微小RNA-142a-3p通过靶向下调Atg16l1表达介导阿霉素肾病的研究  

Research of microRNA-142a-3p mediates adriamycin-induced nephropathy by targeting down-regulation of Atg16l1 expression

作　　者：袁瑞阳 丁海麦[1] 赵星 苏力德 张学明[1] YUAN Rui-yang;DING Hai-mai;ZHAO Xing;SU Li-de;ZHANG Xue-ming(School of Basic Medicine and Forensic Medicine,Baotou Medical College,Baotou 014040,Inner Mongolia Autonomous Region,China)

机构地区：[1]包头医学院基础医学与法医学院,内蒙古自治区包头014040

出　　处：《中国临床药理学杂志》2025年第3期372-375,共4页The Chinese Journal of Clinical Pharmacology

基　　金：内蒙古自治区自然科学基金资助项目(2020MS08015、2020MS08007);内蒙古自治区留学回区人员创新启动支持计划基金资助项目(2021);内蒙古自治区科技创新引导奖励基金资助项目(Z19SQ2-CZT01);包头医学院花蕾计划基金资助项目(HLJH202315)。

摘　　要：目的探究微小RNA-142a-3p(miR-142a-3p)介导阿霉素肾病(AN)的作用机制。方法将BALB/c小鼠随机分为正常组与模型组,模型组通过尾静脉一次性注射10.5 mg·kg^(-1)阿霉素;正常组使用相同方法注射等体积0.9%NaCl。饲养28 d后取其两侧肾组织用于后续实验。用实时荧光定量聚合酶链反应(RT-qPCR)检测肾组织中miR-142a-3p表达情况;用蛋白质印迹法检测肾组织中自噬相关蛋白16样蛋白1(Atg16l1)及炎症相关的NOD样受体热蛋白结构域相关蛋白3(NLRP3)、白细胞介素-1β(IL-1β)和白细胞介素-18(IL-18)蛋白的相对表达水平。将人胚肾HEK293T细胞分为Atg16l1-WT+NC转染组(转染pmirGLO-Atg16l1-WT和NC mimics)和Atg16l1-WT+miRNA-142a-3p转染组(转染pmirGLO-Atg16l1-WT和miR-142a-3p mimics)。用双荧光素酶实验检测miR-142a-3p对Atg16l1的靶向调控。结果正常组与模型组肾组织中miR-142a-3p的相对表达水平分别为1.20±0.27、2.02±0.14;Atg16l1蛋白的相对表达水平分别为0.81±0.13、0.57±0.12;NLRP3蛋白相对表达水平分别为0.49±0.11、0.82±0.14;IL-1β蛋白相对表达水平分别为0.55±0.11、0.84±0.08;IL-18蛋白相对表达水平分别为0.41±0.15、0.74±0.09,模型组上述指标与正常组比较,在统计学上差异均有统计学意义(均P<0.05)。Atg16l1-WT+NC转染组和Atg16l1-WT+miRNA-142a-3p转染组的相对荧光活性分别为1.26±0.07和0.92±0.10,在统计学上差异均有统计学意义(均P<0.05)。结论阿霉素肾病小鼠肾组织中miR-142a-3p下调靶蛋白Atg16l1的表达,激活炎症小体NLRP3通路,促进阿霉素小鼠肾病的进行性发展。Objective To explore effect and mechanism of microRNA 142a-3p(miR-142a-3p)on adriamycin-induced nephropathy(AN).Methods BALB/c mice were randomly divided into normal group and model group.The model group was injected with 10.5 mg·kg^(-1)doxorubicin at a time through the tail vein,and the normal group was injected with equal volume 0.9%NaCl by the same method.After 28 days of feeding,both sides of the kidney tissues were taken for experiment.Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the expression of miR-142a-3p in the renal tissues.The expression levels of autophagy related protein 16-like protein 1(Atg1611)and inflammatory NOD-like receptor heat protein domain associated protein 3(NLRP3),interleukin-1β(IL-1β)and interleukin-18(IL-18)in the renal tissues were detected by Western blotting.Human embryonic kidney 293T cells were divided into Atg1611-WT+NC transfection group(transfected pmirGLOAtg1611-WT and NC mimics)and Atg1611-WT+miRNA-142a-3p group(transfected pmirGLO-Atg1611-WT and miR-142a-3p mimics).Dual luciferase assay was used to detect the targeted regulation of miR-142a-3p on Atg1611.Results The expression levels of miR-142a-3p in kidney tissues of normal group and model group were1.20±0.27 and 2.02±0.14;the expression levels of Atg1611 were 0.81±0.13 and 0.57±0.12,NLRP3 were0.49±0.11 and 0.82±0.14;IL-1βwere 0.55±0.11 and 0.84±0.08;IL-18 levels were 0.41±0.15 and0.74±0.09,respectively,and the above indexes in the model group were significantly different from those in the normal group(all P<0.05).The relative luciferase activity in Atg1611-WT+NC transfection group and Atg1611-WT+miRNA-142a-3p transfection group were 1.26±0.07 and 0.92±0.10,respectively,with statistical significance(all P<0.05).Conclusion MiR-142a-3p down-regulates the expression of target protein Atg1611in renal tissue of adriamycin nephropathy mice,which activates NLRP3 pathway of inflammatory bodies and promotes the progressive development of adriamycin-induced nephropathy.

关 键 词：阿霉素肾病 微小RNA-142a-3p 自噬相关蛋白16样蛋白1 炎症小体 

分 类 号：R97[医药卫生—药品]