摘要
目的探讨促红细胞生成素(EPO)预处理骨髓间充质干细胞(BMSC)用于预防大鼠肾移植术后急性排斥反应的作用。方法将BMSC分为5组,分别为对照组(不含EPO)、A组(EPO终浓度10 IU/m L)、B组(EPO终浓度100 IU/m L)、C组(EPO终浓度500 IU/m L)和D组(EPO终浓度1 000 IU/m L),每组细胞培养24 h和48 h,采用3-(4,5-二甲基噻唑-2)-2,5-二苯基四氮唑溴盐(MTT)法测定BMSC的细胞增殖率。将BMSC分为2组,分别为BMSC组(不含EPO)和EPO-BMSC组(EPO终浓度500 IU/m L),每组细胞培养48 h后,采用蛋白印迹(Western blot)法检测各组BMSC表面趋化因子受体(CXCR)4的蛋白表达水平。以Wistar大鼠为供体,SD大鼠为受体,建立肾移植急性排斥反应模型后,将受体鼠随机分为4组,每组6只,分别为对照组(未进行任何干预)、EPO组(术后立即经尾静脉注射1 m L含有500 IU的EPO溶液)、BMSC组(术后立即经尾静脉注射1 m L的1×106/m L BMSC细胞液)、EPO-BMSC组(经尾静脉注射1 m L500 IU/m L EPO体外培养的含有1×106/m L BMSC细胞液),采用血清肌酐(Scr)试剂盒测定血标本中Scr水平,采用Western blot法检测移植肾组织中干扰素(IFN)-γ和白细胞介素(IL)-4的蛋白表达水平。结果培养24 h,各组BMSC增殖率比较,差异均无统计学意义(均为P>0.05)。培养48 h,与对照组比较,C组(EPO终浓度500 IU/m L)的BMSC增殖率较高,差异有统计学意义(P<0.05)。与BMSC组比较,EPO-BMSC组BMSC表面CXCR4的蛋白表达水平更高(P<0.05)。肾移植术后1 d,各组受体鼠的Scr水平比较,差异均无统计学意义(均为P>0.05);术后5 d,与对照组比较,EPO组、BMSC组和EPO-BMSC组受体鼠的Scr水平均明显降低,差异均有统计学意义(均为P<0.05),且EPO-BMSC组受体鼠的Scr水平低于EPO组、BMSC组,差异均有统计学意义(均为P<0.05)。术后1 d和术后5 d,各组受体鼠移植肾组织中IL-4的蛋白表达水平比较,差异均无统计学意义(均为P>0.05)。术后1 d,与对照组比较,EPO组、BMSC组及EPO-BM
Objective To investigate the role of bone marrow mesenchymal stem cells（BMSCs） pretreated with erythropoietin（EPO） in the prevention of acute rejection after renal transplantation in rats. Methods BMSCs were divided into five groups： control group（without EPO）, group A（pretreated with EPO at a final concentration of 10 IU/m L）, group B（pretreated with EPO at a final concentration of 100 IU/m L）, group C（pretreated with EPO at a final concentration of 500 IU/m L） and group D（pretreated with EPO at a final concentration of 1 000 IU/m L）. In each group, the BMSCs were cultured for 24 h and 48 h. The proliferation rate of the BMSCs was determined by 3-（4,5-dimethyl-2-thiazolyl）-2,5-diphenyl-2-H-tetrazolium bromide（MTT） assay. The BMSCs were divided into two groups： BMSC group（without EPO） and EPO-BMSC group（pretreated with EPO at a final concentration of 500 IU/m L）. After 48 h culture, Western blot was adopted to measure the expression level of CXC chemokine receptor（CXCR） 4 protein in BMSCs. Wistar rats were used as the donors, and SD rats were utilized as the recipients to establish the rat models with acute rejection after renal transplantation. The recipient rats were randomly divided into four groups（n=6 in each group） including the control group（without any intervention）, EPO group（injection of 1 m L of solution containing 500 IU EPO via tail vein immediately after surgery）, BMSC group（injection of 1 m L of solution containing 1×106/m L BMSCs via tail vein immediately after surgery） and EPO-BMSC group（injection of 1 m L of solution containing 1×106/m L BMSCs cultured in vitro with 500 IU/m L EPO via tail vein）. The level of serum creatinine（Scr） level was determined by Scr detection kit. Western blot was used to detect the expression levels of interferon（IFN）-γ and interleukin（IL）-4 proteins. Results After 24 h culture, the proliferation rate of BMSCs did not significantly differ among all groups（all P〈0.05）. After 48
作者
张亚
周松
刘永光
范礼佩
李民
郭颖
赵明
Zhang Ya;Zhou Song;Liu Yongguang;Fan Lipei;Li Min;Guo Ying;Zhao Ming(Department of Organ Transplantation,Zhujiang Hospital,Southern Medical University,Guangzhou 510280,China)
出处
《器官移植》
CAS
CSCD
北大核心
2018年第4期283-289,共7页
Organ Transplantation
基金
广东省重大科技专项(2015B020226005)
广东省省级科技计划项目(2016A020220014)
