摘要
目的探讨miR-129和miR-142—3p对人单核细胞株THP-1细胞自噬和凋亡的作用。方法体外培养人单核细胞株THP-1，化学合成人miR-129、miR-142—3p、miR-218、miR-214的拟似物（mimic），用脂质体LipofectamineRNAiMAX转染mimics进入THP-1细胞，同时用内毒素脂多糖（LPS，2μg／mL）刺激细胞，48h后收集细胞标本；采用流式细胞仪方法检测细胞凋亡及存活率，westernblotting方法检测自噬相关蛋白p62的表达水平。结果THP-1细胞内过表达miR-129或miR-142—3p后，与对照组比较，THP-1细胞存活率下降，细胞凋亡率和坏死率升高（P〈0．05），白噬相关蛋白p62表达量（P〈0．05）；LPS刺激细胞后亦出现类似的效应。结论miR-129和miR-142—3p能调控THP-1细胞的凋亡和自噬水平，进而影响细胞存活，提示miR-129和miR-142—3p在调控免疫细胞存活过程中具有重要作用。
Objective To investigate the roles of miR - 129 and miR - 142 - 3p on autophagy and apoptosis in THP - 1 cells. Methods In our study, human monocytes THP - 1 cell line were stimulated with 2 μg/mL lipopolysaceharides （ LPS ） for 48 hours. Cells were also transfected with miRNAs mimics, including miR- 129, miR- 142-3p, miR-218 and miR-214 （100 nmol/L）, by lipofectamine RNAiMAX for 48 hours. THP - 1 cells apoptosis rate and survival rate were detected by flow eytometry. And the autophagy related p62 protein was detected by western blotting. Statistical analysis was carried out by using SPSS 15.0 software, and data were analyzed by Student＇s t - test or one- way ANOVA. Results Cells＂ survival rates were decreased, apoptosis rates were increased and the expression level of p62 protein was descended after over - expressing of miR - 129 and miR - 142 - 3p in THP- 1 cells compared with the control group （P 〈0.05）, and similar results were shown in LIPS induced THP - 1 cells. Conclusion Over - expression of miR - 129 and miR - 142 - 3p can regulate the cells autophagy and apoptosis in THP - 1 cells. These results suggest that miR - 129 and miR - 142 -3p have important roles in the regulation of immune cells survival.
出处
《中国急救医学》
CAS
CSCD
北大核心
2014年第1期62-66,共5页
Chinese Journal of Critical Care Medicine
基金
广东省自然科学基金项目（S2012010008971,S2012010008701）
广东省产业技术研究与开发资金计划项目（20128031800233）
