中国组织工程研究

中国组织工程研究 ›› 2010, Vol. 14 ›› Issue (32): 5897-5902.doi: 10.3969/j.issn.1673-8225.2010.32.002

• 骨髓干细胞 bone marrow stem cells • 上一篇    下一篇

骨髓间充质干细胞对阿霉素肾病大鼠肾脏nephrin表达的影响

董 晨，杨焕丹，丰炳峰，关凤军，赵 彤   

  1. 徐州医学院附属医院儿科，江苏省徐州市  221000
  • 出版日期:2010-08-06 发布日期:2010-08-06
  • 作者简介:董晨★，女，1965年生，江苏省徐州市人，汉族，1999年徐州医学院毕业，硕士，主任医师，主要从事小儿肾脏疾病诊断及治疗的研究。 dc2260@163. com
  • 基金资助:

    江苏省教育厅自然科学基金(08KJB320018)。

Effects of bone marrow mesenchymal stem cells on nephrin expression in the kidney of adriamycin-induced nephritic rats

Dong Chen, Yang Huan-dan, Feng Bing-feng, Guan Feng-jun, Zhao Tong   

  1. Department of Pediatrics, Hospital Affiliated to Xuzhou Medical College, Xuzhou  221000, Jiangsu Province, China
  • Online:2010-08-06 Published:2010-08-06
  • About author:Dong Chen★, Master, Chief physician, Department of Pediatrics, Hospital Affiliated to Xuzhou Medical College, Xuzhou 221000, Jiangsu Province, China dc2260@163.com
  • Supported by:

    the Natural Science Foundation of Department of Education of Jiangsu Province,No 08KJB320018*

PDF

354

被引次数

22

摘要：

背景：目前对于骨髓间充质干细胞移植治疗急性肾损伤的研究较多，但对治疗慢性肾脏疾病的研究报道甚少。
目的：观察经腹主动脉近肾动脉处移植的骨髓间充质干细胞对阿霉素肾病大鼠足细胞的修复作用及对肾脏Nephrin mRNA表达的影响。
方法：全骨髓法+贴壁筛选法分离培养纯化SD大鼠骨髓间充质干细胞用于移植。SD大鼠随机数字表法分为3组：正常组为正常SD大鼠，无任何干预措施；模型组大鼠尾静脉1次注射阿霉素5 mg/kg+当日腹主动脉注入生理盐水1 mL；骨髓间充质干细胞治疗组大鼠尾静脉1次注射阿霉素5 mg/kg+当日腹主动脉注入干细胞1×107 个，体积约1 mL。各组于处理后7，14，28 d 3个时间点各取8只大鼠用于取材。流式细胞仪对骨髓间充质干细胞表面分子进行鉴定，磺基水杨酸-硫酸钠比浊法测24 h尿蛋白，全自动生化分析仪测血生化指标(白蛋白、血脂)，苏木精-伊红染色及透视电镜下观察肾组织的病理变化，RT-PCR检测肾组织Nephrin mRNA的表达。
结果与结论： ①与正常组相比，模型组大鼠均出现肾病综合征表现，以腹水、大量蛋白尿、低蛋白血症、高胆固醇血症及足细胞足突广泛融合为特征；骨髓间充质干细胞治疗组较模型组则有明显改善。②与正常组相比，模型组大鼠肾组织Nephrin mRNA表达明显降低(P < 0.05)；骨髓间充质干细胞治疗组与模型组相比，肾组织Nephrin mRNA表达有所回升(P < 0.05)。提示骨髓间充质干细胞移植能明显增加阿霉素肾病大鼠肾组织nephrin mRNA的表达，并对受损的肾脏组织起到一定的保护作用。

关键词: 肾病综合征, 骨髓间充质干细胞, 移植, nephrin, 阿霉素, 蛋白尿, 大鼠

Abstract:

BACKGROUND: Numerous studies have addressed bone marrow mesenchymal stem cells (BMSCs) transplantation for treating acute kidney injury, but few reports have concerned BMSCs transplantation for chronic kidney disease.
OBJECTIVE: To investigate the repair effects of BMSCs transplantation on podocytes of adriamycin-induced nephrotic rats via abdominal aorta and effects on Nephrin mRNA expression in the kidney.
METHODS: BMSCs of Sprague Dawley rats were harvested, cultured and purified by whole bone marrow method + adherence screening method, and then used for transplantation. The Sprague Dawley rats were randomly assigned to three groups: normal group: normal Sprague Dawley rats, no intervention; model group: injection of adriamycin once into caudal vein 5 mg/kg + infusion of saline 1 mL into abdominal aorta on the same day; BMSCs group: injection of adriamycin once into caudal vein      5 mg/kg + infusion of stem cells 1×107 into abdominal aorta on the same day, about 1 mL volume. Eight rats from each group were collected at 7, 14 and 28 days following treatment. The cell membrane antigens were detected with flow cytometry analysis. The urine protein was measured at 24 hours using sulfosalicylic acid-sodium sulfate turbidimetric method. Serum albumin and blood fat were examined utilizing automatic biochemistry analyzer. The renal histological changes were observed under transmission electron microscope and using hematoxylin-eosin staining. The expression of Nephrin mRNA in the kidney tissues was measured using reverse transcription-polymerase chain reaction.
RESULTS AND CONCLUSION: Compared with normal group, rats in model group developed nephropathy characterized by ascites, proteinuria, hypoalbuminemia, hypercholesteremia, and progressive renal injury. However, the symptoms in BMSCs group were significantly ameliorated compared with model group. Nephrin mRNA expression in kidney was significantly decreased in the model group compared with normal group (P < 0.05). Compared with model group, Nephrin mRNA expression was increased in BMSCs group (P < 0.05). Results have suggested that BMSCs play a protective effect in ardriamycin-induced nephrotic rats via increasing nephrin mRNA expression in kidney tissue.

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