脂肪间充质干细胞移植减轻大鼠肾缺血再灌注损伤的机制  

Underlying mechanism by which adipose-derived mesenchymal stem cell transplantation alleviates renal ischemia-reperfusion injury in rats

作　　者：雷雨 刘蓉安[2] 曾帆[2] Lei Yu;Liu Rongan;Zeng Fan(Southwest Medical University, Luzhou 646000, Sichuan Province, China;Department of ICU, SichuanProvincial People’s Hospital, Chengdu 610072, Sichuan Province, China)

机构地区：[1]西南医科大学,四川省泸州市646000 [2]四川省人民医院重症医学科,四川省成都市610072

出　　处：《中国组织工程研究》2019年第5期749-755,共7页Chinese Journal of Tissue Engineering Research

基　　金：四川省干保部课题(2015-204);项目负责人:曾帆~~

摘　　要：背景:缺血再灌注引起的急性肾损伤是有较高死亡率的临床常见问题,尚缺乏有效治疗方法。目前,已有一些间充质干细胞移植应用于改善此病情的研究,但可能机制仍不明确。目的:观察脂肪间充质干细胞移植对大鼠肾缺血再灌注损伤的保护作用及其可能机制。方法:实验所用SD大鼠购自北京维通利华实验动物技术有限公司,分离培养SD大鼠腹股沟部位脂肪组织,获得脂肪间充质干细胞并鉴定;将30只大鼠夹闭左肾动脉,45 min后去除动脉夹恢复血流建立肾缺血再灌注大鼠模型,随机分为移植对照组和干细胞移植组,每组15只,另取15只大鼠作为假手术组,其中,移植对照组尾静脉注射生理盐水,干细胞移植组尾静脉注射脂肪间充质干细胞。在移植后5,15和30 d进行下列检测:荧光显微镜观察大鼠肾组织冰冻切片中脂肪间充质干细胞定植情况;ELISA法检测血清肌酐与尿素氮水平;生化法检测肾组织中丙二醛含量;石蜡切片苏木精-伊红染色观察肾组织病理学结构并进行肾小管损伤评分;Western blot法检测肾组织中谷胱甘肽过氧化物酶4的表达。结果与结论:(1)分离培养的SD大鼠脂肪间充质干细胞表达CD90和CD44,具有多向分化能力,移植对照组肾组织无绿色荧光,干细胞移植组在移植后5 d可见绿色荧光;(2)与移植对照组比较,干细胞移植组血清肌酐与尿素氮水平、肾组织丙二醛含量及肾小管损伤评分降低(P <0.05),肾组织谷胱甘肽过氧化物酶4表达升高(P <0.05),肾组织形态结构有明显改善;(3)以上结果说明,脂肪间充质干细胞移植可减轻大鼠肾缺血再灌注损伤,可能通过促进肾组织中谷胱甘肽过氧化物酶4的表达发挥抗脂质过氧化作用。BACKGROUND:Acute renal injury induced by ischemia and reperfusion is a common clinical problem with high mortality,and there is no effective treatment.Mesenchymal stem cell transplantation has been used to improve the condition of the disease,but the possible mechanism is still unclear.OBJECTIVE:To investigate the protective effect of adipose-derived mesenchymal stem cell transplantation on renal ischemia-reperfusion injury in rats and its possible mechanism.METHODS:Adipose-derived mesenchymal stem cells were isolated from the inguinal adipose tissue of Sprague-Dawley rats(provided by Beijing Vital River Laboratory Animal Technology Co.,Ltd.in China),cultured and identified.Thirty rat models of renal ischemia-reperfusion were established by45minutes of clipping the left renal artery and randomly divided into transplantation control group and stem cell transplantation group,15rats in each group.Another15rats were selected as sham operation group.Adipose-derived mesenchymal stem cells were injected into the tail vein of the stem cell transplantation group and the normal saline was injected into the tail vein of the control group.The following tests were performed5,15and30days after transplantation:frozen sections of rat kidney tissue were observed by fluorescence microscope for the colonization of adipose-derived mesenchymal stem cells;serum creatinine and urea nitrogen levels were detected by ELISA;malondialdehyde content in the renal tissue was detected by biochemical method;renal pathological structure was observed by hematoxylin-eosin staining in paraffin slices and renal tubule damage was scored;glutathione peroxidase4expression in renal tissue was detected by western blot.RESULTS AND CONCLUSION:Adipose-derived mesenchymal stem cells from Sprague-Dawley rats expressed CD90and CD44,and showed multidirectional differentiation.There was no green fluorescence in the renal tissue of transplantation control group,but green fluorescence was observed in the stem cell transplantation group at5days after transplantation

关 键 词：脂肪间充质干细胞 肾缺血再灌注损伤 干细胞移植 谷胱甘肽过氧化物酶4 肾小管损伤 急性肾损伤 组织工程 干细胞 间质干细胞 

分 类 号：R456[医药卫生—治疗学] R364[医药卫生—临床医学]