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目的观察骨髓间充质干细胞对阿霉素肾病大鼠的治疗作用及黏着斑激酶(FAK)的影响。方法建立阿霉素肾病大鼠模型,分别以干细胞与波尼松对照治疗,测定24 h尿蛋白,电镜观察肾组织结构,实时荧光定量PCR检测Nephrin、Desmin、FAK表达水平的变化,Western blot检测肾组织FAK蛋白及磷酸化,免疫组织化学方法检测Nephrin表达水平。结果 (1)模型组大鼠均出现大量蛋白尿及足细胞足突广泛融合,骨髓间充质干细胞治疗组较模型组有改善。(2)与正常组比较,模型组Nephrin表达减少(P<0.05),Desmin mRNA表达增多(P<0.05),FAK mRNA表达升高(P<0.01);与模型组比较,各治疗组Nephrin表达水平增多(P<0.05),Desmin mRNA表达水平减少(P<0.05),FAK mRNA表达减少(P<0.01)。(3)模型、各治疗组Nephrin蛋白表达水平低于正常组(P<0.05),各治疗组Nehprin蛋白表达水平较模型组有所增高(P<0.05)。(4)模型组FAK总蛋白及磷酸化蛋白增高(P<0.01),各治疗组FAK总蛋白及磷酸化蛋白较模型组降低,P<0.05。结论骨髓间充质干细胞移植能抑制FAK活化,减轻蛋白尿,增加阿霉素肾病大鼠肾组织Nephrin表达水平,同时降低Desmin mRNA表达水平,对肾小球损伤起保护作用。
Objective To observe the therapeutic effect of bone marrow mesenchymal stem cells on adriamycin-induced nephropathy rats and the effect of focal adhesion kinase (FAK). Methods The adriamycin-induced nephropathy rat model was established. The control group was treated with stem cell and prednisone respectively. Urinary protein was measured for 24 hours. The renal tissue was observed by electron microscope. The expression of Nephrin, Desmin and FAK was detected by real-time fluorescence quantitative PCR. Renal FAK protein and phosphorylation, immunohistochemical detection of Nephrin expression levels. Results (1) A large number of proteinuria and podocyte foot processes were found in the model group, and the bone marrow mesenchymal stem cells treatment group was improved compared with the model group. (2) Compared with normal group, Nephrin expression in model group decreased (P <0.05), Desmin mRNA expression increased (P <0.05) and FAK mRNA expression increased (P <0.01) (P <0.05). The expression of Desmin mRNA decreased (P <0.05) and FAK mRNA decreased (P <0.01). (3) model, Nephrin protein expression level in each treatment group was lower than that in normal group (P <0.05). Nehprin protein expression level in each treatment group was higher than that in model group (P <0.05). (4) FAK total protein and phosphorylated protein in model group increased (P <0.01), FAK total protein and phosphorylated protein in each treatment group decreased compared with model group (P <0.05). Conclusion Bone marrow mesenchymal stem cells can inhibit the activation of FAK, reduce the proteinuria and increase the expression of Nephrin in renal tissue of Adriamycin-induced nephropathy rats. At the same time, it can decrease the expression of Desmin mRNA and protect glomerular injury.