【作者】 姜岳；

【导师】 邱模炎；

【作者基本信息】 中国中医科学院 ， 中医内科学， 2007， 硕士

【摘要】 慢性肾衰竭（CRF）是常见的临床综合征，它发生在各种慢性肾脏病的基础上，缓慢地出现肾功能减退而至衰竭。近年来我国慢性肾衰竭的发病率随着社会老龄化，高血压病、糖尿病等疾病的逐年增多，而呈现出明显上升的趋势。慢性肾衰竭属临床难治性疾病，大部分病人最后往往发展为尿毒症而需要依赖透析生存或进行肾移植。因此，在慢性肾衰竭进展至终末期肾衰竭（尿毒症）之前，采取有效的措施延缓其病程进展，具有重要的意义。肾纤维化是各种肾脏疾病发展到慢性肾衰竭的共同途径，包括肾小球硬化和肾小管间质纤维化，是各种肾脏病终末期的基本病理变化。肾纤维化同其他器官纤维化一样一直是世界医学的研究热点问题之一。肾小管间质损伤在肾脏病的进展过程中起重要作用，肾小球病变程度相同的患者，如果肾小管病变程度不同，预后可相差很大。肾间质纤维化是指细胞外基质，主要是Ⅰ型、Ⅲ型胶原的增加和沉积，其主要病理表现是肾小管上皮细胞减少和间质中出现大量肌成纤维细胞。肌成纤维细胞能合成及分泌Ⅰ、Ⅲ型胶原，促进肾间质纤维化的发生，在肾间质纤维化的进程中起重要作用。由于Ⅰ、Ⅲ型胶原纤维的增生是形成肾间质纤维化的重要因素，因此抑制其增生和促进其降解是防治肾间质纤维化的重要途径。临床研究表明，中医药在延缓慢性肾衰竭病程的进展、改善患者预后等方面的作用具有其独特的优势。大量的实验研究也表明，中药具有良好的抗肾纤维化作用。真武汤出自张仲景《伤寒论》，是治疗慢性肾衰竭脾肾阳虚证的主要代表方剂，临床运用较多。本文借助常用的6种中国期刊文献数据库和Pubmed检索系统，并结合手工检索，查出57年来（1949年～2006年）国内期刊发表的有关运用真武汤防治慢性肾衰竭的文献132篇。通过分析57年来真武汤防治慢性肾衰竭的研究文献发现：从50年代以来对真武汤防治慢性肾衰竭的研究一直持续不断，80年代以后研究逐渐增多；研究内容以临床研究为主，有91篇文献，其中运用真武汤加减方研究的有88篇，另有3篇报道采用真武汤原方治疗取得较好的疗效。实验研究报道较少，仅有7篇文献，有以整体动物实验方法观察真武汤加减方改善大鼠慢性肾衰竭模型肾功能的作用，有运用血清药理学方法观察真武汤含药血清对肾小球系膜细胞增殖的影响以及对人胎肾小球系膜细胞外基质成分中纤维连结蛋白、层粘连蛋白和Ⅳ型胶原的作用。但尚未见真武汤原方防治慢性肾衰竭的整体动物实验研究及其抗肾间质纤维化作用的报道。1实验目的采用符合中医脾肾阳虚证候特征的慢性肾衰竭病证结合模型，观察真武汤对实验性慢性肾衰竭大鼠肾脏的保护作用，并初步探讨真武汤抗肾间质纤维化的作用以及对肾间质纤维化大鼠肾脏Ⅰ、Ⅲ型胶原纤维的影响。2材料与方法2．1实验动物Wistar雄性健康大鼠65只，6周龄，清洁级，体重200±20g。2．2实验药物2．2．1造模药物：腺嘌呤购自北京化学试剂公司，进口分装（批号：04022），用前将其加入蒸馏水中配制成20mg／ml的混悬液。2．2．2治疗药物：真武汤的组方和剂量参考全国高等医药院校教材《方剂学》（第5版），即炮附片9g、茯苓9g、白芍9g、白术6g、生姜9g，将此剂量设为正常成人60kg体重的1日剂量，再根据文献的方法进行大鼠用药剂量的换算，以正常人60kg体重用药剂量的3.5倍为小剂量，正常人60kg体重用药剂量的14倍为大剂量。饮片购自北京同仁堂饮片有限责任公司，由中国中医科学院望京医院药剂科制备成含生药量为0.245g／ml的小剂量浓缩液和含生药量为0.98g／ml的大剂量浓缩液。2．2．3阳性对照药物：尿毒清颗粒，由广州康臣药业有限公司生产（批号：200602），根据文献，大鼠使用尿毒清治疗慢性肾衰竭的剂量为3.75g／kg。使用时将尿毒清颗粒以蒸馏水配制成含药量为375mg／ml的溶液。2．3模型制作方法采用“单肾切除加腺嘌呤灌胃法”建立大鼠慢性肾衰竭模型。大鼠行左肾切除术1周后，每天以200mg·kg^-1·d^-1剂量的腺嘌呤混悬液灌胃造模，连续28天。2．4随机分组方法65只大鼠根据体重按随机数字表法随机抽取10只作为正常组，10只作为假手术组，45只行左肾全切术。术中及术后1周死亡大鼠5只，术后1周，将存活的40只大鼠根据体重按随机数字表法随机分为模型组10只、真武汤小剂量组10只、真武汤大剂量组10只、尿毒清组10只。2．5给药方法正常组、假手术组、模型组：于分组当天开始每天灌胃2ml蒸馏水，直至第28天。治疗各组于术后分组当天开始，每天灌胃给药直至第28天。尿毒清组按3.75g·kg^-1·d^-1的剂量给尿毒清溶液，真武汤大剂量组按9.8g·kg^-1·d^-1的剂量给真武汤大剂量浓缩液，真武汤小剂量组按2.45g·kg^-1·d^-1的剂量给真武汤小剂量浓缩液。各组大鼠按每周体重情况，调整给药和蒸馏水的量。2．6观察指标及测定方法2．6．1一般情况：体重、大便、体毛、毛泽、精神状态、活动状况、摄食量、饮水量。2．6．2 24h尿量及24h尿蛋白定量：实验分组后每隔7天将大鼠分置于代谢笼内留取上午9时至次日上午9时的24h尿液，留尿期间禁食不禁水。记录24h尿量，并用考马斯亮蓝法测24h尿蛋白定量。2．6．3血常规和血生化：给药4周后，用10％水合氯醛（0.15ml／100g）腹腔注射麻醉，从股动脉抽取血样。红细胞、血红蛋白，用血常规分析仪测定；血肌酐、尿素氮、血钙、血磷，用全自动生化分析仪测定。2．6．4肾脏大体观察：用精密电子天平称单肾湿重，并计算大鼠单侧肾重／体重×10³的值，肉眼观察肾脏大小、形态、颜色、质地、包膜、皮质、髓质等。2．6．5病理染色方法：肾脏石蜡切片，分别进行苏木素伊红（HE）染色、六胺银（PASM）染色、马松（Masson）染色、天狼星红（Sirius Red）染色、Mallory染色，光镜下观察大鼠肾组织5种染色的病理形态学改变。2．6．6肾小管间质病变半定量分析方法：根据Sergio的半定量分级标准，随机选取HE染色每组镜下的20个视野，进行半定量分析。2．6．7肾脏组织病理学图像分析方法：在每组PASM染色切片包含肾小球的区域，随机选取200倍光镜下10个视野，用MetaMorph图像分析系统对所选取视野中PASM染色肾小球系膜阳性目标光密度值进行测定，分别计算每组PASM染色肾小球系膜阳性目标积分光密度，再进行统计分析，以反映肾小球系膜增生程度。每组Masson染色切片在光镜200倍视野下分别观察10个互不重叠的皮质视野，借助HMIAS—2000高清晰彩色病理图文报告分析系统拍照，计算纤维化的面积（每张切片测量10个视野，取平均值）占整个视野间质面积（排除肾小球、肾小管、血管所占面积）的百分比值。在偏振光显微镜200倍视野下，观察天狼星红染色各组切片10个互不重叠的皮质视野中的Ⅰ和Ⅲ型胶原，并采用OLYMPUS—DP60高清晰彩色病理图象分析系统分析、计算Ⅰ、Ⅲ型胶原占整个间质视野的百分比值。2．7统计学方法：应用SPSS（V 15.0）统计软件进行统计学处理分析。所有计量资料以均数±标准差（（?）±s）表示，不同组间差异采用单因素方差分析（One-way ANOVA），组间两两比较用LSD检验法，方差不齐时采用秩和检验。等级资料的比较采用秩和检验。P值＜0.05认为差异有显著性意义；P值＜0.01认为差异有非常显著性意义。3实验结果3．1一般情况：造模1周后，模型组大鼠开始出现体重下降、生长抑制现象，同时出现体毛干枯发黄不齐、易脱落，畏寒拱背，精神萎靡，活动减少，尾部及耳廓苍白，便稀等脾肾阳虚的症状，各治疗组的体重下降，轻于正常组，但比模型组重，一般情况的表现均较模型组为轻，但不如正常组、假手术组大鼠健康。3．2 24h尿量和24h尿蛋白定量情况：真武汤大剂量组、真武汤小剂量组24h尿量较模型组增加，其中以真武汤大剂量组增加最显著。真武汤大剂量组、真武汤小剂量组24h尿蛋白定量较模型组显著减少，P＜0.05，且以大剂量组作用为最佳。3．3红细胞数、血红蛋白情况：尿毒清组、真武汤大剂量组红细胞数比模型组显著升高，P＜0.05，而治疗各组之间红细胞数比较，P＞0.05，无显著差异；模型组及治疗各组血红蛋白比正常组显著降低，P＜0.05，治疗各组血红蛋白比模型组显著升高，P＜0.05，其中以真武汤大剂量组最明显，但治疗各组之间血红蛋白比较，P＞0.05，无显著差异。3．4血清肌酐、尿素氮、钙、磷情况：治疗各组SCr较模型组显著降低，P＜0.05；尿毒清组、真武汤大剂量组BUN比模型组显著降低，P＜0.05，而真武汤小剂量组BUN较模型组无明显差异；真武汤大剂量组、真武汤小剂量组血钙比模型组升高，P＜0.05，而尿毒清组血钙较模型组无明显差异；治疗各组血磷较模型组显著降低，P＜0.05；其中以真武汤大剂量组的作用最明显。3．5肾脏大体观察情况：模型组右肾明显增大，苍白色，质地坚硬无光泽，皮质明显变薄、皮髓质分界不清，包膜增厚。真武汤大、小剂量组右肾轻度增大，浅褐色，质地较硬无光泽，皮质变薄、皮髓质分界线可辨，包膜增厚。尿毒清组右肾轻度增大，浅褐色，质地较硬无光泽，皮质变薄、皮髓质分界不清，包膜增厚。3．6单侧肾重及单侧肾重／体重×10³情况：大鼠单侧肾重结果显示：①模型组及治疗各组比正常组、假手术组明显增加，P＜0.05；②真武汤大剂量组比模型组显著减轻，P＜0.05，而尿毒清组、真武汤小剂量组与模型组比较，P＞0.05，无显著差异；③真武汤大剂量组比尿毒清组显著减轻，P＜0.05。大鼠单侧肾重／体重×10³结果显示：①模型组及治疗各组比正常组、假手术组明显增加，P＜0.05；②真武汤大剂量组比模型组显著减轻，P＜0.05，而尿毒清组、真武汤小剂量组与模型组比较，P＞0.05，无显著差异；③真武汤大剂量组比尿毒清组显著减轻，P＜0.05。3．7光镜下肾组织病理学情况：正常组和假手术组大鼠的肾小球、肾小管结构正常。模型组大量肾小管上皮细胞凋亡、坏死、脱落或空泡样变性，肾小管高度扩张，肾小管内、肾间质中有大量金黄色的嘌呤代谢产物结晶沉积、可见大量炎细胞浸润，肾间质重度纤维化增生，肾小囊腔扩张，肾小球系膜细胞轻度增生。尿毒清组肾小管轻度扩张、有少量金黄色的嘌呤代谢产物结晶，肾小管上皮细胞轻度空泡样变性，肾间质轻度纤维化，肾间质可见少量炎细胞浸润，肾小囊腔扩张，肾小球系膜细胞轻度增生。真武汤小剂量组肾小管中度扩张，并且有肾小管萎缩，肾小管上皮细胞坏死、脱落，肾小管内、肾间质中有少量金黄色的嘌呤代谢产物结晶沉积，肾间质中度纤维化，肾小囊腔扩张，肾小球系膜细胞轻度增生。真武汤大剂量组有少量肾小管呈中度扩张，肾间质轻度纤维化，肾小管内、肾间质中有少量金黄色的嘌呤代谢产物结晶，肾小囊腔轻度扩张，肾小球系膜细胞增生不明显。3．8肾小管间质病变半定量分级情况：尿毒清组、真武汤小剂量组、真武汤大剂量组的肾小管间质损害较模型组轻，P＜0.01；而真武汤大剂量组与尿毒清组的肾小管间质损害比较接近，P=0.05，且比真武汤小剂量组的肾小管间质损害要轻，P＜0.05。3．9 PASM染色肾小球系膜积分光密度值定量情况：模型组系膜增生最明显，阳性目标积分光密度值最大，与正常组比较，有显著差异，P＜0.05；各治疗组系膜增生轻于模型组，P＜0.05；而真武汤大剂量组、真武汤小剂量组的系膜增生显著轻于尿毒清组，P＜0.05。3．10 Masson染色肾间质纤维化面积情况：模型组的纤维化面积最大，所占肾间质面积的百分比最高，与正常组比较，有显著差异，P＜0.05；真武汤大剂量组、真武汤小剂量组肾间质中纤维化面积显著低于模型组，P＜0.05；而尿毒清组的肾间质中纤维化面积虽然也低于模型组，但无显著差异，P＞0.05，真武汤大剂量组、真武汤小剂量组肾间质中纤维化面积显著低于尿毒清组，P＜0.05。3．11 Sirius red染色Ⅰ和Ⅲ型胶原的分布面积情况：模型组肾间质中Ⅰ、Ⅲ型胶原分布面积较大，与正常组比较，有显著差异，P＜0.05。真武汤大剂量组、真武汤小剂量组、尿毒清组肾间质中Ⅰ型胶原分布面积显著小于模型组，P＜0.05；而真武汤大剂量组、真武汤小剂量组、尿毒清组的肾间质中Ⅲ型胶原分布面积显著小于模型组，P＜0.05。真武汤大剂量组、真武汤小剂量组肾间质中Ⅰ、Ⅲ型胶原分布面积小于尿毒清组，但无显著差异，P＞0.05。4结论4．1真武汤能够改善大鼠整体状况，具有利尿、降低24h尿蛋白量、改善肾功能、改善肾性贫血、改善钙磷代谢等作用；能使肾小管扩张和炎性细胞浸润减轻，而且肾间质纤维化程度降低，Ⅰ、Ⅲ型胶原纤维增生减轻，肾小囊腔代偿性扩张及肾小球系膜增生减轻。可见，真武汤对单侧肾切除后腺嘌呤性慢性衰竭大鼠肾脏的具有一定的保护作用。4．2真武汤大剂量组、真武汤小剂量组肾间质中Ⅰ、Ⅲ型胶原增生明显减少，低于模型组，可见真武汤有抑制Ⅰ、Ⅲ型胶原纤维增生和促进其降解的作用，从而达到抗肾间质纤维化的作用，这可能是真武汤防治肾间质纤维化、延缓慢性肾衰竭发展进程的机制之一。但其进一步的作用机制有待今后深入研究。4．3实验结果显示，真武汤大剂量组对大鼠慢性肾衰竭的疗效优于真武汤小剂量组，但本实验仅设了两个剂量组，尚不能说明真武汤治疗慢性肾衰竭的有效剂量范围和量效关系。真武汤的君药为附子，为有毒中药，今后应进一步开展真武汤的毒理学研究和安全性评价。更多还原

【Abstract】 Chronic renal failure （CRF） is a common clinical syndrome, it occurs on the basis of chronic kidney disease, appeared to slow renal dysfunction and failure. In recent years, the incidence of chronic renal failure in China, along with the aging of society, hypertension disease, diabetes mellitus and other diseases has increased every year. It showed a clear upward trend. Chronic renal failure is a clinical refractory disease. Most patients often end up on the need dialysis for uremia or kidney transplant to survive. Therefore, in chronic renal failure and end-stage renal failure （uremia）, take effective measures to slow down the disease progression that is of great significance. Renal fibrosis is a common approach to chronic renal failure in various kidney diseases. It includes glomerulosclerosis and tubulointerstitial fibrosis, and it is the basic pathological changes of various end-stage renal disease. As other organs fibrosis, renal fibrosis has been one of the hot-spot issues of medical research in the world. In the progress of kidney disease, tubulointerstitial injury has played an important role in the same patients with glomerular lesions. Although the lesions is same, the prognosis can be a big difference. Renal interstitial fibrosis is the extracellular matrix that is mainly deposition and increasing of collagen of typeⅠand typeⅢ. The main pathological changes were reduced on renal tubular epithelial cells and there was substantial interstitial myofibroblasts. Myofibroblasts can synthesis and secretionⅠ,Ⅲcollagen to promote the occurrence of renal interstitial fibrosis, which plays an important role in the process of renal fibrosis.Ⅰ,Ⅲcollagen fibers forming the proliferation is an important factorof renal fibrosis, therefore inhibiting its proliferation and promoting its degradation are both important ways to combat renal interstitial fibrosis. Clinical research has shown that Traditional Chinese Medicine has its unique advantage in the progression of chronic renal failure and to improve the prognosis of patients.Large number of experiments have demonstrated that the Traditional Chinese Medicine hasa good effect on anti-renal fibrosis.Zhenwu decoction comes from the Treatise on Cold-Attack that is written by Zhang Zhongjing, and is the main representative Formula of yang asthenia of Spleen and Kidney in the treatment of CRF. This paper used six types of Chinese Periodical Literature database and pubmed retrieval system, combined with manual retrieval, and identified 57 years ago（1949～2006） journal published 132 article document by the use of Zhenwu decoction prevention of chronic renal failure. After analysising the research literatures of 57 years about Zhenwu decoction to prevention of CRF, found: Zhenwu decoction to the prevention of chronic renal failure has continued since the 1950s, researches gradually increased since the 1980s, and its research content is clinical research primarily. In 91 literatures of clinical research used Zhenwu decoction addition and subtraction side primarily, but also effect of three reports made better by using the original Formula. Experimental study reported less, only 7 literatures. There really have a whole animal experiments method to improve renal function using Zhenwu decoction in rat models of CRF, and have observed serum pharmacology Zhenwu decoction containing serum on glomerular mesangial cell proliferation and the impact on human fetal glomerular mesangial cells linked extracellular matrix protein fibronectin, laminin and typeⅣcollagen role. Has not seen the original Formula of the Zhenwu decoction preventing and controlling CRF in the overall animal experimental research and its the anti-renal interstitial fibrosis were reported.1 ObjectiveUsing model of Traditional Chinese Medicine syndrome of yang asthenia of Spleen and Kidney binding CRF, to observe the protective effect of Zhenwu decoction of CRF in rats, to explore the role of anti-renal fibrosis andⅠ,Ⅲcollagen fibers affected in renal fibrosis of rat.2 Methods2．1 Experimental animals:Only 65 healthy male Wistar rats, six weeks, cleaning and weighing 200±20g.2．2 Experimental drugs2．2．1 Makes the mold drug: adenine purchased from drug companies, distilled water before adding to the preparation of 20mg/ml suspension.2．2．2 Treatment group drugs: Zhenwu decoction’s reference dose group and the national medical institutions of higher education books"Formulaology", that Aconiti Praeparata Radix 9g, Poriacacos wolf 9g, Paeonia lactiflora Pall 9g, Atractylodes macrocephala koida 6g and Zingiber officinale Rosc 9g. 60kg weight as the normal adult dose of this on the 1st dose. According to the document conversion method rat dosage, 60kg weight to normal dosage of 3．5 times smaller dose and 60kg weight 14 times the normal dosage for the high dose. Pieces purchased from Beijing Tongrentang Pieces limited liability company. Wangjing Hospital Pharmacy prepared, which contains a small dose of concentrated liquid volume and 0．245g/ml, 0．98g/ml large dose of raw drug dose concentrated solution.2．2．3 Positive control drug: Niaoduqing particles produced by the Guangzhou Kang medicine Limited, according to the literature, the use of rats in a dose of 3．75g/kg Niaoduqing. Niaoduqing particles will be used for the preparation of 375mg/ml distilled water solution.2．3 Makes the mold method: "single kidney resection with gavage adenine " was establishing a model of CRF in rats. After rats nephrectomy alone on 1 week, 200mg·kg^-1·d^-1 dose of adenine-scale suspension was administered,28 consecutive days.2．4 Stochastic grouping method: 65 rats were elected randomly according to the weight by the random number table, 10 rats were selected randomly just as a normal group, 10 rats as the sham-operated group, 45 rats were left nephrectomy excisioned. Five rats died after surgery 1 week. The survival of 40 rats were randomly divided into weight based on the random number table, model 10, Zhenwu decoction low-dose group 10, Zhenwu decoction high-dose group 10 and Niaoduqing group 10． 2．5 For medicine method:Normal group, sham-operated group and model group : 2 ml distilled water twice a day up to 28 days. In the treatment groups: Niaoduqing group by3．75g·kg^-1·d^-1 dose to Niaoduqing solution, Zhenwu decoction high-dose group by aquaculture 9．8g·kg^-1·d^-1 dose to high dose Zhenwu decoction concentrated solution, and Zhenwu decoction low-dose group by 2．45g·kg^-1·d^-1 dose to the small dose of concentrated liquid Zhenwu decoction, up to 28 days. According to body weight of rats in each group by each week, the volume of distilled water and dose were adjusted.2．6 Observed and measured methods2．6．1 General: weight, feces, hair, the mental state and situation, and Food intake, water intake.2．6．2 24h urine and 24h urinary protein: every seven days after the rats placed in metabolism cages dedicated 9:00 pm to 9:00 am the next day, and took 24-hour urine. During fasting urine can not help but leave the water. Records of 24 urine and using Coomassie Brilliant Blue measured 24 urinary protein.2．6．3 Blood routine and blood biochemistry: After 4 weeks with 10% chloral hydrate （0．15ml/100g） intraperitoneal injection of anesthesia, blood samples taken from the femoral artery. RBC, Hb detected by blood routine analysis apparatus; Serum creatinine, blood urea nitrogen, serum calcium and phosphorus detected by automatic biochemical analyzer.2．6．4 The renal general observation: kidney sophisticated electronic scales called solitary kidney weight, Unilateral calculated rat kidney weight/body weight×10³ value, the naked eye of kidney size, shape, color, texture coating, cortex, medulla other.2．6．5 Pathology staining method: Renal pathology staining of paraffin sections carried on Hemotoxylin and eosin staining, Periodic Acid-Silver Methenamine staining,Masson staining,Sirius red staining, Mallory staining, and they were observed in renal morphological changes.2．6．6 Semi-quantitative grading of tubulointerstitial lesions method: According to Sergio semi-quantitative analysis of semi-quantitative classification standard, HE staining was randomly selected from each of the 20 endoscopic vision for semi-quantitative analysis.2．6．7 Kidney pathology image analysis method: PASM staining pathological image analysis system mesangial integral optical density value of the quantitative comparison, Masson staining area of quantitative comparison of renal interstitial fibrosis. Sirius red staining was observed in polarized light microscopy sections of theⅠandⅢcollagen. Using image analysis and pathological analysis system calculateⅠand typeⅢpercentage of the total value of inter-quality vision.2．7 Statistical methods: SPSS （15．0 V） statistical software was elected for statistical analysis. All measurement data to mean±standard deviation （（?）±s ）. The differences between the different groups of single-factor analysis of variance. Between 2 groups were compared using LSD test, Wilcoxon test was used at variance missing. Wilcoxon test was used to compare the information hierarchy. P<0．05 was considered statistically significant difference vs. P<0．01 that the difference was statistically significant. 3 Results3．1 General: After makes the mold, the model group rats started to weight loss, the growth inhibition, the same time the hair was dry yellow different, easy peeling and chills, Spiritual dispirited, active reduction, tail and ear pale, and other symptoms of syndrome of yang asthenia of Spleen and Kidney.The treatment group decreased weight, less than normal, it was better than to model group. Performance than the general model of light, but not as the sham-operated group and the normal group health rats.3．2 24h urine and 24h urinary protein: Low-dosage Zhenwu decoction and High-dosage Zhenwu decoction group better than model group in 24h urine. The treatment group compared with 24 urine, Zhenwu decoction high-dose group with the most significant increase. Low-dosage Zhenwu decoction and High-dosage Zhenwu decoction group urinary protein excretion was significantly reduced compared with the model group, P<0．05, High-dosage Zhenwu decoction group is the best between them.3．3 RBC, Hb: Red blood cells of High-dosage Zhenwu decoction group and Niaoduqing group were significantly higher than the model group, P<0．05．RBC between the treatment group compared to P>0．05, no significant difference.Model group and the treatment group was significantly lower than the normal hemoglobin, P<0．05．The treatment groups were significantly higher than model group in hemoglobin, P<0．05, with the most obvious High-dosage Zhenwu decoction group. However,the treatment groups between hemoglobin, P>0．05, no significant difference.3．4 Serum creatinine, blood urea nitrogen, calcium, phosphorus: SCr values of the treatment groups were significantly lower than that in the model group, P<0．05．BUN values of High-dosage Zhenwu decoction group and Niaoduqing group were significantly lower than the model group, P<0．05,but Low-dosage Zhenwu decoction group showed no significant difference compared with the model group. Calcium values of High-dosage Zhenwu decoction group and Low-dosage Zhenwu decoction group increased than model group P<0．05,and Niaoduqing group showed no significant difference compared with the model. Phosphorus values of the treated groups were significantly lower than that in the model group, P<0．05, Zhenwu decoction with the most significant effect of high-dose group.3．5 Kidney general observation: The model group: Right kidney increased significantly, pale color, texture hard Matt. Cortex was thinner, corticomedullary unclear boundaries capsule thickening. High-dosage Zhenwu decoction group and Low-dosage Zhenwu decoction group: a small dose Right kidney mildly increased, light brown, Matt texture harder cortical thinning, corticomedullary line to be identified, coating thickness. Niaoduqing group: Right kidney increased slightly, light brown, with hard Matt cortical thinning, corticomedullary unclear boundaries capsule thickening.3．6 Unilateral renal weight:①Model group and the treatment groups has a marked increase than the normal group and sham-operated group, P<0．05;②In Low-dosage Zhenwu decoction group and High-dosage Zhenwu decoction group weight of unilateral renal significantly reduced than the model group P<0．05, and Niaoduqing group compared with the model group, P>0．05, with no statistical significance;③Unilateral renal weight of High-dosage Zhenwu decoction group significantly reduce than Niaoduqing group, P<0．05．Unilateral renal weight/body weight×10³:①the treatment groups has a marked increase than the normal group and sham-operated group, P<0．05．②In Low-dosage Zhenwu decoction group and High-dosage Zhenwu decoction group unilateral renal weight/body weight×10 significantly reduced than the model group P<0．05, and Niaoduqing group compared with the model group, P>0．05, with no statistical significance;③Unilateral renal weight/body weight×10³ of High-dosage Zhenwu decoction group significantly reduce than Niaoduqing group, P <0．05．3．7 Renal tissue pathology by optical microscope: Glomerular and tubular structures was normal in normal and sham-operated rats. Model group: large tubular epithelial cell apoptosis and necrosis or vacuolar degeneration, tubular great expansion of tubular. There are a lot of renal interstitial golden-yellow product of purine metabolism crystal deposition severe renal interstitial fibrosis proliferation. This shows a large number of renal interstitial infiltration of inflammatory cells, renal cysts expansion of mesangial cells mild hyperplasia. Niaoduqing group: tubular mild expansion, a small golden-yellow crystalline product of purine metabolism. Mild renal tubular epithelial cells in vacuolar degeneration, mild interstitial fibrosis, renal interstitial infiltration of inflammatory cells was observed, expansion of renal cysts, mesangial cells mild hyperplasia. Low-dosage Zhenwu decoction group: tubular moderate expansion with tubular atrophy, tubular epithelial cell necrosis, renal interstitial with a small amount of golden-yellow product of purine metabolism crystal deposition, moderate interstitial fibrosis, renal cysts expansion mesangial cells mild hyperplasia. High-dosage Zhenwu decoction group: small tubular showed moderate expansion of renal interstitial fibrosis, and renal interstitial with a small amount of golden-yellow crystalline product of purine metabolism, mild renal cysts expansion. There was a mesangial cells.3．8 Semi-quantitative grading tubulointerstitial lesions: In Niaoduqing group, Low-dosage Zhenwu decoction group and High-dosage Zhenwu decoction group tubulointerstitial damage compared with the model group, P<0．01; tubulointerstitial damage of High-dosage Zhenwu decoction group close to the Niaoduqing group, P = 0．05,and more than tubulointerstitial damage of Low-dosage Zhenwu decoction group to the light, P<0．05．3．9 PASM mesangial integral optical density quantitative model of mesangial proliferative: The most obvious model of mesangial proliferative positive goals largest integrated optical density value, compared with the normal group,a significant difference, P<0．05．Mesangial proliferative of the treatment groups lighter than model group, P<0．05, and mesangial proliferative of Low-dosage Zhenwu decoction group and High-dosage Zhenwu decoction group significantly lighter than Niaoduqing group, P<0．05．3．10 Masson staining fibrosis in the area of renal interstitial: The model group has the largest group fibrosis, renal interstitial area of the highest percentage share, compared with the normal group, a significant difference, P<0．05．The area of renal interstitial fibrosis of Low-dosage Zhenwu decoction group and High-dosage Zhenwu decoction group were significantly lower than the model group, P<0．05 ,and in the Niaoduqing group renal interstitial fibrosis in the area although lower than the model group, but no significant differences, P>0．05．The area of renal interstitial fibrosis of Low-dosage Zhenwu decoction group and High-dosage Zhenwu decoction group were significantly lower than Niaoduqing group, P <0．05．3．11 Percentage distribution area of Sirius red staining of collagenⅠandⅢ:ⅠandⅢcollagen distribution larger in the model group, compared with the normal group, a significant difference, P<0．05．In Low-dosage Zhenwu decoction group and High-dosage Zhenwu decoction group and Niaoduqing group renal interstitial of type I and III collagen were significantly smaller than the size distribution in model group, P<0．05．In Low-dosage Zhenwu decoction group and High-dosage Zhenwu decoction group renal interstitial of typeⅠandⅢdistribution area less than Niaoduqing group. However, no significant difference, P<0．05．4 Conclusion4．1 Zhenwu decoction can improve the overall situation in rats, diuresis, reducing urinary protein of 24 hours and improving renal function. It can ease of renal anemia, regulate calcium metabolism; expansion and inflammatory cell infiltration in renal tubules, and renal interstitial fibrosis lower,Ⅰ,Ⅲcollagen fibers hyperplasia reduced compensatory renal cysts mesangial expansion and reduce proliferation. This shows that Zhenwu decoction has a protective effec in rats of chronic renal failure using adenine after unilateral nephrectomy.4．2 In renal interstitial of Low-dosage Zhenwu decoction group and High-dosage Zhenwu decoction group,Ⅰand typeⅢhyperplasia fall below the model group. Zhenwu decoction shows inhibitⅠ,Ⅲcollagen fibers proliferation and promote their degradation, thus achieving the anti-renal interstitial fibrosis, which may be Zhenwu decoction on renal interstitial fibrosis and chronic renal failure mechanism of the development process. However, the mechanism to be further in future studies.4．3 Experimental results show that Zhenwu decoction high-dose group is better than Zhenwu decoction low-dose group in rats with chronic renal failure.However, the experiment only use two-dose groups. Zhenwu decoction could not explain the treatment of chronic renal failure effective dose and dose-effect relationships. Aconite is the main drug of toxic medicine in Zhenwu decoction, therefore Zhenwu decoction need further toxicological studies and safety evaluations.更多还原