MiR-21介导丹参多酚酸B在肾脏缺血/再灌注损伤中的保护作用  

MiR-21 Contributes to Salvianolic Acid B-conferred Amelioration of Renal Ischemia/Reperfusion Injury in Mice

作　　者：耿雪梅[1] 吉俊[1] 许佳瑞[1] 陈欣 蒋春波[2] 丁小强[1] 林攀[1] 徐夏莲[1] GENG Xuemei;JI Jun;XU Jiarui(Department of Nephrology,Zhongshan Hospital,Fudan University,Shanghai Institute of Kidney Disease and Dialysis(SIKD),Shanghai Laboratory of Kidney Disease and Dialysis,Shanghai Medical Center of Kidney Disease,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 [2]江苏省苏州市中医医院肾内科,苏州215000

出　　处：《中国中西医结合肾病杂志》2019年第11期950-954,共5页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：中山医院优秀青年人才培养计划(No.2017ZSYXQN07);中山医院青年基金(No.2017ZSQN14);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)介导丹参多酚酸B(salvianolic acid B,Sal B)在肾脏缺血/再灌注(ischemia/reperfusion,I/R)损伤中的保护作用。方法:构建肾脏I/R模型,术前1 h给予小鼠尾静脉注射Sal B,分为四组:(1)假手术组(Sham组)、(2)缺血再灌注组(I/R组)、(3)丹参多酚酸B干预组(SB+I/R组)和(4)生理盐水对照组(NS+I/R组)。四组均在再灌注后24 h处死小鼠,观察Sal B预处理对再灌注后24 h肾功能、肾组织病理评分、细胞凋亡、肾脏miR-21和程序性细胞死亡因子4(programmed cell death 4,PDCD4)表达变化的影响。体外给予人肾小管上皮细胞低氧/复氧和Sal B干预。锁核苷酸(LNA)修饰的anti-miR-21抑制miR-21表达(体外细胞转染),观察miR-21、PDCD4 mRNA和蛋白和细胞凋亡的变化。结果:在体外研究中,Sal B减轻肾小管上皮细胞低氧/复氧损伤,上调低氧/复氧细胞miR-21表达,降低PDCD4蛋白表达(P<0.05)。抑制细胞miR-21表达则显著削弱Sal B的细胞保护作用,anti-miR-21明显上调细胞PDCD4蛋白的表达水平(P<0.01),同时凋亡的细胞比例显著增加(P<0.01)。肾脏I/R小鼠模型中,与I/R组、NS+I/R组相比,SB+I/R组小鼠肾功能和组织病理损伤显著减轻(P<0.01);Sal B诱导再灌注后肾脏miR-21高表达,同时PDCD4蛋白表达有所下降(P<0.01),伴有细胞凋亡显著减少(P<0.05)。结论:Sal B诱导的miR-21通过抑制靶基因PDCD4的表达,减轻肾小管上皮细胞凋亡,从而对肾脏I/R损伤起到保护作用。Objective:To investigate whether microRNA 21(miR-21)contributed to salvianolic acid B(Sal B)-conferred amelioration of renal ischemia/reperfusion(I/R)injury in mice.Methods:On the basis of the mice renal I/R model,Sal B was delivered by intravenous(i.v.tail vein)injection 1h before ischemia.The mice were divided into four groups:(1)Sham group,(2)ischemia/reperfusion group(I/R group),(3)Sal B pretreatment group(SB+I/R group)and(4)saline control group(NS+I/R group).All the mice were sacrificed at 24 h after reperfusion.The effects of Sal B on renal function,renal histopathology score,apoptosis,expression of renal miR-21 and programmed cell death factor 4(PDCD4)at 24 h after reperfusion were detected.In vitro,human renal tubular epithelial cells were treated with Sal B pretreatment and then hypoxia/reoxygenation.Locked nucleotide acid(LNA)modified anti-miR-21 was used to inhibit miR-21 expression(cell transfection in vitro).The expression of miR-21,PDCD4 mRNA and protein and cell apoptosis were estimated.Results:Sal B could protect the renal tubular epithelial cells against hypoxia/reoxygenation injury with up-regulation of miR-21 expression and down-regulation of PDCD4 mRNA and protein(P<0.05).Inhibition of miR-21 expression by LNA anti-miR-21 significantly increased expression of PDCD4(P<0.01)and attenuated the protective effect of Sal B in hypoxia/reoxygenaiton cells,which was shown as significantly increased percentage of apoptotic cell(P<0.01).In mice renal I/R model,compared to the mice from the I/R group and NS+IR group,renal function and pathological damage were attenuated significantly by the Sal B pretreatment(P<0.01).Sal B could induce expression of miR-21 in kidney undergoing I/R insult with the remarkably decreased expression of PDCD4(P<0.01)and cell apoptosis(P<0.05).Conclusion:Sal B-induced miR-21 might decrease apoptosis of renal tubular epithelial cells by inhibiting the expression of target gene PDCD4,thereby contributing to the protection of renal against the I/R injury.

关 键 词：丹参多酚酸B 肾脏缺血/再灌注损伤 MIR-21 细胞凋亡 

分 类 号：R73[医药卫生—肿瘤]