养阴活血方药拮抗环孢素慢性肾毒性的大鼠体内实验研究  

In vivo study of Yangyin Huoxue recipe on antagonizing chronic nephrotoxicity of ciclosporine in rats

作　　者：黄效维 刘洲 谢敏妍 

机构地区：[1]广州市番禺区中心医院肾内科,511400

出　　处：《器官移植》2012年第6期338-349,358,共13页Organ Transplantation

基　　金：广东省中医药管理局科研课题(100054)

摘　　要：目的探讨养阴活血方药拮抗环孢素(CsA)慢性肾毒性的作用及机制。方法将50只Sprague-Dawley(SD)大鼠随机分为对照组、依那普利组、中药低剂量组、中药中剂量组、中药高剂量组,每组各10只。采用钠耗竭法制作大鼠CsA肾毒性模型:给予钠缺乏饮食,每日皮下注射CsA12.5mg/(kg·d),连续4周。同时,按组别相应予以蒸馏水、依那普利(依那普利10倍成人剂量)、低剂量中药(养阴活血方药5倍成人剂量)、中剂量中药(养阴活血方药10倍成人剂量)、高剂量中药(养阴活血方药20倍成人剂量)灌胃。建组后14d、28d记录各组大鼠死亡率。建组后7d、14d、21d、28d,测定各组大鼠的血清肌酐(Scr)、尿肌酐、24h尿蛋白定量,计算内生肌酐清除率(Ccr)。建组后28d处死大鼠,取右肾组织进行病理学检查和免疫组织化学染色,观察细胞外基质(extracellular matrix,ECM)成分,包括纤维连接蛋白(fibronectin,FN)、胶原蛋白(collagen,Col)Ⅳ和其调节因子转化生长因子(TGF)-β1的表达;取左肾组织进行逆转录聚合酶链反应(RT-PCR),分别检测FN、ColⅣ、TGF-β1,以及抑制ECM生成基因包括组织型纤溶酶原激活物(tissue-type plasminogen activator,tPA)、尿激酶型纤溶酶原激活物(urokinase-plasminogen activator,uPA)、纤溶酶原激活物抑制剂(plasminogen activator inhibitor,PAI)-1基因的核糖核酸(RNA)表达。结果建组后14d,中药高剂量组的死亡率为20%,依那普利组和对照组各为10%,中药中剂量组、中药低剂量组均无死亡。建组后28d,除中药高剂量组的死亡率为20%、中药低剂量组的死亡率为0外,其余3组死亡率均为10%。建组7d后各组大鼠肾功能逐渐恶化,24h尿蛋白定量增加,于建组后14d达到高峰。建组14d后进入CsA慢性肾毒性期,各治疗组大鼠的肾功能均有所改善、24h尿蛋白定量减少,但对照组仍然继续恶化。建组后28d,中药高剂量组和中药中剂量组的Ccr明显高于对照组(均Objective To investigate the effect and mechanism of Yangyin Huoxue recipe on antagonizing chronic renal toxicity of ciclosporine(CsA). Methods Fifty Sprague-Dawley(SD) rats were randomly assigned to 5 groups including the control group, enalapril group, low-dose Chinese herbal drugs group, medium-dose Chinese herbal drugs group and high-dose Chinese herbal drugs group, with 10 rats in each. All rats were given salt-deficiency diet and CsA 12.5 mg/(kg·d)everyday for 4 consecutive weeks to build the rat nephrotoxicity of CsA model. And the rats in the 5 groups were fed with distilled water, the enalapril(10 times of adult dosage), low-dose Chinese herbal drugs(5 times of adult dosage), medium-dose Chinese herbal drugs (10 times of adult dosage) and high-dose Chinese herbal drugs (20 times of adult dosage) by intragastric administration respectively at the same time. The death rates of the rats were recorded after day 14, day 28. The serum creatinine(Scr), urine creatinine, 24-hour urine protein were tested on day 7、day 14、day 21、day 28 and calculated the endogenous creatinine clearance rate(Ccr). The right renal tissues of rats were harvested on day 28. Histopathological examination and immunohistochemistry methods were used to observe the component of extracellular matrix(ECM) including the expression of fibronectin(FN), collagen(Col)Ⅳ and regulatory factor transformation growth factor (TGF)-β1. The left renal tissues were harvested to test the FN, ColⅣ, TGF-β1 and the ribonucleic acid (RNA) expression of the suppressor genes of ECM including the tissue-type plasminogen activator(tPA), urokinase-plasminogen activator(uPA), plasminogen activator inhibitor(PAI)-1 by reverse transcription polymerase chain reaction (RT-PCR)method. Results The death rates of the rats were 20%, 10%, 10% on the day 14 in the group of high-dose Chinese herbal drugs group, enalapril group,control group respectively.And no death occurred in the medium-dose and low-dose Chinese herbal drugs group. The death rates were 20% on da

关 键 词：养阴活血方药 环孢素 药物毒性 肾毒性 慢性 中药 纤维化 

分 类 号：R617[医药卫生—外科学]