxidant factor SP1/ZNF32Objective: The incidence of non-alcoholic fatty liver disease(NAFLD)has been significantly increasing in china recently,especially among the youth.At present,it still has no effective approach for its prevention and treatment.In this study,we mainly investigated the preventive and therapeutic effects of resveratrol(RSV)on non-alcoholic fatty liver disease(NAFLD)and the mechanism of action of antioxidant factors SP1 and ZNF32 in the treatment of NAFLD.Methods: A guinea pig model of NAFLD was established by feeding with modified high-fat diet.Guinea pigs were divided into 5 groups: control group,NAFLD model group,high-dose RSV group,low-dose RSV group and simvastatin group.HE staining and oil red O staining were used to assess the changes of fatty content and lipid deposition in the guinea pig liver.The plasma levels of TC,TG,HDL-C,LDL-C,ALT and AST in Guinea pigs were detected by automatic biochemical analyzer.Furthermore,the m RNA expression of SP1,ZNF32,Caspase2 and Caspase9 in guinea pigs were analyzed by RT-PCR.The levels of SOD and MDA in liver tissues of guinea pigs were detected to observe the effect of RSV on oxidative damage in NAFLD guinea pigs.Results: The NAFLD guinea pig model was successfully established by feeding a high-fat diet.The histopathological changes in guinea pigs liver were evaluated through HE and oil red O staining,it was found that the degree of hepatic steatosis in the RSV and simvastatin groups were significantly improved compared with the NAFLD guinea pig model group.The liver function indexes and blood lipids level in the RSV and simvastatin groups were also significantly lower compared with the NAFLD guinea pig model group.The m RNA expression of antioxidant genes SP1 and ZNF32 increased significantly in the RSV and simvastatin groups.The expression of apoptosis genes Caspase2 and Caspase9 in the RSV group increased,but the expression in the simvastatin group decreased.RSV and simvastatin can significantly reduce the content of MDA in liver tissue of NAFLD guinea pigs.At the same time,RSV can significantly increase the activity of SOD in liver tissue of NAFLD guinea pigs.Simvastatin had no significant effect on SOD activity in NAFLD guinea pigs.Conclusions: This study demonstrated that the protective effect of resveratrol on the NAFLD guinea pig model were very significant,and resveratrol exerts an anti-oxidative effect by promoting SP1/ZNF32 expression to retard the NAFLD progression.In summary,resveratrol has a good effect on the prevention and treatment of NAFLD and is expected to be a feasible therapeutic drug for NAFLD.This study provides a theoretical basis for the subsequent clinical studies.Part ?: Establishment of L-02 hepatic steatosis cell model and the preliminary study on protective effect of resveratrol in itObjective: To induce steatosis in L-02 hepatocyte to establish NAFLD cell model in vitro and explore the protective effect of resveratrol on steatosis cell model of L-02 cells.Methods: In this study,human L-02 normal hepatocytes were stimulated with different concentrations of free fatty acids to induce hepatocyte steatosis.The effects of different concentrations of FFA on L-02 hepatocytes were detected by CCK8,oil red O,flow cytometry and other methods.The effect of different doses of resveratrol on the proliferation activity of L-02 hepatic steatosis cell model was detected by CCK8.Results: The CCK8 assays showed that the proliferative activity of cells gradually weakened along with the increase of FFA concentration in a certain range.In addition,the cell proliferation activity gradually weakened with the prolongation of FFA stimulation within 24 hours.When the L-02 liver cells stimulated with the high-FFA stained with oil red O,it shown that a large number of fat granules and vesicular fat lesions appeared in the cytoplasm,the steatosis was significant,and the hepatocytes were atrophic and varied in size compared with the normal L-02 liver cells.In the L-02 hepatocytes stimulated by low concentration of FFA,fat vacuoles were observed occasionally and steatosis was not obvious.The flow cytometry results showed that the early apoptosis rate of L-02 hepatocytes stimulated by high concentration of FFA increased significantly and the proapoptosis effect of FFA was in a dose-dependent manner.We further observed that different concentrations of RSV improved the proliferation activity of L-02 hepatocytes stimulated by 0.75% FFA at 48 hours.Conclusions: This study demonstrated that the L-02 hepatic steatosis cell model was successfully established through stimulation of FFA.This model provides a more convenient experimental method for NAFLD therapeutic research.Resveratrol has a significant protective effect on the proliferation of L-02 hepatic steatosis cell model,and the underlying mechanism needs further study.