lncRNA LUCAT1靶向miR-203a-3p影响LPS诱导的Ⅱ型肺泡上皮细胞凋亡和炎症反应的分子机制研究  

Effects of lncRNA LUCAT1 targeting miR-203a-3p on LPS-induced apoptosis and inflammation of type II alveolar epithelial cells

作　　者：张佳婕[1] 李引钰[1] 秦进[2] ZHANG Jiajie;LI Yinyu;QIN Jin(Department of Clinical Laboratory,Affiliated Hospital of Chengdu University,Chengdu 610081,China;Department of Blood Transfusion,Affiliated Hospital of Chengdu University,Chengdu 610081,China)

机构地区：[1]成都大学附属医院检验科,610081 [2]成都大学附属医院输血科,610081

出　　处：《免疫学杂志》2023年第8期689-695,共7页Immunological Journal

摘　　要：目的探讨长链非编码RNA(lncRNA)LUCAT1靶向miR-203a-3p对LPS诱导的Ⅱ型肺泡上皮细胞凋亡和炎症反应的影响。方法qRT-PCR检测脓毒症急性肺损伤患者外周血中和不同浓度(7.5、15、30 mg/L)LPS处理Ⅱ型肺泡上皮细胞中lncRNA LUCAT1和miR-203a-3p表达水平。用30 mg/L LPS处理Ⅱ型肺泡上皮细胞,细胞分为NC组、LPS组、si-NC+LPS组、silncRNA LUCAT1+LPS组、miR-NC+LPS组、miR-203a-3p+LPS组、anti-miR-NC+si-lncRNA LUCAT1+LPS组、anti-miR-203a-3p+si-lncRNA LUCAT1+LPS组。q RT-PCR检测lncRNA LUCAT1和miR-203a-3p表达水平;流式细胞术检测细胞凋亡;Western blot检测Cleaved-caspase-3蛋白表达;ELISA检测TNF-α、IL-6、IL-1β含量;双荧光素酶报告实验检测lncRNA LUCAT1和miR-203a-3p靶向关系。结果在脓毒症急性肺损伤患者外周血中及不同浓度LPS处理Ⅱ型肺泡上皮细胞中lncRNA LUCAT1表达水平增加,miR-203a-3p表达水平降低。LPS增加Ⅱ型肺泡上皮细胞凋亡率、Cleaved-caspase-3蛋白表达水平,增多TNF-α、IL-6、IL-1β含量;干扰lncRNA LUCAT或过表达miR-203a-3p降低LPS诱导的Ⅱ型肺泡上皮细胞凋亡率、Cleaved-caspase-3蛋白表达水平,减少TNF-α、IL-6、IL-1β含量。lncRNA LUCAT1通过调控miR-203a-3p表达,抑制anti-miR-203a-3p可以逆转干扰lncRNA LUCAT1对LPS诱导的Ⅱ型肺泡上皮细胞凋亡和炎症反应的影响。结论lncRNA LUCAT1通过靶向负调控miR-203a-3p减弱LPS诱导的Ⅱ型肺泡上皮细胞凋亡和炎症反应。This paper mainly discusses the effect of long non-coding RNA(lncRNA)LUCAT1 targeting miR-203a-3p on LPS-induced apoptosis and inflammatory response of type II alveolar epithelial cells.qRT-PCR was used to detect the expression levels of lncRNA LUCAT1 and miR-203a-3p in LPS(7.5,15,30 mg/L)-treated type II alveolar epithelial cells and the peripheral blood of patients with sepsis and acute lung injury.TypeⅡalveolar epithelial cells treated with 30 mg/L LPS were divided into NC group,LPS group,si-NC+LPS group,si-lncRNA LUCAT1+LPS group,miR-NC+LPS group,miR-203a-3p+LPS group,anti-miR-NC+si-lncRNA LUCAT1+LPS group,anti-miR-203a-3p+si-lncRNA LUCAT1+LPS group.qRT-PCR was used to detect the expression levels of lncRNA LUCAT1 and miR-203a-3p;flow cytometry was used to detect cell apoptosis;Western blot was used to detect Cleaved-caspase-3 protein expression;ELISA was used to detect TNF-α,IL-6 and IL-1βcontent;the dual luciferase reporter experiment was used to detect the targeting relationship between lncRNA LUCAT1 and miR-203a-3p.Data showed that the expression level of lncRNA LUCAT1 was increased but the expression level of miR-203a-3p was decreased in LPS-treated type II alveolar epithelial cells and the peripheral blood of patients with sepsis and LPS.LPS could increase the apoptosis rate,the expression of Cleaved-caspase-3 protein and the content of TNF-α,IL-6 and IL-1βin type II alveolar epithelial cells,while lncRNA LUCAT interfering or miR-203a-3p overexpression could reduce the LPS-induced effects on type II alveolar epithelial cells.LncRNA LUCAT1,by regulating the expression of miR-203a-3p,inhibited antimiR-203a-3p,which can reverse the effect of lncRNA LUCAT1 interfering on LPS-induced type II alveolar epithelial cell apoptosis and inflammation.Taken together,lncRNA LUCAT1 can attenuate LPS-induced type II alveolar epithelial cell apoptosis and inflammation through targeting negative regulation of miR-203a-3p.

关 键 词：LUCAT1 miR-203a-3p LPS Ⅱ型肺泡上皮细胞 凋亡 炎症反应 

分 类 号：R563.1[医药卫生—呼吸系统]