上调miR-338-3p减轻白介素-13诱导的人支气管细胞系BEAS-2B损伤  

Up-regulation of miR-338-3p alleviates IL-13-induced injury of human bronchial cell line BEAS-2B

作　　者：付海卫[1] 郭微微 盛芬[1] 刘东红[1] FU Haiwei;GUO Weiwei;SHENG Fen;LIU Donghong(Department of Respiratory Medicine,Taizhou First People's Hospital,Taizhou 318020,China)

机构地区：[1]台州市第一人民医院呼吸内科,浙江台州318020

出　　处：《基础医学与临床》2025年第3期346-353,共8页Basic and Clinical Medicine

基　　金：2023年台州市第二批社会发展科技计划(23ywb49)。

摘　　要：目的探讨miR-338-3p对白介素(IL)-13诱导的人支气管上皮细胞系(BEAS-2B)损伤和卵清蛋白(OVA)诱导的哮喘小鼠气道炎性反应的影响。方法OVA复制哮喘小鼠,分为对照组(control)、模型组(model)、miR-NC agomir和miR-338-3p agomir干预组;HE染色观察肺组织病理形态;TUNEL染色观察肺组织中细胞凋亡;ELISA检测肺组织中炎性因子IL-1β和肿瘤坏死因子(TNF)-α水平。用IL-13诱导BEAS-2B细胞损伤,分为对照组、IL-13组、IL-13+miR-NC和IL-13+miR-338-3p mimic组;MTT法检测细胞活性;流式细胞测量术检测细胞凋亡;ELISA检测细胞中IL-1β和TNF-α水平。生物信息学、荧光素酶法、Western blot和功能修复实验检测miR-338-3p与Ras同源基因(Rho)的靶向关系。结果与模型组比较,miR-338-3p agomir干预组小鼠肺组织的炎性细胞浸润和气道壁增厚等病理表现明显减轻,肺组织中细胞凋亡及IL-1β和TNF-α水平均降低(P<0.05)。与对照组比较,IL-13+miR-338-3p mimic组BEAS-2B细胞活性增高(P<0.05),细胞凋亡及细胞中IL-1β和TNF-α水平均降低(P<0.05)。Rho为miR-338-3p靶基因,过表达Rho减轻了miR-338-3p mimic对IL-13诱导的BEAS-2B细胞损伤和细胞炎性反应治疗作用。结论上调miR-338-3p可靶向Rho基因抑制哮喘相关气道炎性反应和肺上皮细胞损伤。Objective To investigate the effects of miR-338-3p on interleukin(IL)-13-induced human bronchial epithelial cell line(BEAS-2B)injury and airway inflammation in mice with ovalbumin(OVA)-induced asthma.Methods OVA was used to replicate an asthma model of mice,which were divided into control group,model group,miR-NC agomir and miR-338-3p agomir intervention groups.HE staining microscopy was employed to observe the pathological morphology of lung tissue,while TUNEL staining was used to assess cell apoptosis in lung tissue.ELISA was conducted to measure the levels of interleukin-1β(IL-1β)and tumor necrosis factor(TNF)-αin lung tissue.The BEAS-2B cells were subjected to IL-13-induced injury and divided into control group,IL-13 group,IL-13+miR-NC group,and IL-13+miR-338-3p mimic group.Cell viability was assessed with MTT assay.Flow cytometry was employed to evaluate cell apoptosis.The level of IL-1β and TNF-α in cells was measured by ELISA.The targeting relationship between miR-338-3p and Ras homologous(Rho)was investigated using bioinformatics analysis,luciferase assay,Western blot,and functional repair assay.Results Compared to the model group,the miR-338-3p agamid intervention group exhibited a significant reduction in inflammatory cell infiltration and airway wall thickening in lung tissue,as well as decreased cell apoptosis and the level of IL-1βand TNF-α in lung tissue(P<0.05).Compared to the control group,cell viability of BEAS-2B cells in the IL-13+miR-338-3p mimic group exhibited a significant increase(P<0.05),while apoptosis and level of IL-1β and TNF-α within the cells demonstrated a notable decrease(P<0.05).Rho was a target gene of miR-338-3p,and over-expression of Rho attenuated the effect of miR-338-3p mimic on IL-13-induced injury and inflammation in BEAS-2B cells.Conclusions Up-regulation of miR-338-3p can inhibit asthma-related airway inflammation and injury of lung epithelial cells with a potential mechanism targeting at Rho gene.

关 键 词：哮喘 miR-338-3p 气道炎性反应 Ras同源基因(Rho) 

分 类 号：R966[医药卫生—药理学]