The Protective Effect Of Trihexylphenidyl On H₂O₂-induced Injury In PC12 Cells

Objective:To investigate the protective effect of trihexylphenidyl?THY?on H₂O₂-induced cell injury and its mechanism in rat pheochromocytoma cells?PC12?.Methods:H₂O₂ was used to induce the injury in PC12 cells and construct the oxidative damage model.The cell viability of THY on PC12 cells and the protective effect of THY on H₂O₂-induced cell injury was detected by MTT assay.Then the morphology of PC12 cells in different situations was observed by fluorescence microscope.DNA-binding dye AO/EB and PI/Hoechst33342 were used to observe the morphology of cell apoptosis.Annexin V-FITC/PI kit was used to detect the apoptosis rate of PC12 cells.The expression of Nrf2 in H₂O₂-induced PC12 cells was detected by Immunostaining and Western Blot.the protein expression of p-Akt,p-ERK1/2 and HO-1 were detected by Western Blot.Results:1.The cell viability of PC12 cells treated with H₂O₂ was significantly decreased.After pretreatment with THY and NAC for 2 hours,the survival rate of PC12 cells was obviously increased.2.Different concentrations of trihexylphenidyl?0.01?0.1?1.0?M THY?could reduce the apoptosis rate of H₂O₂-induced PC12 cells injury and significantly improve the morphology of damaged PC12 cells effectively.3.THY can upregulate the expression of p-ERK1/2,p-Akt and HO-1 in H₂O₂-induced PC12cells.4.After pretreatment with THY,the level of Nrf2 was significantly increased and Nrf2 was activated to induce nuclear translocation in H₂O₂-induced PC12 cells.However,the addition of inhibitor LY294002and PD98059 significantly decreased the level of Nrf2.Conclusions:THY can protect H₂O₂-induced oxidative damage in PC12 cells,and this protective effect may be related to PI3K/Akt and ERK signaling pathways regulating Nrf2 nuclear translocation,thereby activating downstream target genes and inducing the expression of antioxidant proteins?such as HO-1?.