摘要
目的探讨白英生物碱对叔丁基过氧化氢(tBHP)诱导的人神经瘤母细胞系SH-SY5Y细胞氧化应激和细胞凋亡的作用。方法将人神经瘤母细胞系SH-SY5Y细胞分为对照组、tBHP模型组、白英生物碱(10、20、40 mg/L)组和维生素E(100μmol/L)组。采用甲基噻唑蓝法检测细胞活力,倒置显微镜观察细胞形态学变化,TUNEL法检测细胞凋亡,流式分析法检测线粒体膜电位(罗丹明123荧光强度)和活性氧水平[二氯荧光素(DCF)荧光强度],蛋白质印迹法检测细胞凋亡相关蛋白Bcl-2和Bax的表达。结果 tBHP模型组细胞存活率和罗丹明123荧光强度明显低于对照组,细胞凋亡率和DCF荧光强度明显高于对照组;10、20、40 mg/L白英生物碱组和100μmol/L维生素E组细胞存活率和罗丹明123荧光强度明显高于tBHP模型组[(48±18)%、(61±3)%、(74±19)%、(86±35)%比(43±3)%;(60. 84±0. 11)%、(71.32±0.24)%、(79.68±0.15)%、(70.47±0. 32)%比(55.43±0. 23)%],细胞凋亡率和DCF荧光强度明显低于tBHP模型组[(31.11±0.09)%、(18.34±0.21)%、(13. 58±0. 51)%、(22. 82±0. 39)%比(37. 16±0.27)%;(62.34±0.53)%、(41.22±0.44)%、(38.57±0.65)%、(49.18±0.52)%比(75.42±0.46)%],差异均有统计学意义(均P<0.05)。20mg/L白英生物碱和100μmol/L维生素E能增加tBHP诱导的SH-SY5Y细胞中Bcl-2蛋白表达,同时抑制Bax蛋白表达。结论白英生物碱能够抑制SH-SY5Y细胞中活性氧产生,降低tBHP诱导的SH-SY5Y细胞的氧化应激,减少细胞凋亡。
Objective To investigate the mechanisms of solanum lyratum thunberg alkaloid(STA) against tert-butyl hydroperoxide(tBHP)-induced oxidative stress and apoptosis in human neuroma metrocyte cell line SH-SY5Y cells in vitro. Methods Human neuroma metrocyte cell line SH-SY5Y cells were divided into control group, tBHP model group, STA(10, 20, 40 mg/L) group and vitamin E(100 mol/L) group. Cell viability was measured by MTT assay. Cell morphology was observed by light microscope. Cell apoptosis was detected by TUNEL assay. Mitochondrial membrane potential[fluorescence intensity of rhodamine-123(Rh123)] and reactive oxygen species(ROS)[fluorescence intensity of dichlorofluorescein(DCF)] were detected by flow cytometric assay. Expressions of Bax and Bcl-2 protein were tested by western blotting. Results Cell survival rate and fluorescence intensity of Rh123 in tBHP model group were significantly lower than those in control group; apoptosis rate and fluorescence intensity of DCF in tBHP model group were significantly higher than those in control group; cell survival rate and fluorescence intensity of Rh123 in 10, 20, 40 mg/L STA groups and vitamin E group were significantly higher than those in tBHP model group[(48±18)%,(61±3)%,(74±19)%,(86±35)% vs (43±3)%; (60.84±0.11)%,(71.32±0.24)%,(79.68±0.15)%,(70.47±0.32)% vs (55.43±0.23)%]; apoptosis rate and fluorescence intensity of DCF in 10, 20, 40 mg/L STA groups and vitamin E group were significantly lower than those in tBHP model group[(31.11±0.09)%,(18.34±0.21)%,(13.58±0.51)%,(22.82±0.39)% vs (37.16±0.27)%; (62.34±0.53)%,(41.22±0.44)%,(38.57±0.65)%,(49.18±0.52)% vs (75.42±0.46)%](all P〈0.05). Western blotting results indicated that 20 mg/L STA and 100 μmol/L vitamin E could increase the expression of BCL-2 protein and inhibit the expression of BAX protein in tBHP-induced SH-SY5Y cells. Conclusion STA can significantly inhibit tBHP-induced ROS production, oxidative stress and apoptosis in SH-SY5Y cells.
作者
霍焰
王淳
王梅
王晓波
韩兆丰
董秀
Huo Yan;Wang Chun;Wang Mei;Wang Xiaobo;Han Zhaofeng;Dong Xiu(The Second Department of Heart Disease,the Second Affiliated Hospital of Liaoning University of Traditional Chinese Medicine,Shenyang 110034,China;College of Basic Medicine,Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China)
出处
《中国医药》
2018年第11期1738-1741,共4页
China Medicine
基金
辽宁省科学技术计划(201501094)
辽宁省教育厅科学技术研究项目(L201603)
沈阳市科技计划(F16-205-1-20)~~
关键词
SH-SY5Y细胞
白英生物碱
叔丁基过氧化氢
细胞凋亡
活性氧
氧化应激
SH-SY5Y cells
Solanum lyratum thunberg alkaloid
Tert-butyl hydroperoxide
Apoptosis
Reactive oxygen species
Oxidative stress