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目的研究复肾功方对慢性肾功能衰竭(chronic renal failure,CRF)大鼠肾组织足细胞裂孔隔膜蛋白(nephrin)mRNA表达的影响,探讨其降低尿蛋白、减轻肾损害的作用机制。方法将55只雄性SD大鼠随机分为正常组,模型组,复肾功方低、中、高剂量组,每组11只。正常组常规饲养,其余4组大鼠食用含腺嘌呤饲料建立慢性肾功能衰竭模型,连续喂养21d。造模成功后,所有大鼠改用常规饲料。正常组和模型组按20mL/(kg·d)灌胃给予生理盐水;复肾功方低、中、高剂量组按大鼠体质量分别以生药4、8、16g/kg灌胃复肾功方,1次/d,连续灌胃30d。实验结束后,收集大鼠尿液测定24h尿蛋白定量,检测各组大鼠血清血肌酐(SCr)和尿素氮(BUN)水平,HE染色检测肾小球组织形态学改变,免疫荧光检测nephrin蛋白表达,Real-Time PCR法观察肾脏nephrin mRNA表达情况。结果与正常组比较,模型组24h尿蛋白定量、SCr和BUN水平升高(P<0.05),nephrin蛋白及mRNA表达下降(P<0.05);与模型组比较,经复肾功方治疗后,24h尿蛋白定量、SCr和BUN水平降低(P<0.05),肾组织nephrin蛋白及mRNA的表达升高(P<0.05)。结论复肾功方降低CRF大鼠尿蛋白,减轻肾损害程度,其机制可能与升高肾组织nephrin蛋白及mRNA的表达、减轻足细胞损伤有关。
Objective To investigate the effects of Fu Shen Keng Geng on expression of nephrin mRNA in the podocytes of rats with chronic renal failure (CRF) and to explore its mechanism of reducing urinary protein and reducing renal damage. Methods Fifty-five male Sprague-Dawley rats were randomly divided into normal group, model group and Fu Shen Gong Fang low, medium and high dose group, 11 rats in each group. The rats in the normal group were fed routinely. The rats in the other four groups were fed with adenine-containing diet to establish the model of chronic renal failure, and were fed continuously for 21 days. After successful modeling, all rats switched to conventional feed. Normal group and model group were given normal saline by intragastric administration of 20mL / (kg · d). Fufang Gongfang low, medium and high dose groups were given intragastric administration of 4,8 and 16g / kg crude drug respectively Side, 1 / d, continuous gavage 30d. At the end of the experiment, urine samples were collected for determination of urinary protein excretion in 24 h. Serum creatinine (SCr) and blood urea nitrogen (BUN) levels were measured in each group. Glomerular morphological changes were detected by HE staining. Nephrin protein The expression of nephrin mRNA in kidney was detected by Real-Time PCR. Results Compared with the normal group, the urinary protein, the levels of SCr and BUN increased (P <0.05) and the protein and mRNA expression of nephrin decreased (P <0.05) in the model group. Compared with the model group, 24h urinary protein, SCr and BUN levels decreased (P <0.05), nephrin protein and mRNA expression increased (P <0.05). Conclusion Fufanggongfang can reduce urinary protein and reduce the degree of renal damage in CRF rats. The mechanism may be related to increasing nephrin protein and mRNA expression in renal tissue and reducing podocyte injury.