LncRNA MINCR靶向miR⁃223抑制LPS诱导的人肺泡上皮细胞系A549损伤和炎性反应  

LncRNA MINCR inhibits LPS-induced damage and inflammation of alveolar epithelial cell line A549 by targeting at miR-223

作　　者：杨广林 任丽娟 陈文静 熊维军 YANG Guanglin;REN Lijuan;CHEN Wenjing;XIONG Weijun(Department of Respiratory Medicine,Affiliated Hospital of Sichuan Nursing Vocational College/the Third People’s Hospital of Sichuan Province,Chengdu 610100,China)

机构地区：[1]四川护理职业学院附属医院/四川省第三人民医院呼吸内科,四川成都610100

出　　处：《基础医学与临床》2023年第1期102-109,共8页Basic and Clinical Medicine

基　　金：四川省卫生和计划生育委员会科研课题(18PJ108)。

摘　　要：目的探讨长链非编码RNA(lncRNA)MINCR靶向调节微小RNA(miR)-223对脂多糖(LPS)诱导的肺泡上皮细胞系损伤和炎性反应的影响。方法将人肺泡上皮细胞系A549随机分为对照组、LPS组、LPS+转染组(Si NC组、Si MINCR组、miR-223 NC组、miR-223 mimic组、Si NC+miR-223 NC组、Si NC+miR-223 antagomir组、Si MINCR+miR-223 NC组、Si MINCR+miR-223 antagomir组)。RT-qPCR检测MINCR、miR-223表达水平;流式细胞仪检测细胞凋亡率;ELISA测定血清中肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)水平;生物信息学预测和双荧光素酶报告基因验证MINCR与miR-223的靶向关系;Western blot检测细胞中活化的半胱天冬氨酸蛋白酶-3(cleaved caspase-3)、B细胞淋巴瘤-2(Bcl-2)蛋白表达水平。结果MINCR和miR-223存在靶向关系。LPS组与对照组比较,细胞凋亡率、TNF-α、IL-6水平、MINCR、cleaved caspase-3蛋白表达均显著增加(P<0.05),miR-223、Bcl-2蛋白表达显著降低(P<0.05)。与Si NC组相比,Si MINCR组细胞凋亡率、TNF-α和IL-6水平、MINCR和cleaved caspase-3蛋白表达均显著降低(P<0.05),Bcl-2蛋白表达显著升高(P<0.05);与miR-223 NC相比,miR-223 mimic组细胞凋亡率、TNF-α和IL-6水平、cleaved caspase-3蛋白表达均显著降低(P<0.05),miR-223、Bcl-2蛋白表达显著升高(P<0.05)。miR-223 antagomir可逆转Si MINCR发挥的作用(P<0.05)。结论沉默lncRNA MINCR的表达可能通过靶向激活miR-223表达,抑制A549细胞凋亡以及炎性反应,减少LPS诱导的A549细胞损伤。Objective To investigate the potential effects of long non-coding RNA(lncRNA)MINCR on lipopolysaccharide(LPS)-induced human alveolar epithelial cell line A549 damage and inflammation through targeted regulation of miR-223.Methods A549 cells were randomly divided into control group,LPS group,LPS+transfection group(Si NC group,Si MINCR group,miR-223 NC group,miR-223 mimic group,Si NC+miR-223 NC group,Si NC+miR-223 antagomir group,Si MINCR+miR-223 NC group,and Si MINCR+miR-223 antagomir group).RT-qPCR was performed to determine the expression levels of MINCR and miR-223;Flow cytometry was performed to determine the rate of apoptosis;ELISA was performed to determine the levels of serum tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6);Bioinformatics prediction and dual luciferase reporter gene assay were performed to verify the targeting relationship of MINCR and miR-223;Western blot was performed to test the expression levels of activated caspase-3(cleaved caspase-3)and B-cell lymphoma-2(Bcl-2)proteins in the cells.Results There was a targeting relationship between MINCR and miR-223.Compared with the control group,the apoptosis rate,the level of TNF-αand IL-6,and the expression of MINCR,cleaved caspase-3 proteins in the LPS group were significantly increased(P<0.05).The expression of miR-223 and Bcl-2 proteins significantly decreased(P<0.05).Compared with the Si NC group,the apoptosis rate,the levels of TNF-αand IL-6,and the expression of MINCR and cleaved caspase-3 proteins in the Si MINCR group were obviously reduced(P<0.05);And the expression of Bcl-2 protein was obviously increased(P<0.05).Compared with miR-223 NC,the apoptosis rate,the levels of TNF-αand IL-6,and the expression of cleaved caspase-3 protein in the miR-223 mimic group were obviously reduced(P<0.05);And the expression of miR-223 and Bcl-2 proteins was obviously increased(P<0.05).miR-223 antagomir was able to reverse the effect of Si MINCR(P<0.05).Conclusions Silencing the expression of lncRNA MINCR may target the activation of miR-223 exp

关 键 词：肺泡上皮细胞 lncRNA MINCR miR-223 炎性反应 细胞损伤 

分 类 号：R563[医药卫生—呼吸系统]