银杏叶提取物对H_2O_2诱导的巨噬细胞凋亡的抑制作用

Suppressive Effects of Ginkgo biloba Extract on H_2O_2-Induced Apoptosis in RAW264.7 Macrophages

目的研究银杏叶提取物(EGb761)对H_2O_2所致RAW264.7巨噬细胞凋亡的保护作用。方法以H_2O_2诱导RAW264.7巨噬细胞凋亡为实验模型,用噻唑蓝(MTT)实验、流式细胞术、蛋白质印迹分析、琼脂糖凝胶电泳和荧光分析分别检测细胞存活率、线粒体膜电位、细胞色素C的释放和Bax,bcl-2蛋白水平、DNA的降解、半胱氨酰天冬氨酸特异性蛋白酶-3(caspase-3)活性。结果EGb761能明显降低H_2O_2对RAW264.7细胞的氧化损伤,提高细胞的存活率；维持线粒体膜的完整性,抑制跨膜电位的耗散和细胞色素C的释放；抑制caspase-3的活化和DNA的降解。结论EGb761具有清除活性氧,减轻H_2O_2所致RAW264.7细胞的氧化损伤,在对H_2O_2诱导的细胞凋亡中发挥重要的抗凋亡作用。

OBJECTIVE To study protective effects of Ginkgo biloba extract （EGb761） on hydrogen peroxide （H2O2）- induced cell apoptosis in RAW264.7 macrophages.METHODS ：aoratorial model of hydrogen peroxide-induced apoptosis in RAW264.7 macrophages was established. The growth activities of the RAW264.7 cells were measured by MTT assay. The low cytometery （FCM） was used to detect mitochondrial membrane potential（ △ψm）. The release of cytochrome C, bcl-2 and Bax protein level were measued by Western blotting. The DNA fragmentation was examined by agarose gel electrophoresis. The fluorescent analysis was used to detect the activity of caspase-3. RESULTS The results showed that EGb761 evidently decreased H2O2-induced oxidative damage in RAW264.7 macrophages, and improved cell viability, and maintained the integrality of mitochondrial membrane, and inhibited the debaser of mitochondrial membrane potential and release of cytochrome C, and inhibited the activity of caspase-3 and DNA fragmentation. CONCLUSION EGb761 could clean out reactive oxygen species （ROS） and attenuate oxidative damage induced by hydrogen peroxide in RAW264.7 cells. EGb761 might own the resistance of cell apoptosis induced by H2O2.