摘要
目的探讨骨髓间充质干细胞(bMSCs)移植对单侧输尿管结扎(UUO)模型大鼠肾小管上皮细胞凋亡的影响。方法自Wistar大鼠股骨和胫骨分离bMSCs,经体外传代培养、成脂诱导及鉴定后制备细胞悬液。54只Wistar大鼠随机分为假手术对照组(n=18)、bMSC注射组(bMSC组,n=18)和生理盐水注射组(PS组,n=18)。bMSC组和PS组大鼠于UUO模型建立的同时经下腔静脉分别注入bMSCs和等体积生理盐水。于建模后第3、7和14天分批处死大鼠并采集肾脏组织标本,RT-PCR技术检测肾小管细胞FasLmRNA表达,免疫组化染色检测FasL蛋白表达及分布;TUNEL法检测肾小管细胞凋亡,计算凋亡指数。结果与假手术组比较,bMSC组和PS组建模后各时间点FasL mRNA及蛋白表达均明显上调,肾小管细胞凋亡指数显著增加(均P<0.05)。建模后第3、7天时,bMSC组FasL mRNA和蛋白表达的上调幅度及肾小管细胞凋亡指数的增加程度均明显低于PS组(P<0.05);至建模后第14天时,bMSC组与PS组各项检测指标比较,差异均无统计学意义(P>0.05)。结论静脉注入bMSCs对UUO模型大鼠肾小管上皮细胞凋亡有短期抑制作用。
Objective To investigate the effects of bone marrow cell apoptosis in rats with unilateral ureter obstruction （UUO）. mesenchymal stem cells （bMSCs） on tubular epithelial Methods MSCs were isolated from femur and tibia of male Wistar rats, and were cultured and passaged in vitro. Cell suspension was prepared after identification of differentiation of bMSCs into adipocytes. Fifty-four rats were randomly divided into sham operation group （ n = 18） , hMSCs injection group （bMSC group, n = 18） and normal saline infusion group （ PS group, n = 18）. BMSCs were injected by the inferior vena cava in bMSC group when UUO model was set up, and normal saline of equal volume was injected in PS group. Rats were killed and renal tissues were harversted on the third, seventh and fourteenth day after model establishment. The mRNA expression of FasL was measured by RT-PCR, the protein expression was detected by immunohistochemical staining, the cell apoptosis was determined by TUNEL method, and the apoptotic index （AI） was calculated. Results Compared with sham operation group, the mRNA and protein expression of FasL and AI in bMSC group and PS group were significantly increased at each time point （P 〈 0.05）. Compared with PS group, the mRNA and protein expression of FasL and AI in bMSC group were significantly decreased on the third and seventh day after model establishment （P 〈 0.05）. There was no significant difference in each parameter on the fourteenth day between bMSC group and PS group （P 〉 0.05）. Conclusion Intravenous injection of MSCs can inhibit the apoptosis of renal tubular epithelial cells in rats with UUO for a short time.
出处
《上海交通大学学报（医学版）》
CAS
CSCD
北大核心
2009年第4期421-425,共5页
Journal of Shanghai Jiao tong University:Medical Science
基金
上海交通大学医学院基金(06XJ21005)~~
