银杏内酯B对谷氨酸诱导SH-SY5Y细胞凋亡的保护作用  

The protective effects of Ginkgolide B on glutamic acid induced apoptosis in SH-SY5Y cells

作　　者：宋颖[1] 李尘远[2] 张孟姝[1] 吴遥[1] 宋长威[1] 梅晰凡[2] 

机构地区：[1]辽宁医学院附属第一医院,锦州121000 [2]辽宁医学院,锦州121000

出　　处：《中药药理与临床》2015年第5期23-26,共4页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金(81171799/H1504)

摘　　要：目的:探讨银杏内酯B对谷氨酸诱导的人神经母细胞瘤(SH-SY5Y)细胞凋亡的保护作用及其机制。方法:用2mmol/L谷氨酸诱导SH-SY5Y细胞凋亡建立模型,以不同浓度的银杏内酯B(10、50和250μmol/L)进行保护,MTT比色法检测细胞存活率;Hoechst 33258染色法观察药物作用后细胞凋亡的形态学变化;流式细胞仪检测细胞凋亡率;Fura-2/AM双波长荧光法测定细胞内钙离子浓度;Western blot检测线粒体和胞浆中Cyt C蛋白的表达。结果:2 mmol/L谷氨酸作用SH-SY5Y细胞24 h后细胞存活率为0.721±0.013,与正常组相比明显降低;细胞核固缩裂解,呈凋亡特征性改变;细胞凋亡率为(13.27±0.08)%、细胞内钙离子浓度为359±11 nmol/L,均明显升高。与凋亡组相比,银杏内酯B(10、50、250)μmol/L保护组细胞存活率分别为0.768±0.014、0.819±0.013、0.897±0.010,均明显升高;细胞核核固缩裂解减少,形态有所改善;细胞凋亡率分别为11.33±0.57、7.02±0.47、5.85±0.24;细胞内钙离子浓度为302±9 nmol/L、243±9 nmol/L、186±8 nmol/L,均明显降低;Western blot结果显示,与凋亡组相比银杏内酯B保护组可以显著减少细胞线粒体以及胞浆中Cyt C蛋白的含量。结论:银杏内酯B对谷氨酸诱导的SH-SY5Y细胞凋亡具有保护作用,其机制可能与银杏内酯B降低了细胞内钙离子的浓度及阻断了Cyt C从线粒体释放入胞浆中有关。Objective： To investigate the protective effects and mechanism of Ginkgolide B on apoptostic SH-SY5 Y cells induced by glutamic acid.Methods： The SH-SY5 Y cells were used as models,the apoptosis were induced by glutamic acid 5 mmol / l. Different concentration of Ginkgolide B（ 10,50,250） μmol / L were used to protect the cells. The cell viability was analysed by MTT assay; the changes of nucleus were assayed by Hoechst 33258 staining; the apoptosis rate was measured by flow cytometry using Annexin-V and propidumiodide（ PI）; the fluorescence intensity of intracellular calcium was measured by Fura-2 / AM double wavelength fluorescence. Expression of Cyt C protein in mitochondria and cytoplasm of Western blot. Results： Compared with normal control group,the cell viability of apoptosis group was significantly decreased to 0. 721 ± 0. 013 after 24 h（ P〈0. 01）; the karyopyknosis and split showed cells apoptosis; the apoptosis rate and the fluorescence intensity of intracellular calcium increased to 13. 27 ± 0. 08% and 359 ± 11 nmol / L,respectively（ P〈0. 05; P〈0. 01）.Compared with apoptosis group,the cell viability in Ginkgolide B（ 10,50,250） μmol / L protect groups were 0. 768 ± 0. 014,0. 819 ± 0. 013 and 0. 897 ± 0. 010,respectively（ P〈0. 05; P〈0. 01）; karyopyknosis and split decreased and the morphologies of cells significantly improved; the apoptosis rate and the fluorescence intensity of intracellular calcium were 11. 33 ± 0. 57,7. 02 ± 0. 47,5. 85 ± 0. 24 and 302 ± 9nmol / L,243 ± 9 nmol / L,186 ± 8 nmol / L（ P〈0. 05; P〈0. 01）. Western Blot showed that the protective group of Ginkgolide B could significantly decrease the contents of Cyt C in the cell mitochondria and cytoplasm of the cells in the group of the apoptosis group（ P〈0. 05）.Conclusion： Ginkgolide B on glutamate induced apoptosis of SH-SY5 Y cells has a protective effect,the mechanism may be related to the effects of Ginkgolide B reduces the intracellular calcium ion concentration and

关 键 词：银杏内酯B 谷氨酸 SH-SY5Y细胞 细胞凋亡 钙离子浓度 细胞色素C 

分 类 号：R285[医药卫生—中药学]