circVMA21靶向miR-497-5p/MUC1轴对LPS诱导的肺上皮细胞炎症因子表达和细胞凋亡的影响  

Effect of circVMA 21 targeting miR-497-5 p/MUC 1 axis on expressions of inflammatory factors and apoptosis of lung epithelial cells induced by LPS

作　　者：刘国元 刘宇花 杨旭芳 LIU Guoyuan;LIU Yuhua;YANG Xufang(Department of Pediatrics,Xinzhou Second People's Hospital,Xinzhou 034100,China;Department of Pharmacy,Xinzhou Second People's Hospital,Xinzhou 034100,China;Department of Pediatrics,Xinzhou Traditional Chinese Medicine Hospital,Xinzhou 034099,China)

机构地区：[1]忻州市第二人民医院儿科,忻州034100 [2]忻州市第二人民医院药剂科,忻州034100 [3]忻州市中医院儿科,忻州034099

出　　处：《中国免疫学杂志》2025年第2期342-350,共9页Chinese Journal of Immunology

摘　　要：目的:探究环状RNA VMA21(circVMA21)是否靶向miR-497-5p/黏蛋白(MUC1)影响LPS诱导的肺上皮细胞炎症因子表达和细胞凋亡。方法:肺上皮细胞A549分为Con组(对照)、LPS组(LPS损伤)、LPS+pcDNA组、LPS+pcDNA-circ VMA21组、LPS+anti-miR-NC组、LPS+anti-miR-497-5p组、LPS+pcDNA-circVMA21+miR-NC组、LPS+pcDNA-circVMA21+miR-497-5p组。qRT-PCR测定circVMA21和miR-497-5p表达,ELISA测定炎症因子IL-6、IL-1β和TNF-α表达,流式细胞术测定细胞凋亡,Western blot测定MUC1、TLR4/NF-κB通路和凋亡相关cleaved-caspase3、cleaved-caspase9蛋白表达。双荧光素酶报告实验测定circVMA21与miR-497-5p、miR-497-5p与MUC1的靶向关系。结果:与Con组比较,LPS组肺上皮细胞中circVMA21、MUC1表达量减少,miR-497-5p表达量、IL-6、IL-1β、TNF-α表达水平、细胞凋亡率和凋亡蛋白cleaved-caspase3、cleaved-caspase9及TLR4、p-NF-κB p65/NF-κB p65表达增加(P<0.05)。与LPS+pcDNA组比较,LPS+pcDNA-circVMA21组肺上皮细胞中circVMA21、MUC1表达升高,IL-6、IL-1β、TNF-α表达水平、细胞凋亡率和蛋白cleaved-caspase3、cleaved-caspase9及TLR4、p-NF-κB p65/NF-κB p65表达降低(P<0.05)。circVMA21靶向调控miR-497-5p表达。与LPS+anti-miR-NC组比较,LPS+anti-miR-497-5p组肺上皮细胞中MUC1表达升高,IL-6、IL-1β、TNF-α表达水平、细胞凋亡率和蛋白cleaved-caspase3、cleaved-caspase9及TLR4、p-NF-κB p65/NF-κB p65表达量降低(P<0.05)。miR-497-5p靶向调控MUC1表达。与LPS+pcDNA-circVMA21+miR-NC组比较,LPS+pcDNA-circVMA21+miR-497-5p组肺上皮细胞中MUC1表达降低,IL-6、IL-1β、TNF-α表达水平、细胞凋亡率和蛋白cleaved-caspase3、cleaved-caspase9及TLR4、p-NF-κB p65/NF-κB p65表达升高(P<0.05)。结论:circVMA21可能通过靶向下调miR-497-5p促进MUC1表达,抑制TLR-4/NF-κB通路活化,进而减少LPS诱导的肺上皮细胞炎症因子表达和细胞凋亡。Objective:To investigate whether circular RNA VMA21(circVMA21)targeting miR-497-5p/mucin1(MUC1)affecting the expressions of inflammatory factors and apoptosis in lung epithelial cells induced by LPS.Methods:A549 lung epithelial cells were divided into Con group(control),LPS group(LPS injury),LPS+pcDNA group,LPS+pcDNA-circVMA21 group,LPS+anti-miR-NC group,LPS+anti-miR-497-5p group,LPS+pcDNA-circVMA21+miR-NC group,LPS+pcDNA-circVMA21+miR-497-5p group.Expressions of circVMA21 and miR-497-5p were measured by qRT-PCR,expressions of inflammatory factors IL-6,IL-1βand TNF-αwere determined by ELISA,flow cytometry was used to monitor cell apoptosis,and Western blot was employed to assay expressions of MUC1,TLR4/NF-κB pathway and apoptosis-related activation-cleaved-caspase3 and cleaved-caspase9 protein.The targeting relationships between circVMA21 and miR-497-5p,miR-497-5p and MUC1 were determined by dual luciferase reporter assay.Results:Compared with Con group,expressions of circVMA21 and MUC1 in lung epithelial cells of LPS group were decreased,while expression levels of miR-497-5p,IL-6,IL-1βand TNF-α,apoptosis rate,expressions of apoptotic proteins cleaved-caspase3 and cleaved-caspase9,and TLR4,p-NF-κB p65/NF-κB p65 were increased(P<0.05).Compared with LPS+pcDNA group,expressions of circVMA21 and MUC1 in lung epithelial cells of LPS+pcDNA-circVMA21 group were increased,while expression levels of IL-6,IL-1βand TNF-α,apoptosis rate,expressions of cleaved-caspase3 and cleaved-caspase9,and TLR4,p-NF-κB p65/NF-κB p65 were decreased(P<0.05).circVMA21 was targeted regulating the expression of miR-497-5p.Compared with LPS+anti-miR-NC group,expressions of MUC1 in lung epithelial cells of LPS+anti-miR-497-5p group was increased,while expression levels of IL-6,IL-1β,TNF-α,apoptosis rate,protein cleaved-caspase3 and cleaved-caspase9,and TLR4,p-NF-κB p65/NF-κB p65 were decreased(P<0.05).miR-497-5p was targeted regulating the expression of MUC1.Compared with LPS+pcDNA-circVMA21+miR-NC group,expression of MUC1 in lung

关 键 词：肺上皮细胞 circVMA21 miR-497-5p 黏蛋白1 炎症 细胞凋亡 

分 类 号：R563.8[医药卫生—呼吸系统]