骨髓间充质干细胞来源外泌体对缺血/再灌注损伤大鼠的肾脏保护作用及其机制  

Protective effect of bone marrow mesenchymal stem cell-derived exosomes on rats with renal ischemia/reperfusion injury and its mechanism

作　　者：李露 杜燕[1] 杜林娜 贾盼攀 刘晓茜 LI Lu;DU Yan;DU Linna;JIA Panpan;LIU Xiaoxi(Nephrology Department,The First Affiliated Hospital of Xi’an Medical University,Xi’an 710077,China)

机构地区：[1]西安医学院第一附属医院肾内科,西安710077

出　　处：《山东医药》2022年第33期41-46,共6页Shandong Medical Journal

基　　金：陕西省自然科学基础研究计划(2021JM-497)。

摘　　要：目的观察骨髓间充质干细胞(BMSCs)来源外泌体对缺血/再灌注(I/R)损伤大鼠的肾脏保护作用,并探讨其机制。方法40只雄性SD大鼠随机分成4组:假手术组(Sham组)、缺血/再灌注组(I/R组)、缺血/再灌注+外泌体组(I/R+Exo组)、缺血/再灌注+外泌体转染miR-145-5p拮抗剂组(I/R+Exo antagomir组)。I/R+Exo组大鼠经尾静脉注射外泌体后制备肾脏I/R模型。I/R+Exo antagomir组大鼠经尾静脉注射转染了miR-145-5p拮抗剂的外泌体后制备肾脏I/R模型。I/R组大鼠经尾静脉注射等量的PBS溶液后制备肾脏I/R模型。Sham组大鼠经尾静脉注射等量的PBS溶液后给予切开及关闭腹腔操作,不予血管夹闭。肾组织再灌注24 h后取肾脏组织和颈动脉血,观察肾组织病理变化并进行损伤评分,检测血清血尿素氮(BUN)、血清肌酐(Scr)含量,检测肾组织miR-145-5p表达,测算肾组织细胞凋亡率,检测肾组织细胞凋亡蛋白裂解半胱氨酸蛋白酶-3(cleaved caspase 3)的表达,观察肾组织超氧化物歧化酶(SOD)、丙二醛(MDA)含量和活性氧簇(ROS)生成情况,检测肾组织炎症因子诱导型一氧化氮合酶(iNOS)、白细胞介素-6(IL-6)、肿瘤坏死因子α(TNF-α)及核因子κB(NF-κB)的表达。结果与Sham组相比,I/R组和I/R+Exo antagomir组大鼠肾组织切片显示肾小球坏死和肥大,小管膨胀及铸型形成;与I/R组相比,I/R+Exo组大鼠肾小球坏死和肥大情况显著减轻,小管膨胀及铸型形成明显减少。I/R组和I/R+Exo antagomir组肾组织损伤评分均高于Sham组和I/R+Exo组(P均<0.05),且I/R+Exo组肾组织损伤评分高于Sham组(P<0.05);I/R组和I/R+Exo antagomir组血清BUN、Scr水平均高于Sham组和I/R+Exo组(P均<0.05)。I/R组、I/R+Exo antagomir组大鼠肾组织miR-145-5p相对表达量均低于Sham组和I/R+Exo组(P均<0.05),且Sham组大鼠肾组织miR-145-5p相对表达量低于I/R+Exo组(P<0.05)。I/R组和I/R+Exo antagomir组大鼠肾组织细胞凋亡率及凋亡蛋白cleaved caspaseObjective To investigate the effect of exosomes(Exo)derived from bone marrow mesenchymal stem cells(BMSCs)on rats with renal ischemia/reperfusion(I/R)injury,and to explore the underlying mechanism.Methods Forty male SD rats were randomly divided into four groups:Sham group,I/R group,I/R+Exo group and I/R+Exo antagomir group.The rats in I/R+Exo group were injected with exosomes through the tail vein,and then the renal I/R operation was conducted.The rats in I/R+Exo antagomir group were injected with exosomes transfected with miR-145-5p antagonist through the tail vein,and then the renal I/R operation was conducted.The rats in I/R group were injected with the same amount of phosphate buffer saline(PBS)solution through the tail vein,and then the renal I/R operation was conducted.The rats in Sham group were injected with the same amount of PBS solution through the tail vein,and then the abdominal cavity was opened and closed without vascular clipping.Twenty-four hours after reperfusion,renal tissues and carotid blood were collected,pathological changes of renal tissues were observed,blood urea nitrogen(BUN)and serum creatinine(Scr)were detected,the mRNA expression of miR-145-5p was determined,the apoptosis rate and the cleaved caspase 3 expression were measured,the superoxide dismutase(SOD)activity,malonaldehyde(MDA)content and reactive oxygen species(ROS)generation were ascertained,and the expression levels of inflammatory factors,including inducible nitric oxide synthase(iNOS),interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)and nuclear factor-κB(NF-κB)were detected.Results Compared with the Sham group,the renal tissues of I/R group and I/R+Exo antagomir group showed glomerular necrosis and hypertrophy,with tubule dilation and casting formation.Compared with the I/R group,the renal tissues of the I/R+Exo group showed significantly relieved glomerular necrosis and hypertrophy,and markedly reduced tubule expansion and casting formation.The renal tissue injury scores of the I/R group and I/R+Exo antagomir group w

关 键 词：外泌体 骨髓间充质干细胞 肾损伤 缺血/再灌注 miR-145-5p 氧化应激 炎症反应 

分 类 号：R692.6[医药卫生—泌尿科学]