中文摘要：

目的 研究微小RNA-382（miR-382）在肾小管间质纤维化（TIF）发病中的作用及相关机制.方法 通过抑制miR-382的表达,观察其丰度对人近端肾小管上皮细胞（HK2）转分化的影响,并验证miR-382与其预测靶基因热休克蛋白60（HSPD1）的互补配对关系.建立小鼠单侧输尿管梗阻（UUO）模型,经尾静脉注射锁核苷酸（LAN）修饰的anti-miR-382,观察抑制miR-382表达对梗阻侧肾小管间质病变及肾组织硫氧还原蛋白（Trx）水平的影响.收集有和无明显TIF的IgA肾病（IgAN）患者肾穿刺标本各6例,原位杂交和免疫组化法检查TIF程度及抗氧化应激水平,分析其与miR-382/HSPD1表达的相关性.结果 体外荧光素酶报告基因检测和定点突变实验结果证实HSPD1为新的miR-382对应的直接靶基因.TGF-β1诱导HK2细胞发生转分化,与对照组相比,TGF组miR-382表达水平显著上调[（6.54±0.96）比（1.12±0.26）,P＜ 0.05];阻断miR-382可使TGF-β1诱导的上皮转分化部分被逆转.体内实验结果表明,UUO模型组小鼠发生明显TIF病变,与假手术组相比,模型组miR-382丰度明显上调[（6.89±2.47）比（1.00±0.42）,P＜ 0.05],HSPD1和Trx蛋白表达水平明显下调.10 mg/kg LNA-anti-miR-382干预组miR-382表达下调,梗阻侧肾组织TIF程度减轻,HSPD1和Trx的蛋白表达水平上调（均P＜ 0.05）.IgAN患者肾组织检查结果提示,与无纤维化组相比,纤维化组肾组织miR-382丰度明显上调,HSPD1蛋白表达水平则明显降低（均P＜0.05）.结论 miR-382在人类及小鼠TIF发病中起重要作用,HSPD1是miR-382的直接靶基因.HSPD1表达下调导致相应的抗氧化应激能力减弱可能是miR-382参与TIF的机制之一.

英文摘要：

Objective To investigate the roles of microRNA-382 （miR-382） in the pathogenesis of renal tubulointerstitial fibrosis （TIF）.Methods Human kidney epithelial cells （HK2）transfected with miR-382 inhibitor （antagomiR-382） were used to examine the effect of miR-382 abundance on cell polarity,as well as to test the complementary relationship between miR-382 and its predicted target gene heat shock protein 60 （HSPD1）,which was further verified by 3＇-untranslated region luciferase assay and site-directed mutagenesis.The role of miR-382 played in the development of renal interstitial fibrosis and redox regulation was examined in a mouse unilateral ureteral obstruction （UUO） model.Locked nucleic acid （LAN）-modified anti-miR-382 was intravenous delivered via tail vein 30 min prior to UUO,and repeated the dosage 24 h after the surgery.For clinical verification,renal biopsy specimens from 12 IgA nephropathy （IgAN） patients were collected,6 patients with moderate to severe TIF and 6 patients without TIF.The relative abundance of miR-382 and HSPD1 protein was analyzed by using in situ hybridization and immunohistochemistry.Results HSPD1 was confirmed to be a new,direct target gene of miR-382 by in vitro 3＇-untranslated region luciferase assay and sitedirected mutagenesis.The development of epithelial transition in HK2 cells was accompanied with upregulation of miR-382 [（6.54±0.96） vs （1.12±0.26）,P 〈 0.05].Blocking the expression of miR-382 could reversed the progression of epithelial transition partially.In UUO mice the abundance of miR-382 was up-regulated [（6.89 ± 2.47） vs （1.00±0.42）,P 〈 0.01] while HSPD1 and Trx were downregulated compared with the sham group.Down-regulation of miR-382 was associated with significant decrease in TIF,but increase in HSPD1 and thioredoxin protein compared with UUO group [HSPD1：（0.34±0.10） vs （0.14±0.05）;Trx：（0.79±0.18） vs （0.36±0.16）;all P 〈 0.05].The expression of miR-382 was up-regulated and HSPD1 was sign