microRNA-33a对分化骨髓间充质干细胞移植治疗大鼠肾缺血-再灌注损伤的影响  

Effect of microRNA-33a on the transplantation of differentiated bone marrow mesenchymal stem cells for renal ischemia-reperfusion injury

作　　者：许明伟 孙晓松[1] 石洪波[1] 曾东扬 XU Mingwei;SUN Xiaosong;SHI Hongbo;ZENG Dongyang(Department of Urology,Xiangyang Central Hospital,Affiliated Hospital of Hubei University of Arts and Science,Xiangyang 441021,China)

机构地区：[1]襄阳市中心医院湖北文理学院附属医院泌尿外科,441021

出　　处：《现代泌尿生殖肿瘤杂志》2022年第5期302-307,共6页Journal of Contemporary Urologic and Reproductive Oncology

摘　　要：目的探寻microRNA-33a(miRNA-33a)对分化骨髓间充质干细胞(BMSCs)移植治疗大鼠肾缺血-再灌注损伤(RIRI)的作用和机制。方法以传代并分化的大鼠BMSCs为上皮细胞,通过转染miRNA-33a mimics/inhibitors/阴性对照来测定BMSCs的生长曲线。将转染miRNA-33a mimic/inhibitor的BMSCs作为干预措施,输注至RIRI动物模型,建模手术后24 h抽取大鼠尾静脉血,测定血清肌酐和尿素氮含量以及超氧化物歧化酶、过氧化氢酶、谷胱甘肽过氧化物酶等抗氧化酶含量。处死大鼠后获取肾脏标本,分别行HE和Masson染色观察各组肾脏病理组织学的形态差异,另对肾组织蜡块行增殖细胞核抗原(PCNA)免疫组化染色,记录阳性细胞的分化增殖情况。结果将转染了miRNA-33a mimic/inhibitor的BMSCs输注至RIRI动物模型后,miRNA-33a通过加快BMSCs氧化应激进程、损伤肾功能,抑制miRNA-33a则可减缓BMSCs的氧化应激进程并保护肾功能;HE及Masson染色结果显示,miRNA-33a可刺激肾小管扩张、增加炎性细胞浸润,并加速肾脏组织纤维化及胶原沉积,抑制miRNA-33a表达对于缓解肾小管上皮细胞肿胀有一定程度的效果;PCNA免疫组化染色结果显示,予以BMSCs治疗后,PCNA阳性细胞计数显著升高,转染miRNA-33a后可显著降低PCNA阳性细胞计数。结论移植分化的BMSCs,对于RIRI存在确切的修复作用,且上述修复作用受到miRNA-33a调控;抑制miRNA-33a表达可降低氧化应激损伤,进而发挥对肾小管上皮细胞的保护作用。Objective To explore the effect and mechanism of miRNA-33a on the transplantation of differentiated bone marrow mesenchymal stem cells(BMSCs)for renal ischemia-reperfusion injury(RIRI).Methods Passage and differentiated BMSCs were treated as epithelial cells,by transfecting miRNA-33a mimics/inhibitors/negative control,to determine the growth curve of BMSCs.The rat RIRI animal model was established.BMSCs transfected with miRNA-33a mimic/inhibitor were used as an intervention measure to injected into the RIRI animal model.The tail vein blood of rats was taken and the contents of serum creatinine,urea nitrogen and antioxidant enzymes such as superoxide dismutase,catalase and glutathione peroxidase were measured at 24 hours after operation.The rats were killed and the renal specimens were stained with HE and Masson to observe the morphological differences of renal histopathology in each group.In addition,the wax blocks of renal tissue were stained with proliferating cell nuclear antigen(PCNA)immunohistochemistry to record the differentiation and proliferation of positive cells.Results After infusing BMSCs cells transfected with miRNA-33a mimic/inhibitor into an RIRI animal model,miRNA-33a can aggravate the oxidative stress process of BMSCs and damage the kidney function.Inhibiting miRNA-33a can relieve the oxidative stress process of BMSCs and protect the kidney features.HE and Masson staining suggested:miRNA-33a can stimulate renal tubule dilatation,increase inflammatory cell infiltration,accelerate renal tissue fibrosis and collagen deposition,and inhibiting miRNA-33a can relieve the swelling of tubular epithelial cells to a certain extent.PCNA immunohistochemical staining suggested:after treatment with BMSCs,the count of PCNA-positive cells increased significantly,and the count of PCNA-positive cells significantly decreased after miRNA-33a transfection.Conclusions Transplantation of differentiated BMSCs cells,there is a definite repair effect on RIRI,and the above-mentioned repair effect is regulated by miRNA-33a.

关 键 词：miRNA-33a 肾缺血-再灌注损伤 骨髓间充质干细胞 

分 类 号：R457.7[医药卫生—治疗学]