温肾醒脾化浊方对慢性肾功能衰竭模型肾损伤干预机制的研究

Study on Effect of Wenshenxingpihuazhuo Formula on Kidney Damage in the Rat Model with Chronic Renal Failure

目的探索温肾醒脾化浊方治疗慢性肾衰竭的效应机制,为温肾醒脾化浊法临床应用于治疗慢性肾炎提供科学基础。方法通过阿霉素(6 mg/kg)尾静脉注射构建肾功能衰竭模型大鼠,随机分为正常组、模型组、温肾醒脾化浊方低、中、高剂量组(1.49 g/kg、2.98 g/kg和5.95 g/kg),灌胃给药27 d后,考马斯亮蓝法检测24 h蛋白尿含量;全自动生化分析仪测定各组大鼠血清肾功能肌酐(Scr)、尿素氮(BUN)和尿酸(UA)含量;苏木精-伊红(H&E)染色观察大鼠肾脏病理损伤程度;荧光定量PCR法检测肾组织MMP-9基因、金属蛋白酶抑制剂(TIMP-1)、Col4a2R、转化生长因子-β1(TGF-β1)、纤维连接蛋白1(Fn1)mRNA的表达水平。结果温肾醒脾化浊方显著减少24 h蛋白尿和肾体质量的异常增加,有效降低慢性肾功能衰竭大鼠尿素氮、肌酐、尿酸水平(P<0.05或P<0.01),改善肾组织的炎性细胞浸润、纤维化增生以及肾小球变形萎缩等病变。温肾醒脾化浊方干预显著下调TIMP-1、TGF-β1、Col4a2R和Fn1 mRNA表达,上调MMP-9转录表达(P<0.05),调节MMP-9/TIMP-1失衡(P<0.05)。结论温肾醒脾化浊方可能通过调控MMP9/TIMP-1失衡,下调TGF-β1及Fn1的表达,减少细胞外基质的积聚,从而保护肾功能和延缓肾纤维化。

Objective To explore the molecular mechanism of Wenshenxingpihuazhuo formula on delaying progression of chronic renal failure(CRF),and provide theoretical base in treating chronic renal failure.Methods An rat model of CRF were induced using a single intravenous vein injection with adriamycin(6 mg/kg),the rat treated with adriamycin were divided into five groups:normal group,model group,lowdose group,middle dose group and high dose group.After administration of Wenshenxingpihuazhuo formula,the coomassie brilliant blue method was used to assess 24 h urinary protein;level of uric acid,urea,and creatinine in serum were detected using an automatic biochemical analyzer;kidney histopathological changes were measured by H&E staining.The mRNA expression in renal fibrosis-related signaling(TIMP-1,MMP-9,TGF-β1,Col4a2R and Fn1)was examined by q-PCR analysis.Results Oral administration of Wenshenxingpihuazhuo formula markedly decrease the 24 h urinary protein and renal index,significantly reduced the level of uric acid,urea,and creatinine from serum,obviously alleviated the renal pathological damage,including infiltration of inflammatory cell into kidney tissue,hyperplasia of fibrous tissue and glomerular deformation and atrophy.Furthermore,the mRNA expression of TIMP-1,TGF-β1,Col4a2R,Fn1 in Wenshenxingpihuazhuo formula treated group were decreased as compared to model group(P<0.05 or P<0.01),while MMP-9 transcriptional expression was upregulated(P<0.05).Conclusion Wenshenxingpihuazhuo formula could delay renal fibro and progression of chronic renal failure via regulating transcriptional expression in kidney fibrosis-related signaling,including recover the balance MMP9 and TIMP-1 and downregulation of TGF-β1,Col4a2R and Fn1.