2018 年第 11 期 第 13 卷

白英生物碱对叔丁基过氧化氢诱导的人神经瘤母细胞系SH-SY5Y细胞氧化应激和凋亡的作用机制研究

Study on the mechanisms of solanum lyratum thunberg alkaloid against tert-butyl hydroperoxide-induced oxidative stress and apoptosis in human neuroma metrocyte cell line SH-SY5Y cells

作者：霍焰王淳王梅王晓波韩兆丰董秀

英文作者：

单位：110034沈阳，辽宁中医药大学附属第二医院心病二科［霍焰（辽宁中医药大学基础医学专业在职硕士研究生）］；110847沈阳，辽宁中医药大学基础医学院（王淳、王梅、王晓波、韩兆丰、董秀）

英文单位：

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

英文关键词：

• 摘要：

• 【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（100 μmol/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100 μmol/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）。20 mg/L白英生物碱和100 μmol/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。

• 【Abstract】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.