摘要
目的:观察基质细胞衍生因子-1(SDF-1)在阿霉素慢性肾病大鼠肾脏内的表达变化以及移植异体骨髓间充质干细胞后对SDF-1表达的影响。方法:体外培养SD大鼠骨髓间充质干细胞(MSCs),缺氧培养,采用流式细胞术检测其表面CXCR4表达,用绿色荧光蛋白(GFP)标记MSCs。SD大鼠左侧肾切除后以2.5mg/kg剂量给大鼠尾静脉注射阿霉素,1次/周,连续2次,慢性肾病模型成功后,将成模大鼠随机分为2组:阿霉素慢性肾病模型组(n=8)和MSCs移植组(n=8),另选8只正常大鼠行假手术做正常对照组。于MSCs移植后不同时间点(1,7,14,30d)处死动物,采用Real-time PCR、Western blotting检测大鼠肾脏组织中SDF-1mRNA和蛋白表达的改变情况,并观察MSCs在大鼠肾脏的分布情况以及大鼠肾脏病理改变。结果:阿霉素慢性肾病SD大鼠肾组织内SDF-1表达较对照组明显增加(P<0.05),并随着时间推移而逐渐增加;GFP标记的MSCs移植后1d发现分布在肾小管,之后逐渐增加,14d时最明显;经缺氧培养的异体骨髓MSCs移植后,SDF-1表达较阿霉素慢性肾病模型组降低,在14d有统计学意义(P<0.05)。结论:在阿霉素慢性肾病大鼠肾组织内,SDF-1表达增加可能与骨髓间充质干细胞迁移增加有一定关系。
Objective： To investigate stromal cell derived factor （SDF）-I expression in adriamycin （ADR） induced nephropathy model in rats and the effects of bone marrow mesenchymal stem cells （MSCs） administration on SDF-1 expression in this model. Methods： MSCs were harvested from SD rats, cultured, and expanded in vitro. The levels of CXCR4 in MSCs treated with hy- poxia for 48 h were detected by flow cytometry. Green fluorescent protein （GFP） was adopted to label MSCs. Nephropathy rat models were induced by left nephrectomy plus ADR injection （2.5 mg/kg, once a week for two weeks）. The rats with nephropathy were randomly divided into two groups： adriamycin nephropathic model control groups （ADR, n= 8） and MSCs transplantation groups （MSC, n=8）. Eight rats were served as normal controls with sham operation. The mR- NA and protein levels of SDF-1 in the renal tissue with adriamycin nephrosis at the end of 1 d, 7 d, 24 d and 30 d after treatment were detected by real-time PCR and Western blotting respective- ly. The labeled cells and renal morphology were examined 14 days after cell transplantation. Results： Compared with those respectively in normal controls, both mRNA and protein levels of SDF-1 increased in ADR group （P^0.05） time-dependently. GFP labeled MSCs had been detec- ted in the renal tubules since 1 d after transplantation, and the green fluorescence was gradually increasing till the 14th day. At the end of 14 d after the transplantation, as compared with the ADR group, expression of SDF-1 was significantly decreased （P〈0.05）. Conclusion. Increased expression of SDF-1 in renal tissue of adriamycin nephrosis might be associated with the immigration of MSCs.
出处
《武汉大学学报（医学版）》
CAS
北大核心
2014年第3期338-342,347,共6页
Medical Journal of Wuhan University
基金
云南省科技厅资助项目(编号:2012FB045)
云南省教育厅资助项目(编号:2011Z043)
