LPS对COPD大鼠肺动脉平滑肌细胞microRNA-146a的表达及合成分泌IL-6的诱导作用
发布时间：2019-06-27 12:23
【摘要】：目的通过分析microRNA-146a在脂多糖(LPS)诱导慢性阻塞性肺疾病(COPD)大鼠远端肺动脉平滑肌细胞(PASMCs)中不同时间点表达的变化及与其合成分泌白细胞介素-6(IL-6)的相关性分析,探讨microRNA-146a对PASMCs炎症反应的调控作用。方法建立经典的COPD大鼠模型,采用酶消化联合组织块贴壁法,培养原代大鼠远端PASMCs,将第4代PASMCs分成对照组及LPS组,对照组不加入任何干预剂,LPS组用终浓度为1μg/m L的LPS诱导细胞;两组细胞分别培养12、24、48、72 h后采用酶联免疫吸附试验(ELISA)检测细胞培养上清液中IL-6的表达水平;用实时荧光定量PCR(Taqman探针法)检测细胞microRNA-146a的表达情况,并将细胞上清液中IL-6的表达量与microRNA-146a表达情况进行相关性分析。结果与对照组相比,LPS诱导组PASMCs中microRNA-146a的表达量在12 h开始升高,并呈递增趋势,72 h达峰(P0.01);同时,LPS组细胞上清液中IL-6的表达量亦于12 h时升高,并且升高明显,72 h可达到高峰(P0.01);经相关性分析,两者的表达量呈正相关(r=0.981,P=0.00)。结论 COPD大鼠PASMCs在LPS诱导后合成分泌IL-6,同时PASMCs中microRNA-146a表达升高,两者表达量呈正相关,推测其可能与PASMCs的炎症反应调控有关。
[Abstract]:Objective to analyze the expression of microRNA-146a in (PASMCs) of distal pulmonary artery smooth muscle cells of rats with chronic obstructive pulmonary disease (COPD) induced by lipopolysaccharide (LPS) at different time points and its correlation with the synthesis and secretion of IL-6 (IL-6), and to explore the regulatory effect of microRNA-146a on PASMCs inflammation. Methods the classical COPD rat model was established. The fourth generation PASMCs was divided into control group and LPS group by enzyme digestion combined with tissue block adherent method. The fourth generation PASMCs was divided into control group and LPS group without any intervention. The cells in LPS group were induced by LPS at the final concentration of 1 渭 g / mL, and the cells in the two groups were cultured for 12, 24, 48, 72 hours later, the expression of IL-6 in the culture medium was detected by enzyme-linked immunosorbent assay (ELISA). The expression of microRNA-146a in cell culture was detected by real-time fluorescence quantitative PCR (Taqman probe, and the correlation between the expression of IL-6 in cell culture and the expression of microRNA-146a was analyzed. Results compared with the control group, the expression of microRNA-146a in PASMCs of LPS induced group began to increase at 12 h, and reached the peak at 72 h (P 0.01). At the same time, the expression of IL-6 in the culture medium of LPS group also increased at 12 h, and reached the peak at 72 h (P < 0.01), and there was a positive correlation between them (r 鈮，
本文编号：2506783
[Abstract]:Objective to analyze the expression of microRNA-146a in (PASMCs) of distal pulmonary artery smooth muscle cells of rats with chronic obstructive pulmonary disease (COPD) induced by lipopolysaccharide (LPS) at different time points and its correlation with the synthesis and secretion of IL-6 (IL-6), and to explore the regulatory effect of microRNA-146a on PASMCs inflammation. Methods the classical COPD rat model was established. The fourth generation PASMCs was divided into control group and LPS group by enzyme digestion combined with tissue block adherent method. The fourth generation PASMCs was divided into control group and LPS group without any intervention. The cells in LPS group were induced by LPS at the final concentration of 1 渭 g / mL, and the cells in the two groups were cultured for 12, 24, 48, 72 hours later, the expression of IL-6 in the culture medium was detected by enzyme-linked immunosorbent assay (ELISA). The expression of microRNA-146a in cell culture was detected by real-time fluorescence quantitative PCR (Taqman probe, and the correlation between the expression of IL-6 in cell culture and the expression of microRNA-146a was analyzed. Results compared with the control group, the expression of microRNA-146a in PASMCs of LPS induced group began to increase at 12 h, and reached the peak at 72 h (P 0.01). At the same time, the expression of IL-6 in the culture medium of LPS group also increased at 12 h, and reached the peak at 72 h (P < 0.01), and there was a positive correlation between them (r 鈮，
本文编号：2506783
本文链接：https://www.wllwen.com/yixuelunwen/huxijib/2506783.html
最近更新
教材专著
