MicroRNA-146a在脂多糖诱导的肺泡巨噬细胞中表达的动态变化

The expression changes of miR-146a in lipopolysaccharide-induced alveolar macrophages

目的 观察并分析microRNA-146a（miR-146a）在脂多糖（LPS）刺激肺泡巨噬细胞中不同时间表达的变化,为探讨miR-146a对肺泡巨噬细胞炎症反应的调控作用及机制奠定基础.方法 体外培养的肺泡巨噬细胞系NR8383分成LPS刺激组和磷酸盐缓冲液（PBS）对照组,1 μg/mL的LPS刺激3 h,6 h,12 h后采用酶联免疫吸附试验（ELISA）检测细胞上清液中TNF-α的表达水平,实时荧光定量PCR（Taqman探针法）检测细胞miR-146a的表达情况.运用SPSS13.0统计软件,采取单因素方差分析或t检验进行数据统计分析.结果 ①LPS刺激3 h,6 h,12 h,细胞上清液中TNF-α的表达比对照组明显升高（P＜0.01）;②LPS刺激6 h,12 h细胞中miR-146a表达增高（P＜0.01）,且呈持续增高趋势.结论 LPS刺激后,miR-146a在NR8383肺泡巨噬细胞中的表达上调,且随着刺激时间的延长,miR-146a的表达有增高的趋势,推测其可能参与了肺泡巨噬细胞的炎症反应调控.

Objective To explore the mechanism and effect of miR-146a on alveolar macrophages and to observe the changes of miR-146a expression in the LPS-induced alveolar macrophages. Method NR8383 alveolar macrophages were divided into LPS-stimulated group and control group, and the cells of former group were treated with LPS （ 1 μg/mL） and then incubated for 3 h, 6 h and 12 h, respectively. The level of TNF-α in the supernatant of cells was assayed by using enzyme-linked immunosorbent assay （ELISA）, and the expression of miR-146a of cells was detected by using Real-Time PCR （TaqMan probe）.Statistical analysis carried out by using SPSS 13.0 software package in which One-way ANOVA and Student＇s t-test were used. Results Compared with control group, the levels of TNF-α in the supernatant of cells were significantly increased 3 h, 6 h and 12 h after LPS challenge （P 〈 0.01 ）. The expression of miR-146a increased 6 h and 12 h after LPS stimulation in NR8383 cells（ P 〈0.01 ）, and it had an upward tendency.Conclusions The expression of miR-146a in alveolar macrophages increases after LPS-stimulation. It hints miR-146a may be involved in the regulation of the inflammatory responses produced by alveolar macrophages.