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  

The expression of microRNA-146a and the correlation analysis about synthesis and secretion of IL-6 in lipopolysaccharide-induced distal pulmonary artery smooth muscle cells of rats with COPD

作　　者：杨丹[1] 王昌明[1] 马礼兵[1] 林武洲[1] 徐青[1] 陈峰[1] 刘晓黎[1] 吕倩[1] 蒋明[1] 

机构地区：[1]桂林医学院附属医院呼吸内科,广西桂林541001

出　　处：《广东医学》2017年第23期3554-3557,共4页Guangdong Medical Journal

基　　金：国家自然科学基金资助项目(编号:81360010);广西医疗卫生重点科研课题(编号:重2012004)

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(P<0.01);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(P<0.01);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。Objective By analyzing the change of microRNA - 146a （miR - 146a） expression in lipopolysaccha- ride （LPS） - stimulated distal pulmonary artery smooth muscle cells （PASMCs） isolated from rats with chronic obstructive pulmonary disease （COPD） in different times and to investigate the role and mechanism of microRNA -146a on the in- flammatory response in the PASMCs. Methods The model of COPD was set up with uniting the enzyme digestion and tissue adherent method. The fourth generation of PASMCs were divided into control group and LPS groups. In LPS groups, the cells were treated with LPS （final concentration of 1 p.g/mL） and incubated for 12 hours （12 h）, 24 hours （24 h）, 48 hours （48 h） and 72 hours （72 h）, respectively. The interleukin -6 （IL-6） in the supernatant was assessed by u- sing enzyme - linked immunosorbent assay （ELISA）. And real - time PCR （ Taqman probe method） was used to assess microRNA - 146a expression. Statistical analysis was carried out by Student＇s t - test using SPSS18.0 software package. Results Compared with the control group, the expression of microRNA - 146a in PASMCs was increased after LPS in- duced in time - dependent manner. The expression of microRNA - 146a began to rise when induced for 12 hours, and reached the peak at the 72 hours （ P 〈 0. 01 ）. And so was the expression of IL - 6 in the supernatant （P 〈 0. 01 ）. Signif- icantly positive correlation between them was also revealed （P = 0. 00）. Conclusion The IL - 6 is increased with LPS stimulated in COPD distal PAMSCs, and so as the microRNA - 146a. They have a positive correlation, suggesting mi- croRNA - 146a is involved in regulating the inflammatory response of PAMSCs.

关 键 词：肺动脉平滑肌细胞 microRNA-146a 白细胞介素-6 脂多糖 炎症反应 

分 类 号：R543.2[医药卫生—心血管疾病]