發(fā)布時(shí)間：2019-06-27 12:23

【摘要】：目的通過分析microRNA-146a在脂多糖(LPS)誘導(dǎo)慢性阻塞性肺疾病(COPD)大鼠遠(yuǎn)端肺動脈平滑肌細(xì)胞(PASMCs)中不同時(shí)間點(diǎn)表達(dá)的變化及與其合成分泌白細(xì)胞介素-6(IL-6)的相關(guān)性分析,探討microRNA-146a對PASMCs炎癥反應(yīng)的調(diào)控作用。方法建立經(jīng)典的COPD大鼠模型,采用酶消化聯(lián)合組織塊貼壁法,培養(yǎng)原代大鼠遠(yuǎn)端PASMCs,將第4代PASMCs分成對照組及LPS組,對照組不加入任何干預(yù)劑,LPS組用終濃度為1μg/m L的LPS誘導(dǎo)細(xì)胞;兩組細(xì)胞分別培養(yǎng)12、24、48、72 h后采用酶聯(lián)免疫吸附試驗(yàn)(ELISA)檢測細(xì)胞培養(yǎng)上清液中IL-6的表達(dá)水平;用實(shí)時(shí)熒光定量PCR(Taqman探針法)檢測細(xì)胞microRNA-146a的表達(dá)情況,并將細(xì)胞上清液中IL-6的表達(dá)量與microRNA-146a表達(dá)情況進(jìn)行相關(guān)性分析。結(jié)果與對照組相比,LPS誘導(dǎo)組PASMCs中microRNA-146a的表達(dá)量在12 h開始升高,并呈遞增趨勢,72 h達(dá)峰(P0.01);同時(shí),LPS組細(xì)胞上清液中IL-6的表達(dá)量亦于12 h時(shí)升高,并且升高明顯,72 h可達(dá)到高峰(P0.01);經(jīng)相關(guān)性分析,兩者的表達(dá)量呈正相關(guān)(r=0.981,P=0.00)。結(jié)論 COPD大鼠PASMCs在LPS誘導(dǎo)后合成分泌IL-6,同時(shí)PASMCs中microRNA-146a表達(dá)升高,兩者表達(dá)量呈正相關(guān),推測其可能與PASMCs的炎癥反應(yīng)調(diào)控有關(guān)。
[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，

本文編號：2506783