叶黄素抑制叔丁基过氧化氢诱导的视网膜神经节细胞凋亡  

Lutein suppresses t-BHP-induced apoptosis in retinal ganglion cells

作　　者：张婵娟[1,2] 赵佳仪 黄翠芹[1] 李勤[1] 王珍[1] 甘丹卉[1] 朱丽红[1] 陆大祥[1] 

机构地区：[1]暨南大学医学院病理生理学系,国家中药管理局三级科研实验室,广东广州510632 [2]安徽人口职业学院护理系,安徽池州247099

出　　处：《中国病理生理杂志》2016年第6期1043-1050,共8页Chinese Journal of Pathophysiology

基　　金：国家重点基础研究发展计划(973计划)(No.2011CB707501);国家自然科学基金资助项目(No.81371442);广州市科技计划项目重大专项(No.11Bpp ZXaa2070006)

摘　　要：目的:观察叶黄素(lutein)对叔丁基过氧化氢(t-BHP)处理的视网膜神经节细胞(RGC-5细胞系)的保护效应并探讨其作用机制。方法:用免疫荧光染色检测视网膜神经节特异性蛋白Brn-3和神经微管结合蛋白MAP-2的表达来鉴定RGC-5细胞;将RGC-5细胞随机分为对照组、t-BHP处理组、t-BHP和lutein共同处理组、lutein处理组,培养24 h,MTT实验检测细胞活力;Annexin V-FITC/PI双染流式细胞术检测细胞凋亡;免疫细胞化学技术检测caspase-3蛋白的活化情况;Western blot检测Bcl-2/Bax、cleaved caspase-3、JNK和c-Jun蛋白的变化。结果:MTT实验和流式细胞检测结果显示,lutein能提高t-BHP处理的RGC-5细胞的活力,并降低t-BHP诱导的RGC-5细胞凋亡;免疫荧光结果显示lutein能抑制t-BHP诱导的caspase-3的活化;与对照组比较,t-BHP处理后RGC-5细胞抗凋亡蛋白Bcl-2表达下调(P<0.05),Bax/Bcl-2比率升高,cleaved caspase-3表达上调(P<0.05),JNK和c-Jun蛋白的磷酸化水平增加(P<0.05),t-BHP的上述作用可被lutein部分逆转。结论:Lutein能够降低t-BHP诱导的RGC-5细胞凋亡,其机制与其上调Bcl-2的表达、抑制caspase-3的活化并降低JNK和c-Jun蛋白的磷酸化有关。AIM： To investigate the protective effects of lutein on retinal ganglion cells in vitro. METHODS：The effect of lutein on tert-butyl hydroperoxide（ t-BHP）-treated retinal ganglion cells（ RGC-5 cell line） was determined.The protein expression of Brn-3 and MAP-2 was examined by the method of immunocytochemistry to identify the RGC-5cells. The RGC-5 cells were induced by a 24 h exposure of t-BHP,and the cell viability was examined by MTT assay. The apoptotic ratio of the RGC-5 cells after exposed to t-BHP or / and lutein treatment was analyzed by flow cytometry assay with Annexin V-FITC / PI staining. The activation of caspase-3 was detected by immunocytochemistry and the protein levels of Bcl-2,Bax,cleaved caspase-3,JNK and c-Jun were determined by Western blot. RESULTS： The RGC-5 cells expressed Brn-3 and MAP-2 proteins. Lutein treatment prevented t-BHP-induced RGC-5 cell apoptosis and increased the cell activity.Compared with control group,exposure of the RGC-5 cells to t-BHP decreased the expression of anti-apoptotic protein Bcl-2,increased the Bax / Bcl-2 ratio,up-regulated the level of cleaved caspase-3,also promoted the phosphorylation of JNK and c-Jun. Lutein partly reversed the effects of t-BHP on the RGC-5 cells mentioned above. CONCLUSION： Lutein protects RGC-5 cells against t-BHP-induced apoptosis by up-regulating Bcl-2 expression and inhibiting caspase-3 activation through modulating the JNK signaling pathway.

关 键 词：叔丁基过氧化氢 叶黄素 RGC-5细胞系 细胞凋亡 

分 类 号：R329.21[医药卫生—人体解剖和组织胚胎学]