禾肾丸对阿霉素肾病大鼠蛋白尿及肾组织nephrin蛋白表达影响

目的:研究禾肾丸对阿霉素肾病大鼠蛋白尿及肾组织足细胞裂孔膜蛋白nephrin表达的影响,为其治疗肾病蛋白尿提供实验依据。方法:SPF级雄性Wistar大鼠随机分为模型组,禾肾丸组(1.8 g·kg~(-1)),贝那普利组(1 mg·kg~(-1))及空白组。采用一次性尾静脉注射阿霉素6 mg·kg~(-1)复制阿霉素肾病大鼠模型。造模成功后,禾肾丸组及贝那普利组分别给予相应药液,模型组及空白组给予等体积蒸馏水,1次/d,连续28 d。给药结束后,收集24 h尿液,检测尿蛋白,腹主动脉采血,检测血中白蛋白(albumin,Alb),血肌酐(serum creatine,SCr),血尿素氮(blood urea nitrogen,BUN)。苏木素-伊红(HE)检查肾脏组织形态变化,透射电镜观察肾小球足细胞足突及基底膜改变,免疫组化及蛋白免疫印迹(Western blot)法检测肾组织nephrin表达。结果:经禾肾丸治疗后,24 h蛋白尿较模型组显著下降(P0.01),血清SCr,BUN显著降低(P0.01),Alb显著升高(P0.01)。组织病理学检查显示,禾肾丸组肾小球上皮细胞肿胀明显减轻,部分肾小球体积略增大,系膜细胞及系膜基质仅见轻微增生,管腔内未见小管有蛋白管型。超微结构研究显示,禾肾丸组肾小球基底膜光滑均匀,略见增厚,足突清晰完整、偶见融合。免疫组化结果显示,禾肾丸组nephrin蛋白阳性面积率显著高于模型组(P0.01)。Western blot结果显示,nephrin蛋白表达量较模型组显著升高(P0.01)。结论:禾肾丸可能通过上调nephrin表达水平,修复受损足细胞而减少蛋白尿、保护肾脏。

Effect of Heshen Pill on Urinary Protein and Expression of Nephrin Protein in Rats with Adriamycin-induced Nephropathy

Objective: To investigate the effect of Heshen pill (HSP) on urinary protein and the expression of nephrin protein in rats with adriamycin-induced nephropathy. Method: The rats were randomly divided into four groups, namely the model group, the HSP group (1.8 g·kg^-1), the benazepril group (1 mg·kg^-1) and the control group. Nephropathy rat model was induced through injection of adriamycin (6 mg·kg^-1). After successful modeling, the HSP group and the benazepril group were giventhe corresponding drugs, and the model group and the control group were given equivalent volume of distilled water, 1 times a day, for 28 days. At the end of the experiment, 24 h urine was collected for urine protein detection, and aortic blood was collected for albumin(Alb), serum creatine(SCr), blood urea nitrogen(BUN) detection. The morphological changes in renal tissues were assessed by hematoxylin-eosin (HE) staining, and the changes in foot process and basilar membrane of podocytes were probed under transmission electron microscope (TEM). The nephrin protein was measured by immunohistochemistry and Western blot. Result: After the treatment, the 24 h proteinuria of the HSP group was decreased significantly than that of the model group (P<0.01). SCr, BUN were decreased significantly, and Alb was increased significantly of the HSP group than those of the model group (P<0.01). Histopathological examination revealed that glomerular epithelial cell swelling of the HSP group was reduced significantly, glomerular volume was increased slightly, mesangial cells and matrix showed slight hyperplasia, and no protein cast is found in lumen. Ultrastructural examination showed that the glomerular basement membrane of the HSP group was smooth and even, with slight thickening, and foot process was clear and complete, with occasional fusion. Immunohistochemistry showed that the positive area rate of nephrin protein of the HSP group was significantly superior to that of the model group (P<0.01). Western blot demonstrated that the nephrin protein expression of the HSP group was significantly superior to that of the model group (P<0.01). Conclusion: The inhibitive effects of HSP on proteinuria are achieved by promoting nephrin expression and repairing podocyte damage.