lncRNA PACER促进脓毒症急性肺损伤炎症反应的实验研究

Experimental study of lncRNA PACER promoting inflammatory response of acute lung injury induced by sepsis

目的探讨lncRNA PACER对小鼠脓毒症急性肺损伤炎症反应的促进作用。方法分离和培养急性肺损伤和健康非吸烟者的肺泡巨噬细胞,以及获取细菌脂多糖(LPS)诱导的急性肺损伤(ALI)小鼠肺组织,采用qRT-PCR方法检测lncRNA PACER的表达;过表达或敲低PACER后,ELISA方法检测THP-1和RAW264.7细胞炎性因子肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)的表达;对LPS所致ALI小鼠,尾静脉注射PACER siRNA慢病毒后,ELISA检测小鼠血清炎性因子TNF-α、IL-6的表达,HE染色观察肺组织病理学变化。结果 ALI患者肺泡巨噬细胞和ALI小鼠肺组织中lncRNA PACER表达均显著升高(P<0.01);细胞过表达PACER后,细胞炎性因子肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)的表达升高,而敲低PACER后,炎性因子TNF-α、IL-6的表达则降低(P<0.01); ALI小鼠敲低PACER后,小鼠肺组织和血清中炎性因子TNF-α、IL-6的表达均显著降低(P<0.01),肺组织损伤程度明显减弱。结论 lncRNA PACER可显著促进脓毒症急性肺损伤炎症反应,为明确lncRNA PACER作为ALI防治的靶标提供了依据。

Objective To investigate the effect of lncRNA PACER on inflammatory response of acute lung injury induced by sepsis in mice.Methods Human alveolar macrophages were collected from 20 cases of acute lung injury and health nonsmokers,and Lung tissues of ALI mice induced by LPS were obtained to detect the expression of lncRNA PACER by qRT-PCR.Then,lncRNA PACER was overexpressed or knocked down in THP-1 and RAW264.7 cells,the expressions of cytoinflammatory cytokines TNF-αand IL-6 were detected by ELISA.Furthermore,the ALI mice induced by LPS were injected PACER siRNA lentivirus into the tail vein,and the expressions of serum inflammatory factors TNF-αand IL-6 were detected by ELISA,meanwhile,the pathologic injury of lung tissue were observed by HE staining.Results The expression of lncRNA PACER was significantly increased in both alveolar macrophages of ALI patients and in lung tissues of ALI mice(P<0.01).After overexpression of PACER,the expression of inflammatory cytokines TNF-αand IL-6 were increased,while the expression of inflammatory cytokines TNF-αand IL-6 decreased after knocked down PACER(P<0.01).The expression of inflammatory cytokines TNF-αand IL-6 in lung tissue and serum of ALI mice were significantly decreased after the mice were knocked down PACER(P<0.01),and the pathological damage in lung tissues was significantly alleviated in PACER knockdown mice.Conclusions LncRNA PACER significantly promotes the inflammatory response of acute lung injury in sepsis,which provides basis for the identification of lncRNA PACER as a target for the prevention and treatment of ALI.