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目的观察骨髓源性干细胞移植对慢性马兜铃酸肾病大鼠肾脏中平滑肌肌动蛋白表达的影响,探讨干细胞对肾脏保护作用的可能机制。方法雌性Wistar大鼠30只,随机分为3组:①移植组;②非移植组;③正常对照组,每组10只,移植组和非移植组经关木通水煎剂灌胃12周,制作慢性马兜铃酸肾病(CAAN)大鼠7模型,第12周末,移植组经尾静脉输入骨髓干细胞2×10个/只(1ml/只),非移植组经尾静脉输入1ml生理盐水;正常对照组,先用饮用水灌胃12周,然后经尾静脉注射等量(1ml)生理盐水。第16周末留取血、尿、肾组织标本,检测血尿素氮、肌酐、尿蛋白定量,并应用免疫组化、免疫印迹及RT-PCR方法研究肾脏中平滑肌肌动蛋白(α-SMA)的蛋白和mRNA的表达情况。结果免疫组化染色显示,移植组与非移植组肾脏α-SMA阳性表达与正常组比较均增强(P<0.01),而非移植组与移植组比较增强更明显(P<0.01);RT-PCR检测显示非移植组α-SMAmRNA表达量与移植组比较明显增强(P<0.01),移植组α-SMA的表达亦增强但较非移植组明显减弱,且显著高于正常对照组(P<0.01);肾脏免疫印迹分析显示了同样的变化趋势。结论骨髓干细胞移植可以降低肾脏α-SMA蛋白和mRNA的表达,可能通过抑制α-SMA表达而对CAAN大鼠肾脏起到保护作用。
Objective To investigate the effect of bone marrow-derived stem cell transplantation on the expression of smooth muscle actin in the kidney of chronic aristolochic acid nephropathy rats and to explore the possible mechanism of stem cell protection on the kidney. Methods Thirty female Wistar rats were randomly divided into three groups: ① transplanted group; ② non-transplanted group; ③ normal control group, with 10 rats in each group. The transplanted group and the non-transplanted group were treated with Guanmutong decoction for 12 weeks, Chronic aristolochic acid nephropathy (CAAN) rat model was established. At the end of the twelfth week, 2 × 10 bone marrow stem cells (1ml / mouse) were transplanted into caudal vein in the transplantation group and 1ml normal saline in the non-transplantation group The control group, first with drinking water for 12 weeks, and then injected through the tail vein (1ml) the same amount of saline. At the end of the 16th week, blood, urine and kidney tissue samples were taken for quantification of blood urea nitrogen, creatinine and urinary protein. Immunohistochemistry, Western blotting and RT-PCR were used to detect the expression of α-SMA Protein and mRNA expression. Results Immunohistochemical staining showed that the positive expression of α-SMA in renal transplantation group and non-transplantation group were significantly increased compared with the normal group (P <0.01), while the non-transplantation group and the transplantation group enhanced more significantly (P <0.01) PCR showed that the expression of α-SMA mRNA in non-transplantation group was significantly higher than that in transplantation group (P <0.01), the expression of α-SMA in transplantation group was also increased, but it was significantly lower than that in non-transplantation group (P < 0.01); Western immunoblot analysis showed the same trend. Conclusion Bone marrow stem cell transplantation can reduce the expression of α-SMA protein and mRNA in kidney and may play a protective role in the kidney of CAAN rats by inhibiting the expression of α-SMA.