银杏叶提取物对百草枯致PC12细胞损伤的保护作用及机制

Neuroprotective effects of ginkgo biloba extract against paraquat-induced PC12 cell line damage and its mechanism

目的探讨百草枯(PQ)对PC12细胞的损伤作用,观测银杏叶提取物(EGB761)对该损伤的保护作用并探讨其作用机制.方法MTT比色试验检测细胞活性;乳酸脱氢酶(LDH)测定细胞损伤程度;流式细胞仪(FCM)检测细胞凋亡;caspase-3免疫细胞化学染色观察细胞凋亡.结果40mg/L EGB761对300μmol/LPQ作用于PC12细胞24h,具有一定的保护作用;EGB761可明显抑制PQ诱导的PC12细胞LDH的释放(0.25±0.01)(与PQ组比较,P<0.05);EGB761可降低PC12细胞的凋亡百分率,PQ组凋亡百分率为66.7%,使用EGB761保护后凋亡百分率降为17.1%;并可减少PQ诱导的PC12细胞caspase-3过度表达.结论EGB761可减轻PQ诱导的PC12细胞损伤和凋亡,该作用可能与抑制caspase-3蛋白酶的激活有关.

AIM： To study the neuroprotective effect and related mechanisms of ginkgo biloba extract （ EGB761 ） against paraquat- （PQ） induced apoptosis in pheochromocytoma （PC12） cells. METHODS： Using PQ-induced PC12 cells damage as the model of Parkinson＇s disease in vitro, the cytotoxicity of PQ was measured by 3-[4, 5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide （MTT） assay, the impairment of PC12 cells was detected by lactate dehydrogenase （LDH） assay, the apoptosis of PC12 cells was detected by flow cytometry and the morphological change of PC12 cells was observed by caspase-3 immunocytochemical staining. RESULTS： 40 mg/L EGB761 played a protective role in the damage induced by 300 μmol/L PQ for 24 h. EGB761 significantly inhibited LDH release induced by PQ in PC12 cells （0.25 ＋0.01） （P 〈0. 05 vs PQ group）. EGB761 decreased the rate of apoptosis to 17. 1% and the overexpression of caspase-3 induced by PQ in PC12 cells. CONCLUSION： EGB761 can protect cells from the necrosis and apoptosis induced by PQ, which may be related to the inhibition of caspase-3 protease activation.