摘要
目的 明确内毒素脂多糖（LPS）对microRNA-142-3p（miR-142-3p）的表达调控作用及miR-142-3p的作用靶点.方法 体外培养人单核细胞株THP-1,用LPS刺激48 h后,real-time RT-PCR方法检测miR-142-3p和腺苷酸环化酶9（AC-9）的表达变化.接着采用脂质体转染的方法,将miR-142-3p的拟似物（mimic）和抑制剂（inhibitor）转染进入THP-1细胞,观察AC-9的表达变化.应用SPSS13.0统计软件,采取t检验进行数据统计.结果 THP-1细胞受LPS刺激48 h后,miR-142-3p的表达上调（P＜0.05）,AC-9表达则减少（P＜0.05）.上调miR-142-3p水平后,细胞内AC-9的表达水平出现下降（P＜0.05）,而抑制miR-142-3p的水平后,细胞内AC-9的表达水平上升（P＜0.05）.结论 miR-142-3p可调控THP-1细胞AC-9基因表达水平,且可能具有类似LPS的促炎机制,提示miR-142-3p在调控免疫细胞的炎症反应过程中具有重要的作用.
Objective To investigate the regulatory effect of lipopolysaccharides （LPS） with regard to the expression of miR-142-3p and identify the target of miR-142-3p in THP-1 cells.Methods In our study,human monocytes THP-1 cell line was stimulated for 48 hours with 0.5 μ g/ml of LPS,transfected with miR-142-3p mimic and inhibitor （100 nM） by lipofectamine RNAiMAX.The levels of miR-142-3p and AC-9 mRNA in the THP-1 cells were detected by real-time RT-PCR.Statistical analysis was carried out by using SPSS 13.0 software,t-test was used.Results The level of AC-9 mRNA decreased and the level of miR-142-3p increased compared with that of the control group after 0.5 μ g/ml LPS stimulated.After 100 nM miR-142-3p mimic transfection,AC-9 mRNA level in THP-1 cells reduced,and after miR-142-3p inhibitor transfection,AC-9 mRNA level increased.Conclusion miR-142-3p may regulate THP-1 cells＇inflammation response liked LPS by adjusting the AC-9 gene expression.miR-142-3p may play an important role in the inflammatory response of immune cells.
出处
《国际医药卫生导报》
2013年第20期3087-3091,共5页
International Medicine and Health Guidance News
基金
广东省自然科学基金项目（10151018201000018
S2012010008701）
