The Effect Of Autophagy To PC12 Cells' Toxicity Which Induced By H₂O₂

Objective:To observe the effects of autophagy inhibitor 3-MA to PC 12 cells' apoptosis and explore its possible mechanism.Method:(1)The ability of PC12 cells were detected by MTT assay,cell culture supernatant LDH activity assay and fluorescent staining.(2)Mitochondrial transmembrane potential was detected by Rh123.(3)The intracellular reactive oxygen species level(ROS),intracellular content of malondialdehyde(MDA),the activity of superoxide dismutase(SOD)and peroxidase(CAT)were detected.(4)Autophagy was detected by staining of MDC.(5)The change of NF-E2-related factor(Nrf2)and LC3-?/LC3-? in PC 12 cells were analysed by Western blot experiments analysis.Result:The first part:It was found that PC12 cells were damaged in a concentration-dependent manner after H2O2 treatment.And 200?mol/L,400?mol/L,800?mol/L H2O2 significantly induced PC 12 apoptosis,and decline mitochondrial transmembrane potential.The second part:The level of ROS and lipid peroxidation in PC12 cells significantly increased,and the antioxidant enzyme activities including SOD,CAT and the expression of Nrf2 decreased after H2O2.The third part:Compared with the group which damaged by 400?mol/L H2O2 for 12 hours,MDC punctate structures and the level of LC3-?/LC3 of the group for 24 hours increased significantly.The forth part:Compared with the group with 400?mol/L H2O2,the apoptosis,the level of ROS and lipid peroxidation of group with 3-MA and 400?mol/L H2O2 simultaneously significantly increased,and mitochondrial transmembrane potential,antioxidant enzyme activities including SOD,CAT,and the expression of Nrf2 decreased.Conlusion:Hydrogen peroxide induced PC12 cells apoptosis,3-MA promoted hydrogen peroxide-apoptosis through inhibiting autophagy,its mechanism may be related to the expression of intracellular Nrf2.