雷帕霉素对小鼠星形胶质细胞体外诱导凋亡的影响

发布时间：2018-03-20 03:30

本文选题：雷帕霉素　切入点：星形胶质细胞　出处：《中国病理生理杂志》2015年04期 　论文类型：期刊论文

【摘要】：目的:探讨雷帕霉素(rapamycin,Rapa)对小鼠星形胶质细胞体外凋亡的影响。方法:无菌分离并体外培养C57BL/6J幼鼠脑组织星形胶质细胞。通过MTT比色法测定并分析Rapa浓度对幼鼠星形胶质细胞存活的影响;SYTOX屮Green荧光染色联合荧光酶标仪检测并分析Rapa对H2O2、ionomycin、deferorxamine等诱导剂作用一定时间内细胞存活的影响;Di OC6(3)染色分析Rapa在H2O2氧化应激损伤条件下对星形胶质细胞线粒体膜电势的影响;分别采用H2DCFDA和Mito SOXTMRed荧光染色联合流式细胞术检测Rapa预适应对星形胶质细胞ROS生成以及线粒体内ROS含量的影响。结果:Rapa能促进H2O2以及ionomycin联合deferorxamine损伤作用下的星形胶质细胞的存活,对线粒体膜电势有保护作用,可降低H2O2损伤作用下星形胶质细胞ROS的产生并可以维持胞内线粒体ROS的含量在较低水平。结论:Rapa能够减少细胞内ROS的生成量并降低胞内线粒体内ROS水平;能够减轻H2O2对细胞线粒体膜的损伤破坏,维护线粒体膜电势的稳定性,进而对氧化应激损伤介导细胞凋亡有一定的抑制作用。
[Abstract]:Objective: to investigate the effect of rapamycin rapamycin Rapa on apoptosis of mouse astrocytes in vitro. Methods: C57BL / 6J young rat brain astrocytes were isolated and cultured in vitro. The concentration of Rapa was determined and analyzed by MTT colorimetry. The effect of Rapa on Cell Survival in a certain period of time the effect of Rapa on Cell Survival of H _ 2O _ 2H _ 2O _ 2 induced by ionomycinycinycin deferorxamine and other inducers was analyzed by Rapa staining under oxidative stress of H _ 2O _ 2. The effect of mitochondrial membrane potential on glial cells; The effects of Rapa preconditioning on ROS production and ROS content in mitochondria of astrocytes were detected by H _ 2DC FDA and Mito SOXTMRed fluorescence staining combined with flow cytometry. Results: h _ 2O Rapa could promote the survival of astrocytes induced by H _ 2O _ 2 and ionomycin combined with deferorxamine damage. It has protective effect on mitochondrial membrane potential. It can reduce the production of ROS and maintain the low level of mitochondrial ROS in astrocytes after H _ 2O _ 2.Conclusion:: Rapa can reduce the production of ROS and decrease the level of ROS in mitochondria. It can attenuate the damage of mitochondria membrane induced by H2O2, maintain the stability of mitochondrial membrane potential, and inhibit apoptosis induced by oxidative stress.
【作者单位】：暨南大学附属第一医院临床检验中心;暨南大学生命科学技术学院组织移植与免疫研究中心;
【基金】：国家自然科学基金资助项目(No.31200667) 暨南大学科研培育与创新基金青年基金(理工医类)资助项目(No.11614311)
【分类号】：R965

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