miR-124通过p38α抑制小胶质细胞分泌促炎细胞因子  

Micro RNA-124 downregulates pro-inflammatory cytokines in BV-2 microglia cells by targeting p38αMAPK

作　　者：朱志远[1] 卢国辉[2] 叶勇义[1] 何骁征[1] 张世忠[1] 

机构地区：[1]国家临床重点专科教育部工程技术研究中心,广东省脑功能修复与再生重点实验室南方医科大学珠江医院神经外科,广州510282 [2]南昌大学第一附属医院神经外科,南昌330006

出　　处：《中华神经医学杂志》2016年第6期563-568,共6页Chinese Journal of Neuromedicine

基　　金：国家自然科学基金（81371397、81560220）江西省青年科学基金（20151BAB215014）

摘　　要：目的探讨miR-124对小胶质细胞分泌促炎细胞因子的调控作用及其机制。方法（1）用不同浓度梯度脂多糖（LPS）和不同时间刺激BV-2小胶质细胞活化，RT-qPCR检测BV．2细胞中miR-124的表达。（2）将细胞均分为4组：PBS组、LPS组、LPS＋ctrl-模拟物组（LPS刺激后转染含无义序列的ctrl-模拟物）；LPS＋miR-124模拟物组（LPS刺激后转染miR-124模拟物）。RT-qPCR和ELISA检测各组细胞肿瘤坏死因子α（TNF-α）和白介素1β（IL-1β）mRNA和蛋白水平的表达变化．Western blotting检测p38α、细胞外调节蛋白激酶（ERK）及其磷酸化水平的变化。（3）在上述分组基础上增设4组：LPS＋VX702组、LPS＋miR-124抑制剂组、LPS＋VX702＋miR-124模拟物组、LPS＋VX702＋miR-124抑制剂组，后2组将BV-2细胞经p38α特异性抑制VX702预处理后．转染miR-124模拟物和抑制剂，LPS刺激其活化。ELISA检测TNF-α和IL-1β表达的变化。结果（1）LPS刺激后BV-2细胞中miR-124表达明显下降，与未刺激细胞相比，差异有统计学意义（P〈0．05），且呈浓度和时间依赖性。（2）与LPS＋ctrl-模拟物组相比，LPS＋miR-124模拟物组细胞TNF-α和IL-1βmRNA、蛋白的表达水平均显著降低，p38α及p-p38α的表达量亦显著降低，差异均有统计学意义（P〈0．05）；但2组间ERK及p-ERK的表达差异元统计学意义（P〉0．05）。0）VX702预处理后，LPS＋vX702＋miR-124模拟物组细胞与LPS＋VX702组细胞相比，TNF-α和IL-1β的分泌量差异无统计学意义（P〉0．05）。LPS＋VX702＋miR-124抑制剂组与LPS＋VX702组相比，TNF-α和IL-1β的表达差异亦无统计学意义（P〉0．05）。结论过表达miR-124可抑制LPS诱导的促炎细胞因子分泌，其机制可能与调控p38α的表达有关。Objective To investigate the role of micro RNA-124 （miR-124） in regulating activation of microglias and secretion of pro-inflammation cytokines and its potential mechanism. Methods （1） BV-2 cells were exposed to different concentration of lipopolysaccharide （LPS） for different durations; relative expression level of miR-124 was detected by real time- quantitative PCR （RT-qPCR）. （2） The BV-2 cells were divided into four groups： PBS group, LPS group, LPS＋ctrl-simulant group and LPS＋miR-124 simulant group. Protein and mRNA expressions of inflammatory factors （tumor necrosis factor [TNF]-α and interleukin [IL] 1-β） were evaluated by RT-qPCR and ELISA. Expressions of p38a, phosphorylated （p）-p38α ERK and p-ERK were detected by Western blotting. （3） Besides the above groups, four groups were added： LPS＋VX702 group, LPS＋miR-124 inhibitor group, LPS＋VX702＋miR-124 simulant group and LPS＋VX702＋miR-124 inhibitor group; pretreatment with p38α specific inhibitor VX702 was given to the BV-2 cells, the latter two groups were given miR-124 simulant or miR-124 inhibitor, and LPS was used to activate the cells; the expressions of TNF-α and IL-1β were evaluated by ELISA. Results （1） As compared with control group, miR-124 was significantly down-regulated in LPS-stimulated BV-2 cells （P〈0.05）, in a dose- and time- dependent manner. （2） As compared with cells in the LPS＋ctrl-simulant group, cells in the LPS＋miR-124 simulant group had significantly decreased TNF-α and IL-1β mRNA and protein expressions, and p38α and p-p38α levels （P〈0.05）; the ERK and p-ERK levels showed no significant difference between the two groups （P〉0.05）. （3） The TNF-α and IL-1β levels between LPS＋VX702 group and LPS＋VX702＋miR-124 simulant group were not significantly different （P〉0.05）; those between the LPS＋VX702＋miR-124 inhibitor group and the LPS＋VX702 group were not significantly different （P〉0. 05）. Conclusion The miR-124 is down-regul

关 键 词：神经炎症 小胶质细胞 miR-124 p38α 

分 类 号：R741[医药卫生—神经病学与精神病学]