复肾功方对CRF大鼠肾脏保护作用的研究

发布时间：2018-03-16 23:26

  本文选题：慢性肾功能衰竭　切入点：复肾功方　出处：《重庆医科大学》2016年硕士论文　论文类型：学位论文

【摘要】：背景:慢性肾功能衰竭(Chronic Renal Failure,CRF)是由先天或后天原因导致的慢性进行性肾实质损害,致使肾脏功能严重受损的临床综合征,主要表现为氮质血症、代谢产物潴留、水电解质紊乱、酸碱平衡失调和全身各系统症状,为肾脏疾病持续发展的最终归宿。据有关统计,全球终末期肾功能衰竭靠透析维持生命者多达360万人,而中国各种肾脏病人有4000余万人,每年新发生的慢性肾功能衰竭患者则有10-30万人左右。有人预测,未来10年世界慢性肾功能衰竭患者治疗费用至少需要1.1万亿美元,如此庞大的费用已使政府感到棘手。中医药防治慢性肾功能衰竭疗效确切,而复肾功方是首届国医大师、著名中医学家、全国名老中医郭子光教授经过60余年来临床验证总结出来的、治疗慢性肾功能衰竭确有卓著疗效的经验方。因此,研究复肾功方对CRF大鼠肾脏的保护作用,可为中医药治疗CRF寻找新的治疗靶点提供实验依据,而本课题研究对于深入挖掘国医大师临床经验具有十分重要的临床实用价值、社会经济价值及科学研究价值。目的:为了研究复肾功方对慢性肾功能衰竭大鼠肾脏的保护作用,本课题检测了慢性肾功能衰竭大鼠肾组织α-SMA、E-cadherin及Nephrin等蛋白的表达,分别从抗肾纤维化、保护肾小管及足细胞等角度,探讨复肾功方保护慢性肾功能衰竭大鼠肾脏的作用机制,以期为开发一个安全有效的治疗crf的中成药提供实验基础。方法:取雄性sd大鼠55只,适应性饲养1周后,随机分为5组: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,其余各组大鼠食用含0.5%腺嘌呤饲料,每只大鼠按1天食入20g料块计算,连续21d。造模21天后,随机抽取各组大鼠1只,摘取眼球采血,分离血清,检测scr和bun等指标以验证造模是否成功。造模成功后,所有大鼠均停用含0.5%腺嘌呤的饲料,改用常规饲料。从造模成功后第1天起,正常组和模型组给予ns20ml?kg-1体重灌胃,复肾功方低、中、高剂量组分别以4、8、16g生药?kg-1体重灌胃复肾功方煎剂,每日1次,连续30d。治疗30d后,末次给药后禁食不禁水12h,按20%乌拉坦5ml?kg-1进行麻醉,摘取眼球采血3-5ml,静置1-2h,4000r?min-1离心10min,分离血清,取上清液,低温4℃保存待检测用。颈椎脱臼处死大鼠,打开腹腔取出肾脏,并做检测前组织的相关处理。he染色和masson染色检测肾组织形态学改变和肾间质纤维化程度,免疫组化及westernblotting检测α-sma及e-cadherin蛋白的表达,免疫荧光及rt-pcr检测nephrinmrna的表达。结果:与正常组比较,模型组大鼠肾组织α-sma表达明显上调(p0.05),e-cadherin及nephrin表达明显下调(p0.05);与模型组比较,复肾功方各剂量组肾组织α-sma表达明显下调(p0.05),e-cadherin及nephrin蛋白表达则显著上调(p0.05)。光镜下正常组大鼠肾小管和肾小球结构正常,未见明显组织学改变;模型组大鼠肾小球变形萎缩,球囊闭塞,肾小管萎缩,管腔明显扩大,间质纤维化明显,间质炎性细胞弥漫浸润。经复肾功方治疗后,复肾功方低剂量组大鼠肾小球、肾小管萎缩及间质纤维化有所减轻,间质炎性细胞弥漫浸润;复肾功方中剂量组大鼠肾小球、肾小管萎缩及间质纤维化明显减轻,间质炎性细胞浸润减轻;复肾功方高剂量组大鼠肾脏病理改变显著减轻,间质炎细胞浸润明显减轻。结论:复肾功方对CRF大鼠肾脏的保护作用,可能通过复肾功方抗肾纤维化、保护肾小管及足细胞等作用来实现的,其机制可能与复肾功方抑制肾组织α-SMA的表达,以及上调肾组织E-cadherin及Nephrin的表达有关。
[Abstract]:Background: chronic renal failure (Chronic Renal Failure, CRF) is caused by congenital or acquired causes chronic progressive renal damage, resulting in serious damage to renal function in clinical syndrome, mainly for azotemia, metabolite retention, electrolyte disturbances, acid-base imbalance and systemic symptoms, as the final incidence of kidney disease development. According to statistics, the global end-stage renal failure on dialysis to sustain life as many as 3 million 600 thousand people, and China all kidney patients more than 4000 people, chronic renal failure patients can occur each year there are about 10-30 million. It is predicted that the next 10 years the world in patients with chronic renal failure treatment costs at least $1 trillion and 100 billion, huge costs have made the government feel difficult. The curative effect of Chinese medicine in treating chronic renal failure and renal function exactly, is the first Chinese Medical University Division, national famous traditional Chinese medicine doctor, Professor Guo Ziguang after 60 years of clinical verification summed up the experience of the treatment of chronic renal failure has outstanding curative effect. Therefore, the research of complex renal function of renal protective effect of CRF rats for the TCM treatment of CRF to find new therapeutic targets for on the basis of experiment, and this research has very important clinical value for the dig Chinese medicine master clinical experience, social and economic value and scientific research value. Objective: To study the complex renal function of chronic renal failure rat renal protective effect, we detected the renal tissue of rats with chronic renal failure alpha -SMA, the expression of E-cadherin and Nephrin protein, respectively, from the anti renal fibrosis and protect renal tubular cells and foot angle, to investigate the chronic renal failure rat kidney complex mechanism of renal protection, in order to 涓哄紑鍙戜竴涓�瀹夊叏鏈夋晥鐨勬不鐤梒rf鐨勪腑鎴愯嵂鎻愪緵瀹為獙鍩虹��.鏂规硶:鍙栭泟鎬�sd澶ч紶55鍙，

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