Effects Of Uric Acid On Steatosis Hepatocytes Oxidative Stress And Mitochondrion Function

Objective: To research the possible antioxidative and pro-oxidative effect of different concentration uric acid(UA)with different intervened time on steatosis hepatocytes which induced oxidative stress(OS)and to explore the possible mechanism of UA in pro-or antioxidative process.Method: L-02 cells were intervened via 0.1,0.2,0.3 and 0.4 mmol/L OA,cell viability and TG content detection were used to confirm the best OA concentration of steatosis model.5,10,20 and 30 mg/d L UA were used to intervene the model for 24,48,72 and 96 h respectively,normal cell and model cell set as control.The intervening time we chosen were 24,48,72 and 96 h.The fluorescence intensity of ROS,total SOD activity,GSH and MDA content were detected according to corresponding kits;the transcription and translation level of SOD1 and ?-GCLC were detected via RT-qPCR,Western blot and ICC;AST,ALT SDH,CCO and ATP synthase activities,apoptosis level,and the content of 8-OHdG were detected by corresponding kits;the ultrastructure of the cells and mitochondrion were observed by electron microscope at last.Results: 1.3 mmol/L group,which did not have statistic difference with other groups on cell viability and contain most TG,was chose as best modeling concentration.2.ROS intensity,MDA content and total SOD activity in 5 and 10 mg/dL groups were decreased and the GSH content increased compare with model group(P<0.05).3.The SOD1 trancription level in 5,10 and 30 mg/d L group obviously lower than model group(P<0.05).The translation level of SOD1 in 5 mg/d L group was higher than model group at 24 and 48 h as well as in 10 mg/d L which higher than model group at 72 and 96 h.The transcription level of ?-GCLC in 5 and 10 mg/dL group were lower than model group at 24 h and 48 h(P<0.05).The translation level of ?-GCLC in model group was lower than normal group at 24 h,48 h and 72 h(P<0.05),and,the level in 5 mg/d L group was higher than model group at 72 h as well as 10 mg/d L group at 48 h,72 h and 96 h.4.ALT activity in 5 mg/d L group was lower than model group(P<0.05)and the apoptosis rate in 5 and 10 mg/dL group decreased either.5.5 and 10 mg/d L UA decreased the content of 8-OH-d G(P<0.05).6.5 and 10 mg/dL UA in immunocellulerchemistry research of SOD1 and ?-GCLC increased the quantity of mitochondria,especially in 10 mg/d L group,the lipid droplet could not be observed no more.Conclusion: 5 and 10 mg/d L UA may provide a protect effect in steatosis cell,and it owe possible to the antioxidative capacity of UA.However,30 mg/dL UA could provid limited protection within 24 h,it would promote OS after 72 h.