间充质干细胞条件培养基对糖尿病肾病大鼠肾脏的保护作用

摘要

Objective To determine the protective effect of conditioned medium from mesenchymal stem cells ( MSCs ) on the diabetic nephropathy (DN) rats after intraperitoneal osmotic mini-pump implantation.Methods Thirty Sprasue-Dawley (SD) male rats were randomly divided into 3 groups, these are, normal control group, DN model group and MSCs conditioned medium group , with 10 rats in each group .The rats of the normal control group were fed common food , while those in DN group and MSCs group were fed a high-fat high-sucrose diet.The MSCs group was implanted with Alzet osmotic pump after DN models were confirmed.Blood glucose, serum insulin, triglycerides and other indices were tested in the 3 groups of rats.Renal structural damage was evaluated by periodic acid-Schiff (PAS) staining.Expression of nephrin in the renal tissues was detected by Western blotting .Results The blood glucose, serum insulin, 24-hour urinary protein , serum creatinine and blood urea nitrogen were significantly higher in the MSCs group than in the DN group (P<0.05), and the glomerular mesangial area and the glomerular volume were obviously decreased (P<0.05).The MSCs group had significantly decreased expression of nephrin protein than the normal control group (P<0.05), but the level was higher in comparison with that of the DN group (P<0.05).Conclusion Continuous pumping of conditioned medium from mesenchymal stem cells into abdominal cavity exerts a protective effect on the renal function of DN rats .%目的：利用微量渗透泵持续恒量地向糖尿病肾病（DN）模型大鼠腹腔内泵入间充质干细胞（MSCs）条件培养基，观察其对DN大鼠的保护作用。方法将30只SD大鼠随机分为3组：正常组，DN模型组和MSCs条件培养基组，每组10只。正常组给予普通饲料喂养；DN模型组大鼠每天给予高脂高糖饲料；MSCs条件培养基组是对造模成功的DN大鼠植入Alzet微量渗透泵。检测血糖、血清胰岛素和甘油三酯等指标，观察肾脏病理情况，免疫印迹法（ Western blot ）检测肾组织nephrin蛋白的表达情况。结果与DN模型组大鼠相比，MSCs条件培养基组的血糖、血清胰岛素、24 h尿蛋白、血肌酐和血尿素氮均显著增高，差异具有统计学意义（P＜0.05）。 MSCs条件培养基组大鼠的肾小球系膜截面积与肾小球体积均显著减小（P＜0.05）。与正常组大鼠相比，MSCs条件培养基组大鼠nephrin蛋白的表达情况显著降低（ P＜0.05），但与DN模型组相比显著增加（P＜0.05）。结论微量渗透泵持续泵入MSCs条件培养基可显著改善DN大鼠肾功能。