The Study On The Therapeutic Effect Of Ursolic Acid On Non-alcoholic Fatty Liver Disease

Objective:In this paper,insulin and oleic acid induced Hep G2 cells to establish insulin-resistant NAFLD cell model in order to study the impact of ursolic acid on non-alcoholic fatty liver disease.Methods: Human Hep G2 cells were cultured in high oleic acid medium supplemented with insulin to establish an in vitro cell model of insulin resistance NAFLD.The model group was given different concentrations of ursolic acid intervention,cell proliferation assay by CCK-8 method,screening the best concentration of the drug.The experiment was randomly divided into blank + insulin control group,oleic acid + insulin model group,oleic acid + insulin + ursolic acid experimental group,oleic acid + insulin + metformin control group.The effects of UA on the survival of fatty liver cells were detected by CCK-8 assay.The morphology and intracellular lipid droplets formation in different groups were observed by oil red O staining.The changes of TG content in liver cells were detected by TG kit.Result:(1)Hep G2 cells were cultured in DMEM high glucose medium supplemented with 20 mg/ml oleic acid and 50 μIU/ml insulin for 48 h,and an insulin resistant NAFLD cell model was successfully established.After oil red O staining,a large number of orange-colored round lipid droplets formed around the inner nucleus of the cell were observed under an inverted microscope.The cell morphology was normal,the nuclei were clear,and the cell activity was good.The TG content in the normal control group was 1.05 mmol/g,and the TG content in the model group was 3.62mmol/g,which was significantly higher than that in the control group(P<0.05).(2)5,10,and 20 μmol/L ursolic acid were added to the model group,and the optimal concentration of the drug was determined by measuring the cell proliferation rate by CCK-8 after 48 hours of culture.Finally,5μmol/L ursolic acid was selected as the optimal concentration of the drug.The cell proliferation rate was about 100%.With the increase of ursolic acid concentration,the cell proliferation rate gradually decreased.(3)In the model group,the intracellular TG content was 3.41 mmol/g,and the intracellular TG content after treatment with 5 μmol/L ursolic acid for 48 h was 1.19mmol/g,which was significantly lower than that in the model group(P<0.05).Conclusion:Human Hep G2 cells were cultured in DMEM high-glucose medium supplemented with 50 μIU/ml insulin and 20 mg/ml oleic acid for 48 hours to establish a successful insulin resistance NAFLD model.5 μmol/L is the optimal concentration of ursolic acid for the fatty liver cell model.Ursolic acid has the effect of reducing TG content in fatty liver cells,so ursolic acid has the effect of treating NAFLD.