miR-21对肾脏缺血/缺氧耐受中血管适应性调整的调控作用  

The Role of miR-21 in Renal Ischemia/hypoxic Tolerance by Regulation of Vascular Function

作　　者：郭漫 沈道琪 於佳炜[1] 徐辰祺 林静[1] 宋娜娜 耿雪梅[1] 许佳瑞[1] 丁小强[1] 徐夏莲[1] GUO Man;SHEN Daoqi;YU Jiawei(Department of Nephrology,Zhongshan Hospital,Fudan University,Shanghai Medical Center of Kidney,Shanghai Institute of Kidney Disease and Dialysis,Shanghai Key Laboratory of Kidney Disease and Blood Purification,Shanghai,200032)

机构地区：[1]复旦大学附属中山医院肾脏科,上海市肾脏疾病临床医学中心,上海市肾病与透析研究所,上海市肾脏疾病与血液净化重点实验室,上海200032

出　　处：《中国中西医结合肾病杂志》2023年第9期768-772,I0001,共6页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：国家自然科学基金青年科学基金资助项目(No.82000706);国家自然科学基金面上项目(No.81770734);上海市肾脏疾病临床医学中心建设项目(No.2017ZZ01015)

摘　　要：目的:探讨微小RNA 21(miR-21)通过调节血管扩张,减轻肾脏缺血/再灌注(ischemia/reperfusion,I/R)损伤的作用机制。方法:通过夹闭小鼠双侧肾蒂35 min后恢复灌注构建肾脏I/R模型,术前给予小鼠腹腔注射氯化钴(cobalt chloride,CoCl_(2)),分为CoCl_(2)干预组和生理盐水对照组。两组均在再灌注后24 h处死小鼠,观察肾功能(血清肌酐)、肾组织病理评分(HE染色)、miR-21和其靶基因PTEN、pAKT/AKT和内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)蛋白表达的变化。体外给予人血管内皮细胞CoCl_(2)干预。通过锁核苷酸修饰的anti-miR-21抑制miR-21表达,观察对miR-21及下游通路PTEN/AKT/eNOS蛋白表达的影响。结果:CoCl_(2)可显著减轻I/R小鼠的肾功能和组织病理损伤,并诱导I/R小鼠肾脏和血管内皮细胞miR-21高表达,抑制PTEN、激活AKT磷酸化并上调eNOS蛋白表达,肾脏一氧化氮(nitric oxide,NO)含量增加。抑制血管内皮细胞miR-21表达,细胞PTEN蛋白上调,磷酸化AKT和eNOS蛋白的表达降低,上清液NO含量减少。结论:miR-21可能通过抑制靶基因PTEN调节AKT/eNOS通路,扩张肾脏内血管,从而对肾脏I/R损伤起到保护作用。Objective:To identify the mechanism of microRNA 21(miR-21)on alleviating renal ischemia/reperfusion(I/R)injury by regulating vascular dilatation.Methods:Reperfusion was restored after clamping bilateral renal pedicles of mice for 35min to construct I/R model.Cobalt chloride(CoCl_(2))was intraperitoneally injected before operation.The mice were randomly divided into CoCl_(2)pretreatment group and saline control group.After reperfusion for 24 h,all mice were sacrificed.Renal function(serum creatinine),renal histopathological score(HE staining),miR-21 and the protein of PTEN/pAKT/AKT pathway and endothelial nitric oxide synthase(eNOS)(real-time PCR and western blot)were estimated.In vitro study,human umbilical vein endothelial cells were treated with CoCl_(2).Locked nucleotide acid modified anti-miR-21 was used to inhibit the expression of miR-21.The expression of miR-21 and protein of PTEN/AKT/eNOS pathway were detected.Results:CoCl_(2)alleviated renal I/R injury,inducing expression of miR-21 in vivo and vitro study,downregulating the expression of PTEN protein,upregulating the level of phosphorylated AKT and eNOS protein,and increasing the level of nitric oxide(NO)in kidneys.Inhibiting the expression of miR-21 in vascular endothelial cells increased the expression of PTEN protein,decreased the level of phosphorylated AKT,eNOS protein and NO in cellular supernatant.Conclusion:miR-21 could protect renal against I/R injury by dilating renal vascular through the PTEN/AKT/eNOS pathway.

关 键 词：缺血/再灌注 MIR-21 血管内皮细胞 

分 类 号：R692[医药卫生—泌尿科学]