Effects of ursolic acid on hydrogen peroxide-induced inflammatory response and oxidative stress in nerve cells PC12

HU Canying; DAI Xiaodan; HUANG Xi

doi:10.13431/j.cnki.immunol.j.20240114

Abstract：Objective To investigate the effect of ursolic acid on the inflammatory response and oxidative stress induced by hydrogen peroxide(H2O2)in nerve cells PC12 and its related mechanisms.Methods Nerve cells PC12 were treated with H2O2 to establish a cell injury model,and then treated with various concentrations of ursolic acid(2.5,5,10,and 20 μmol/L).MTT experiment was used to detect cell viability.PC12 cells were divided into control group(Con),hydrogen peroxide group(H2O2),hydrogen peroxide+ursolic acid group(H2O2+ursolic acid).Flow cytometry was used to detect cell apoptosis;Western blot was used to detect the expression of Cleaved cas3,Bax,Bcl-2,NF-κB,iNOS,NO,mTOR pathway proteins;DCFH-DA method was used to detect the relative level of ROS,while ELISA was used to detect oxidative stress and inflammatory response indicators.Results Compared with the Con group,the cell viability of the H2O2 group was significantly reduced;compared with the H2O2 group,the cell viability of the H2O2+2.5,5,10,20 μmol/L ursolic acid groups increased.Compared with the Con group,the apoptosis rate,LDH content,relative level of ROS,TNF-α,IL-1β in H2O2 group were significantly increased,CAT activity and SOD activity were significantly decreased,Cleaved cas3,Bax,NF-κB,iNOS,NO,P-mTOR,p-p70S6K protein expression increased,Bcl-2 protein expression decreased.Compared with the H2O2 group,the apoptosis rate,LDH content,relative level of ROS,TNF-α,IL-1β in the H2O2+ursolic acid groups were significantly decreased,CAT activity and SOD activity were significantly increased,Cleaved cas3,Bax,NF-κB,the expression of iNOS,NO,p-mTOR,and p-p70S6K protein decreased,and the expression of Bcl-2 protein increased.Conclusion Ursolic acid may reduce the apoptosis,inflammation and oxidative stress induced by hydrogen peroxide in neuronal PC12 cells by regulating the mTOR signaling pathway.