GPRC5A对LPS诱导的支气管上皮16HBE细胞损伤的影响及机制  

Effects and Mechanism of GPRC5A on LPS-Induced Bronchial Epithelial 16HBE Cell Injury

作　　者：王静[1] 康媛洁 刘翠翠[1] 石晓岚[1] 马彩玲[1] WANG Jing;KANG Yuanjie;LIU Cuicui;SHI Xiaolan;MA Cailing(Respiratory Asthma Center,Xi an Children s Hospital,Xi an 710003,China;Second Department of Respiratory Medicine,Xi an Children s Hospital,Xi an 710003,China)

机构地区：[1]西安市儿童医院呼吸哮喘中心,西安710003 [2]西安市儿童医院呼吸二科,西安710003

出　　处：《福建医科大学学报》2022年第4期317-323,共7页Journal of Fujian Medical University

摘　　要：目的探究G蛋白偶联受体家族C组5型(GPRC5A)对脂多糖(LPS)诱导的人支气管上皮16HBE细胞多种生物学特性的影响,并探讨其分子机制。方法利用LPS建立16HBE细胞损伤模型,将细胞分为4组:Control组、LPS组、LPS+pcDNA组和LPS+pcDNA-GPRC5A组。利用患者样本和基因表达组学数据库数据分析支气管哮喘(BA)患者和健康受试者气道组织中的GPRC5A的表达情况;采用实时荧光定量聚合酶链反应法检测GPRC5A、肿瘤坏死因子α(TNF-α)和白细胞介素6(IL-6)的mRNA表达水平;采用MTT法和流式细胞仪检测细胞增殖与凋亡;Western-Blot检测GPRC5A、气道黏蛋白5AC(MUC5AC)、NF-κB p65和NF-κB抑制因子α(IκBα)蛋白的表达水平。结果成功构建LPS诱导的16HBE细胞损伤模型;与健康受试者组织和Control组细胞比较,GPRC5A在EBCs组织和LPS组的16HBE细胞中下调(P<0.05);而与LPS组和LPS+pcDNA组比较,LPS+pcDNA-GPRC5A组的细胞凋亡率、炎症因子TNF-α和IL-6、气道黏蛋白MUC5AC、NF-κB p65和IκBα的表达均显著下降,而细胞活力升高(P<0.05)。结论GPRC5A可能通过NF-κB通路抑制LPS诱导的16HBE细胞凋亡、炎症反应和气道黏蛋白的分泌,促进细胞的活力,发挥阻碍损伤的作用。Objective To investigate the effect of G protein coupled receptor family C group 5 member A(GPRC5A)on lipopolysaccharide(LPS)-induced bronchial epithelial 16HBE cell injury and its mechanism.Methods The 16HBE cell injury model was established by LPS stimulation.The 16HBE cells were divided into:Control group,LPS group,LPS+pcDNA group,LPS+pcDNA-GPRC5A group.The expression of GPRC5A in the airway tissues of asthma patients(n=28)and normal subjects(n=42)was analyzed using the GEO dataset;The mRNA expression levels of GPRC5A,tumor necrosis factorα(TNF-α),and interleukin-6(IL-6)in 16HBE cells were measured by qRT-PCR;The cell viability of 16HBE cells was detected using MTT assay.The apoptosis of 16HBE cells was detected by flow cytometry.The protein expression levels of GPRC5A,airway mucin 5 subtype AC(MUC5AC),NF-κB p65 and inhibitory kappa B alpha(IκBα)in 16HBE cells were measured using Western-blot.Results The 16HBE cell injury model was successfully established by LPS stimulation.Compared with control group and normal tissues,the mRNA and protein expression levels of GPRC5A were significantly decreased in asthma tissues and LPS-induced 16HBE cells(P<0.05).After GPRC5A overexpression,LPS-induced apoptosis and the expression levels of TNF-α,IL-6,MUC5AC,NF-κB p65 and IκBαwere significantly reduced,while LPS-decreased cell viability was significantly enhanced in 16HBE cells(P<0.05).Conclusion GPRC5A alleviated 16HBE cell injury through NF-κB pathway by suppressing LPS-induced apoptosis,inflammation and mucin MUC5AC expression.

关 键 词：支气管上皮细胞 脂多糖 G蛋白偶联受体家族C组5型 炎症 细胞凋亡 

分 类 号：R562.25[医药卫生—呼吸系统]