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

发布时间：2018-03-03 08:21

本文选题：叔丁基过氧化氢　切入点：叶黄素　出处：《中国病理生理杂志》2016年06期 　论文类型：期刊论文

【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(P0.05),Bax/Bcl-2比率升高,cleaved caspase-3表达上调(P0.05),JNK和c-Jun蛋白的磷酸化水平增加(P0.05),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。
[Abstract]:Objective: To observe the effect of lutein (Lutein) of tert butyl hydroperoxide (t-BHP) treatment of retinal ganglion cells (RGC-5 cell line) the protective effect and explore its mechanism. Methods: to identify the RGC-5 cell binding protein MAP-2 expression by immunofluorescence staining of retinal ganglion specific protein Brn-3 and microtubule RGC-5 cells were divided; for the control group, t-BHP treatment group, t-BHP and lutein treatment group, lutein treatment group, cultured for 24 h, the cell viability by MTT assay; Annexin V-FITC/PI double staining flow cytometry; activation of caspase-3 protein was detected by immunocytochemical technique; Western blot Bcl-2/Bax cleaved Caspase-3, change detection, JNK and c-Jun protein. Results: MTT assay and flow cytometry assay showed that lutein can improve the t-BHP treated RGC-5 cell viability and reduced apoptosis induced by low t-BHP RGC-5 Apoptosis; immunofluorescence results showed that activation of lutein can inhibit t-BHP induced caspase-3; compared with the control group, Bcl-2 protein expression of RGC-5 cell apoptosis after t-BHP treatment (P0.05), Bax/Bcl-2 ratio increased, cleaved expression of Caspase-3 (P0.05), JNK and c-Jun protein phosphorylation level increased (P0.05), the role of t-BHP can be partially reversed lutein. Conclusion: Lutein can reduce the RGC-5 induced apoptosis of t-BHP cells, its mechanism is associated with upregulation of Bcl-2 expression, inhibited the activation of Caspase-3 and JNK and decrease the phosphorylation of c-Jun protein.

【作者单位】：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;安徽人口职业学院护理系;
【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)
【分类号】：R774.1

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