微RNA-130a-3p对脂多糖诱导的肺泡上皮细胞炎性因子的保护作用及机制研究  

Protective effect of miR-130a-3p on LPS-induced alveolar epithelial cell inflammation and its mechanism

作　　者：曹军[1] 王丽霞[1] 范小勇[1] Cao Jun;Wang Lixia;Fan Xiaoyong(Department of Emergency,Xinxiang Central Hospital,Xinxiang,Henan 453000,China)

机构地区：[1]新乡市中心医院急诊科,河南453000

出　　处：《中华生物医学工程杂志》2019年第5期552-557,共6页Chinese Journal of Biomedical Engineering

摘　　要：目的探讨微RNA-130a-3p(miR-130a-3p)对脂多糖(LPS)诱导的肺泡上皮细胞A549炎性因子的保护作用及机制研究。方法不同浓度的LPS处理A549细胞,MTT检测细胞活性,实时荧光定量PCR(qRT-PCR)检测miR-130a-3p水平。LPS诱导miR-130a-3p过表达的A549细胞,MTT检测细胞活性,流式细胞仪检测细胞凋亡,免疫印迹法检测缺氧诱导因子1α基因(HIF1A)、IL-1β、IL-6、TNF-α蛋白水平。双荧光素酶报告基因实验验证miR-130a-3p与HIF1A关系。结果LPS处理后,A549细胞活性和miR-130a-3p表达水平显著降低(P<0.05),IL-1β、IL-6和TNF-α蛋白水平、A549细胞凋亡率显著升高(P<0.05)。过表达miR-130a-3p提高了LPS诱导的A549细胞活性,降低了LPS诱导的A549细胞IL-1β、IL-6和TNF-α蛋白水平和凋亡率(P<0.05)。miR-130a-3p负调控HIF1A表达,过表达HIF1A降低了过表达miR-130a-3p对LPS诱导的A549细胞活性、凋亡率及IL-1β、IL-6和TNF-α蛋白表达的影响。结论miR-130a-3p通过抑制HIF1A表达降低LPS诱导的肺泡上皮细胞A549凋亡和炎性反应。Objective To investigate the protective effect and its mechanism of miR-130a-3p on lipopolysaccharide(LPS)-induced alveolar epithelial A549 inflammation.Methods A549 cells were treated with LPS at different doses.Cell viability was detected by MTT,and miR-130a-3p level was detected by qRT-PCR.miR-130a-3p overexpressing A549 cells were established by LPS induction.Then,cell viability was detected by MTT,cell apoptosis by flow cytometry,and the levels of hypoxia-inducible factor 1α gene(HIF1A),IL-1β,IL-6 and TNF-α by Western Blotting.Dual luciferase reporter gene assay was used to verify the targeting relationship between miR-130a-3p and HIF1A.Results After LPS treatment,the viability of A549 cells and the expression of miR-130a-3p were significantly decreased(P<0.05),the levels of IL-1β,IL-6 and TNF-α,and the apoptosis rate of A549 cells were significantly increased(P<0.05).Overexpression of miR-130a-3p increased LPS-induced A549 cell viability,decreased the protein levels of IL-1β,IL-6 and TNF-α,and inhibited apoptosis in A549 cells(P<0.05).miR-130a-3p negatively regulated HIF1A expression.Overexpression of HIF1A attenuated the effect of miR-130a-3p overexpression on LPS-induced A549 cell viability,apoptosis and protein expession of IL-1β,IL-6 and TNF-α.Conclusion miR-130a-3p may attenuate LPS-induced apoptosis and inflammatory response of alveolar epithelial cells A549 by inhibiting HIF1A expression.

关 键 词：微RNA-130a-3p 缺氧诱导因子1α基因 脂多糖 肺泡上皮细胞 炎性因子 

分 类 号：R563[医药卫生—呼吸系统]