脂多糖干预时间与小胶质细胞生成IL-1β和TNF-α时效关系的研究  

Study on time-effect relationship of IL- 1β and TNF- α coming from microglia treated with LPS

作　　者：白玉娟[1] 李琦军[1] 周红艳[1] 李炎[1] 张国亮[1] 

机构地区：[1]河北省石家庄市第三医院,河北石家庄050011

出　　处：《现代中西医结合杂志》2016年第8期828-831,847,共5页Modern Journal of Integrated Traditional Chinese and Western Medicine

摘　　要：目的以原代培养的大鼠皮质小胶质细胞为研究对象,观察脂多糖(LPS)处理小胶质细胞时间与小胶质细胞生成白细胞介素-1β(IL-1β)和肿瘤坏死因子α(TNF-α)的时效关系,寻找能够使小胶质细胞产生大量IL-1β、TNF-α的LPS最佳干预时间,为建立最佳小胶质细胞炎症模型提供依据。方法培养原代大鼠皮质小胶质细胞,随机分为10组,5组为LPS组,以含LPS(终浓度为100 ng/m L)的无血清胶质细胞培养液分别孵育1 h、3 h、6h、12 h、24 h;每个LPS组均设平行对照,对照组用无血清胶质细胞培养液孵育,孵育时间同LPS组。孵育结束后,ELISA方法检测各组培养液中IL-1β和TNF-α蛋白含量,RT-PCR方法检测小胶质细胞中IL-1βmRNA和TNF-αmRNA表达水平。结果孵育原代小胶质细胞1 h、3 h、6 h、12 h、24 h后,LPS组培养液中IL-1β和TNF-α蛋白含量及小胶质细胞中IL-1βmRNA和TNF-αmRNA表达水平均明显高于对照组(P均<0.05);LPS组培养液中IL-1β和TNF-α蛋白含量及小胶质细胞中IL-1βmRNA和TNF-αmRNA表达水平随着处理时间延长逐渐上升,IL-1β在3 h达到高峰,TNF-α在6h时达到高峰,二者在6 h均处于较高水平,以后随时间延长逐渐下降,相邻LPS处理组间比较差异均有统计学意义(P均<0.05)。结论 LPS能够刺激小胶质细胞大量产生IL-1β和TNF-α,这种效应与LPS刺激时间相关,在刺激6 h时IL-1β和TNF-α均处于较高水平,LPS刺激6 h时是理想的小胶质细胞炎症模型。Objective Primary cerebral cortical microglia of rat was cultured and treated with LPS to observe the timeeffect relationship of IL- 1β and TNF- α with treatment time of LPS,in order to find when microglia generate a large amount of IL- 1β and TNF- α with the treatment to provide evidences for the establish of best microglia inflammatory models. Methods Primary cerebral cortical microglia of SD rat was cultured and divided into 10 groups randomly,five groups were treated with serum-free medium included LPS（ final concentration 100 ng / m L） for different time（ 1 h,3 h,6 h,12 h,24 h）. Parallel control groups were arranged to each above groups and treated with serum-free medium for the same time. And then,the protein levels of IL- 1β and TNF- α in the culture media were detected through ELISA and the expressions of IL- 1β mRNA and TNF- α mRNA in the microglia cells were detected through RT- PCR. Results After 1 h,3 h,6 h,12 h,24 h of treatment by LPS,the protein levels of IL- 1β and TNF- α in the culture media and mRNA expression levels of IL- 1β and TNF-α in the microglia cells of LPS groups increased remarkably compared with the control groups（ P〈 0. 05）; the time point of the highest expression of IL- 1β was 3 h,the time point of the highest expression of TNF- α was 6 h,both of their amount stayed at higher status at 6 h,and then declined little by little with the extension of the time,and the differences were significant between every groups（ P〈 0. 05）. Conclusion LPS can stimulate microglia cells to generate a large amount of IL- 1βand TNF- α and this is time-depended,both of their amount stay at higher status at 6 h. It is better model of inflamation when microglia was treated with LPS for 6 h.

关 键 词：脂多糖 小神经胶质细胞 白细胞介素-1Β 肿瘤坏死因子Α 

分 类 号：R33[医药卫生—人体生理学]