银杏叶提取物对百草枯致PC细胞损伤的保护作用及机制.docxVIP

银杏叶提取物对百草枯致PC12细胞损伤的保护作用及机制 作者：康小刚，陈建宗，王百忍，徐震，李海龙，奚淑芳 【关键词】 银杏叶提取物 Neuroprotective effects of ginkgo biloba extract against paraquatinduced PC12 cell line damage and its mechanism 【Abstract】 AIM: To study the neuroprotective effect and related mechanisms of ginkgo biloba extract against paraquat induced apoptosis in pheochromocytoma cells. METHODS: Using PQinduced PC12 cells damage as the model of Parkinsons disease in vitro, the cytotoxicity of PQ was measured by 3［4, 5dimethylthiazol2yl］2, 5diphenyltetrazolium bromide assay, the impairment of PC12 cells was detected by lactate dehydrogenase assay, the apoptosis of PC12 cells was detected by flow cytometry and the morphological change of PC12 cells was observed by caspase3 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 . EGB761 decreased the rate of apoptosis to % and the overexpression of caspase3 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 caspase3 protease activation. 【Keywords】 ginkgo biloba extract; PC12 cells; paraquat; apoptosis; neuroprotection 【摘要】 目的： 探讨百草枯对PC12细胞的损伤作用，观测银杏叶提取物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；乳酸脱氢酶测定细胞损伤程度；流式细胞仪检测细胞凋亡；caspase3免疫细胞化学染色观察细胞凋亡. 结果： 40 mg/L EGB761对300 μmol/L PQ作用于PC12细胞24 h，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；EGB761可降低PC12细胞的凋亡百分率，PQ组凋亡百分率为%，使用EGB761保护后凋亡百分率降为%；并可减少PQ诱导的PC12细胞caspase3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caspase3蛋白酶的激活有关. 【关键词】银杏叶提取物；PC12细胞；百草枯；细胞凋亡；神经保护 0引言 流行病学资料显示，环境因素如除草剂、杀虫剂等与帕金森病的发生有关［1］. 二氢吡啶类除草剂百草枯是一种常用农药，其结构与神经毒素MPTP的活性代谢产物MPP+极其相似，也具有与MPP+相类似的神经毒性［2］. 我们拟用其作为建立帕金森病体外模型的工具药物，研究银杏叶提取物对PQ诱导的PC12细胞毒性损伤的保护作用及可能的作用机制. 　1材料和方法 材料 EGB761为德国威玛舒培博士药厂产品；PQ、胰蛋白酶、4甲基偶氮唑蓝, Annexin VFITC, PI为Si