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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(CAAN)大鼠肾脏肝细胞生长因子(HGF)表达的影响。方法雌性Wistar大鼠30只,随机分为移植组、非移植组、正常对照组,每组10只。移植组和非移植组每只大鼠经关木通水煎剂灌胃12周,制作CAAN大鼠模型,第12周末,移植组大鼠经尾静脉输入骨髓干细胞1×108个(约0.35 mL),非移植组大鼠经尾静脉输入0.35 mL9 g.L-1盐水。正常对照组大鼠先用饮用水灌胃12周,然后经尾静脉注射0.35 mL的9 g.L-1盐水。第16周末留取3组大鼠血、尿、肾组织标本。应用免疫组织化学、Western blot及RT-PCR法检测CAAN大鼠肾脏中HGF蛋白和HGF mRNA的表达。结果免疫组织化学染色显示,移植组与非移植组肾脏HGF阳性表达均增强,与正常对照组比较差异有统计学意义(Pa<0.01);移植组增强更明显,与非移植组比较差异有统计学意义(P<0.01);RT-PCR检测显示移植组HGF mRNA表达量明显增强,与非移植组比较差异有统计学意义(P<0.01),非移植组HGF表达增强但较移植组减弱,高于正常对照组(Pa<0.01);肾脏Western blot分析显示同样的变化趋势。结论骨髓干细胞移植可提高肾脏HGF蛋白和HGF mRNA的表达,从而减轻CAAN大鼠肾损害。
Objective To observe the effect of bone marrow derived stem cell transplantation on the expression of hepatocyte growth factor (HGF) in chronic aristolochic acid nephropathy (CAAN) rats. Methods Thirty female Wistar rats were randomly divided into transplantation group, non-transplantation group and normal control group, with 10 rats in each group. At the end of the twelfth week, 1 × 108 bone marrow stem cells (about 0.35 mL) were transplanted into the tail vein of rats in the transplantation group and the non-transplantation group. Rats in the non-transplantation group received 0.35 mL of 9 gL-1 saline through the tail vein. Normal control rats were first treated with drinking water for 12 weeks, and then injected via the tail vein 0.35 mL of 9 g.L-1 saline. At the end of the 16th week, three groups of blood, urine and kidney tissue samples were collected. Immunohistochemistry, Western blot and RT-PCR were used to detect the expression of HGF protein and HGF mRNA in CAAN rat kidney. Results Immunohistochemical staining showed that the positive expression of HGF in renal transplantation group was significantly higher than that in normal control group (P <0.01), and the level of HGF in transplantation group was significantly higher than that in non-transplantation group (P <0.01). The expression of HGF mRNA in the transplantation group was significantly higher than that in the non-transplantation group (P <0.01) by RT-PCR. Compared with the non-transplantation group, the expression of HGF mRNA was increased, Higher than the normal control group (Pa <0.01); Western blot analysis of kidney showed the same trend. Conclusion Bone marrow stem cell transplantation can increase the expression of HGF protein and HGF mRNA in kidneys, and thus reduce renal damage in CAAN rats.