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

doi:10.16352/j.issn.1001-6325.2023.01.0102

基础医学与临床 ›› 2023, Vol. 43 ›› Issue (1): 102-109.doi: 10.16352/j.issn.1001-6325.2023.01.0102

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

杨广林, 任丽娟, 陈文静, 熊维军^*

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

收稿日期:2021-12-31 修回日期:2022-05-09 出版日期:2023-01-05 发布日期:2022-12-27
通讯作者: ^*763840401@qq.com
基金资助:
四川省卫生和计划生育委员会科研课题(18PJ108)

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

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

Received:2021-12-31 Revised:2022-05-09 Online:2023-01-05 Published:2022-12-27
Contact: ^*763840401@qq.com

摘要/Abstract

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。

关键词: 肺泡上皮细胞, lncRNA MINCR, miR-223, 炎性反应, 细胞损伤

Abstract: 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-α and 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-α and 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-α and 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 expression, inhibit cell apoptosis and inflammation, and reduce LPS-induced damage of alveolar epithelial cells.

Key words: alveolar epithelial cells, lncRNA MINCR, miR-223, inflammation, cell damage

中图分类号:

R563

杨广林, 任丽娟, 陈文静, 熊维军. 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[J]. 基础医学与临床, 2023, 43(1): 102-109.

YANG Guanglin, REN Lijuan, CHEN Wenjing, XIONG Weijun. LncRNA MINCR inhibits LPS-induced damage and inflammation of alveolar epithelial cell line A549 by targeting at miR-223[J]. Basic & Clinical Medicine, 2023, 43(1): 102-109.

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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

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