lncRNA KCNQ1OT1缓解LPS诱导的肺炎症损伤

The effect of lncRNA KCNQ1OT1 on regulating the inflammatory response of pneumonia injury induced by LPS

为探讨长链非编码RNA(long non-coding RNA,lncRNA)KCNQ1对链/反义转录本1(KCNQ1 opposite strand/antisense transcript 1,KCNQ1OT1)在脓毒症急性肺损伤(acute lung injury,ALI)炎症反应中的作用及其可能机制,腹腔注射LPS(10 mg/kg)建立脓毒症ALI大鼠模型,体外建立LPS诱导的人小气道上皮细胞(human small airway epithelial cell,HSAEC)。采用ELISA检测肺组织中TNF-α、IL-6和IL-1β的相对水平,qRT-PCR和Western blotting检测KCNQ1OT1、miR-192-5p和凋亡相关蛋白(Caspase-3、Bax)的表达水平,CCK-8试验检测细胞增殖情况,生物信息学方法、双荧光素酶报告基因试验分别预测和验证KCNQ1OT1与miR-192-5p的相互作用。结果显示,脓毒症模型HSAEC中KCNQ1OT1显著下调,miR-192-5p显著上调(P<0.05)。KCNQ1OT1直接靶向miR-192-5p并负调控后者的表达。过表达KCNQ1OT1促进HSAEC的增殖,抑制细胞凋亡,而敲低KCNQ1OT1则发挥相反的作用(P<0.05)。该研究提示,KCNQ1OT1可通过抑制miR-192-5p表达减轻脓毒症ALI的炎症反应。

This study aims to investigate the role of long non-coding RNA(lncRNA)KCNQ1 opposite strand/antisense transcript 1(KCNQ1 OT1)in the inflammatory response in sepsis-induced acute lung injury(ALI)and its possible mechanism.Mouse models of ALI were established by intraperitoneal injection of LPS(10 mg/kg)and LPS-induced human small airway epithelial cells(HSAEC)were established in vitro.The relative levels of TNF-α,IL-6,and IL-1βin lung tissue were detected by ELISA.qRT-PCR and Western blotting were used to detect the expression levels of KCNQ1 OT1,miR-192-5 p,apoptosis-related proteins Caspase-3 and Bax.And CCK-8 assay was used to detect cell proliferation.Bioinformatics prediction and dual-luciferase report method predicted and verified the interaction between KCNQ1 OT1 and miR-192-5 p respectively.The results showed that KCNQ1 OT1 was significantly down-regulated in the sepsis model HSAECs,and miR-192-5 p was significantly up-regulated(P<0.05).KCNQ1 OT1 directly targeted miR-192-5 p and negatively regulated its expression.Overexpression of KCNQ1 OT1 promoted the proliferation of HSAECs and inhibited apoptosis while knocking down KCNQ1 OT1 had the opposite effect(P<0.05).In conclusion,KCNQ1 OT1 attenuates sepsis-mediated ALI by inhibiting expression of miR-192-5 p.