摘要
目的 探讨白细胞介素 1β (IL 1β)是否引起细胞内蛋白聚集体的形成以及细胞活性氧 (ROS)含量的变化和线粒体膜电位的改变。方法 用不同浓度的IL 1β处理神经胶质瘤U2 5 1细胞 2 4h ,四唑盐 (MTT)比色法测定线粒体氧化还原酶的活性 ,用硫磺素S染色研究细胞内蛋白聚集体形成情况 ,四氯四乙基苯并咪唑基羰花青碘化物 (JC 1)检测线粒体膜电位 ,二氯荧光黄双乙酸盐(DCFH DA)检测细胞内活性氧水平。结果 5ng/ml的IL 1β作用于U2 5 1小于 6h没有降低细胞活性。不同浓度IL 1β处理 2 4h后 ,与对照组比较 ,U2 5 1细胞活性在 5ng/ml时明显下降 (P <0 0 5 ) ,并随着IL 1β剂量继续加大 ,细胞存活率逐渐降低。IL 1β (5 0ng/ml)可导致细胞内蛋白聚集体形成增多 ,并且用JC 1荧光染色可检测到线粒体膜电位明显升高 ,用DCFH DA荧光染色和流式细胞仪定量检测显示 ,U2 5 1细胞内ROS含量增加。结论 IL 1β可引起U2 5 1细胞内蛋白聚集体形成 ,线粒体膜电位增高 ,ROS含量增加进而细胞氧化应激反应增强 。
Objective To study the effects of IL-1β on the formation of intracellular protein aggregates, mitochondria membrane potential and reactive oxygen species (ROS) levels in neuroglioma U251 cells. Methods Neuroglioma U251 was exposed to different concentrations of IL-1β for 24 hours in vitro. Mitochondria reductase viability was assessed with MTT. Thioflavine S staining was used to study the formation of intracellular protein aggregates. Mitochondria membrane potential and ROS levels were detected with 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethylbenzimidazolylcarbocyanine iodide (JC-1) and 2′,7′-dichlorofluorescin diacetate (DCFH-DA) respectively.Results IL-1β (5 ng/ml) did not decrease mitochondria reductase viability under 6 hours, but after treatment with IL-1β (5 ng/ml) for 24 hours, mitochondria reductase viability declined significantly (P<0.05) as compared with the controls. IL-1β (50 ng/ml) induced the formation of protein aggregates. Depolarization of mitochondria potential was found by using JC-1 fluorescence staining, and the increase in ROS levels was also shown with DCFH-DA fluorescence stain and flow cytometry (FCM) after treatment with IL-1β (50 ng/ml) on U251 cells. Conclusions In neuroglioma U251, IL-1β (50 ng/ml) could induce the formation of protein aggregates, resulting in intracellular ROS increase, mitochondria potential depolarization, oxidative stress and nerve cell death eventually.
出处
《中华神经科杂志》
CAS
CSCD
北大核心
2004年第5期396-401,共6页
Chinese Journal of Neurology
基金
国家重点基础研究规划资助项目 (G19990 5 40 0 8)
国家自然科学基金资助项目 ( 3 9970 2 63
3 0 1710 2 5 )
上海市卫生系统百名跨世纪优秀学科带头人培养计划资助项目 ( 97BR0 0 1)
上海市科委"启明星后"计划资助项目 (97QMB14 10 )
