摘要
目的检测儿童支气管哮喘肺泡灌洗液(BALF)中miR-130a-3p与自噬相关蛋白7(ATG 7)的靶向调节关系,进一步探讨miR-130a-3p靶向调节ATG 7对炎症因子和气道重塑相关因子的影响。方法收集2018年1月—2021年8月145例接受治疗的儿童支气管哮喘的临床资料,将患儿分为轻度哮喘组(n=45)、中度哮喘组(n=58)和重度及危重度哮喘组(n=42),并收集同时期健康儿童40例为对照组。比较组间miR-130a-3p、ATG 7、炎症因子和气道重塑相关因子的水平。双荧光素酶报告基因实验预测miR-130a-3p与ATG 7的靶向关系。采用转染技术敲低支气管上皮细胞16HBE中ATG 7的水平,并检测其对炎症因子和气道重塑相关因子的影响。结果重度及危重度哮喘组的miR-130a-3p、干扰素-γ(IFN-γ)水平低于对照组、轻度哮喘组和中度哮喘组;重度及危重度哮喘组的ATG 7 mRNA、白细胞介素-4(IL-4)、免疫球蛋白E(IgE)、IL-5和IL-13水平高于对照组、轻度哮喘组和中度哮喘组,组间比较差别均有统计学意义(P<0.05)。重度及危重度哮喘组的Ⅰ型胶原蛋白mRNA、内皮素-1(ET-1)、基质金属蛋白酶2(MMP-2)、MMP-9和转化生长因子-β1(TGF-β1)水平高于对照组、轻度哮喘组和中度哮喘组,组间比较差别均有统计学意义(P<0.05)。miR-130a-3p与ATG 7存在靶向结合位点。敲低ATG 7的表达后IFN-γ水平增加,ATG 7、Ⅰ型胶原蛋白的mRNA、IL-4、IgE、IL-5、IL-13、ET-1、MMP-2、MMP-9和TGF-β1的表达水平降低,组间比较差别均有统计学意义(P<0.05)。结论在儿童支气管哮喘中,miR-130a-3p可能通过靶向调节ATG 7的水平,调节辅助性T细胞1/辅助性T细胞2平衡、减弱气道炎症反应和抑制气道重塑,为儿童支气管哮喘靶向治疗提供方向。
Objective To detect the targeting regulation relationship between miR-130a-3p and autophagy-related protein 7(ATG 7)in children's bronchial asthma bronchoalveolar lavage fluid(BALF),and to further explore the effect of miR-130a-3p on inflammatory factors and airway remodeling by targeting ATG 7 to regulate influence of related factors.Methods A collection of 145 children with bronchial asthma from January 2018 to August 2021,and the children were divided into mild asthma group(n=45),moderate asthma group(n=58)and severe and critically severe asthma group(n=42),and collected 40 healthy children in the same period as the control group.The levels of miR-130a-3p,ATG 7,inflammatory factors and airway remodeling-related factors between the groups were compared.The dual luciferase reporter gene experiment predicted the targeting relationship between miR-130a-3p and ATG 7.The transfection technique knocked down the levels of ATG 7 in bronchial epithelial cells 16HBE,and detected the effects of inflammatory factors and airway remodeling related factors.Results The levels of miR-130a-3p and interferon-γ(IFN-γ)in the severe and critically severe asthma group were lower than those in the control group,mild asthma group and moderate asthma group;while the levels of ATG 7 mRNA,interleukin-4(IL-4),immunoglobulin E(IgE),interleukin-5(IL-5),and interleukin-13(IL-13)in the severe and critically severe asthma group were higher than those in the control group,the mild asthma group and the moderate asthma group,the comparison between the groups was statistically significant(P<0.05).The levels of type I collagen mRNA,endothelin-1(ET-1),matrix metalloproteinase 2(MMP-2),MMP-9 and transforming growth factor-β1(TGF-β1)in the severe asthma group were higher than those in the control group,mild asthma group and moderate asthma group,and there were statistical differences between the groups(P<0.05).There was a targeted binding site between miR-130a-3p and ATG 7.After knocking down the expression level of ATG 7,the level of IFN-γincreased,and the levels of ATG 7 mRNA,type I collagen mRNA,IL-4,IgE,IL-5,IL-13,ET-1,MMP-2,MMP-9,and TGF-β1 were decreased,and there were statistical differences between the groups(P<0.05).Conclusion In children with bronchial asthma,miR-130a-3p may target the level of ATG 7 to regulate T helper 1/T helper 2 cell balance,reduce airway inflammation,and inhibit airway remodeling,providing a direction for bronchial targeted therapy in children.
作者
薛庆辉
XUE Qinghui(The Third Department of Pediatrics,Nanyang First People's Hospital,Nanyang 473200,China)
出处
《福建医科大学学报》
2023年第4期252-257,共6页
Journal of Fujian Medical University
