微小RNA-21对急性肾损伤模型大鼠线粒体氧化磷酸化的影响及机制研究  

Effect of microRNA-21 on oxidative phosphorylation of mitochondria in rats with acute renal injury and its mechanism

作　　者：张星 高明 赵武 赵成群 康云慧  ZHANG Xing;GAO Ming;ZHAO Wu;ZHAO Chengqun;KANG Yunhui;RASSUL(Department of Nephrology,Xi’an Daxing Hospital,Xi’an 710082,China)

机构地区：[1]西安大兴医院肾内科,陕西西安710082 [2]陕西中医药大学,陕西咸阳712046

出　　处：《陕西医学杂志》2022年第3期274-279,共6页Shaanxi Medical Journal

基　　金：陕西省创新能力支撑计划青年科技新星项目(2021KJXX-56)。

摘　　要：目的:研究微小RNA(miR)-21对急性肾损伤(AKI)大鼠线粒体氧化磷酸化的影响及其相关机制。方法:选取40只雄性SPF级SD大鼠,随机分为对照组、模型组、阴性对照组和miR-21抑制剂组,每组10只。模型组、阴性对照组和miR-21抑制剂组腹腔注射脂多糖(LPS)诱导建立AKI模型,阴性对照组腹腔注射转染阴性对照,miR-21组腹腔注射miR-21 antagomiR。检测各组大鼠的血清尿酸(SUA)、血清肌酐(Scr)、血尿素氮(BUN)。HE染色观察肾组织病理学变化。检测各组线粒体膜电位、ATP含量、氧化磷酸化功能指标;实时荧光定量PCR(qRT-PCR)检测各组miR-21、过氧化物酶体增殖物激活受体γ辅激活因子1α(PGC-1α)、核呼吸因子1(NRF1)和线粒体转录因子A(TFAM)mRNA表达。Western blot测定PGC-1α、NRF1、TFAM蛋白表达。结果:模型组和阴性对照组SUA、Scr和BUN水平高于对照组,miR-21抑制剂组SUA、Scr和BUN水平低于模型组和阴性对照组(均P<0.05)。模型组和阴性对照组线粒体膜电位和ATP含量低于对照组,而miR-21抑制剂组线粒体膜电位和ATP含量高于模型组和阴性对照组(均P<0.05)。HE染色可见模型组和阴性对照组大鼠肾组织存在损伤。模型组和阴性对照组线粒体呼吸功能受到显著抑制,而miR-21抑制剂组线粒体呼吸功能得到改善。模型组和阴性对照组大鼠肾组织中miR-21表达水平高于对照组,而miR-21抑制剂组大鼠肾组织中miR-21表达水平低于模型组和阴性对照组(均P<0.05)。与对照组相比,模型组和阴性对照组PGC-1α、NRF1、TFAM mRNA和蛋白相对表达量较低(均P<0.05);与模型组和阴性对照组相比,miR-21抑制剂组PGC-1α、NRF1、TFAM mRNA和蛋白相对表达量较高(均P<0.05)。结论:miR-21可通过调控线粒体氧化磷酸化关键基因PGC-1α、NRF1和TFAM的表达破坏线粒体氧化磷酸化,诱发大鼠肾损伤。抑制miR-21表达可对线粒体氧化磷酸化功能以及肾组织损伤起到保护作用。Objective:To investigate the effect of microRNA(miR)-21 on oxidative phosphorylation of mitochondria in rats with acute renal injury(AKI)and its mechanism.Methods:40 male SPF grade SD rats were randomly divided into control group,model group,negative control group and miR-21 inhibitor group,with 10 rats in each group.The model group,negative control group and miR-21 inhibitor group were intraperitoneally injected with lipopolysaccharide(LPS)to induce the establishment of AKI model.The negative control group was intraperitoneally injected with transfected negative control,and the miR-21 group was intraperitoneally injected with miR-21 antagomiR.The serum uric acid(SUA),serum creatinine(Scr)and blood urea nitrogen(BUN)were detected.The pathological changes of renal tissue were observed by HE staining.The mitochondrial membrane potential,ATP content and oxidative phosphorylation function of each group were detected.The relative expression of miR-21 and peroxisome proliferator-activated receptorγcoactivator-1α(PGC-1α),nuclear respiratory factor 1(NRF1),mitochondrial transcription factor A(TFAM)mRNA were detected by qRT-PCR.The relative expression of PGC-1α,NRF1,TFAM proteins were detected by Western blot.Results:The levels of SUA,Scr and BUN in model group and negative control group were higher than those in control group,and the levels of those in miR-21 inhibitor group were lower than those in model group and negative control group(all P<0.05).The mitochondrial membrane potential and ATP content in model group and negative control group were lower than those in control group,while those in the miR-21 inhibitor group were higher than those in model group and negative control group(all P<0.05).HE staining showed that the renal tissue of the model group and the negative control group was damaged.The mitochondrial respiratory function was significantly inhibited in the model group and negative control group,while the mitochondrial respiratory function was improved in the miR-21 inhibitor group.The expression level

关 键 词：急性肾损伤 微小RNA-21 线粒体 氧化磷酸化 过氧化物酶体增殖物激活受体γ辅激活因子1α 

分 类 号：R692[医药卫生—泌尿科学]