【作者】 章璐；

【导师】 曹勇；

【作者基本信息】 暨南大学 ， 中西医结合临床， 2009， 硕士

【摘要】 目的:探讨中药健脾疏肝解毒方对DEN诱发性大鼠肝癌的作用和机制,研究脾虚肝郁刺激对肝癌形成的影响。以期为临床运用健脾疏肝解毒方治疗肝癌特别是预防和控制肝癌形成提供客观的依据。方法:健康72只SD雄性大鼠,体重110士20g,正常饲养三天,称重后按SPSS随机分为DEN对照组,DEN中药高剂量组,DEN中药低剂量组;病证结合模型组,病证结合中药高剂量组,病证结合中药低剂量组。DEN水溶液按照浓度为1mlDEN:10000ml灭菌蒸馏水的比例配成,给予分组后各组大鼠自由饮用,每天饮用,连续22周,建立诱发性肝癌大鼠实验模型。病证结合模型各组大鼠采用先连续灌胃生大黄煎液,并慢性束缚大鼠四肢,限制活动和断食以施加脾虚肝郁刺激因子,同时给予DEN溶液自由饮用,建立脾虚肝郁大鼠肝癌病证模型。DEN中药高、低剂量组和病证结合中药高、低剂量组,分别按30mg/kg、10mg/kg给予中药,DEN对照组和病证模型组给予生理盐水,连续22周。实验结束后第2天,各组大鼠于乙醚麻醉下处死,观察肝脏表面病变结节,称取肝脏、脾脏重量,计算肝脏指数,脾指数。HE染色,光镜下观察大鼠肝脏组织结构,并运用免疫组化法检测肝脏组织突变型P53,Survivin的表达水平。结果:大鼠经脾虚肝郁因素刺激和DEN作用下的病证结合模型组,与单纯DEN诱发性大鼠肝癌对照组比较,免疫器官的重量有显著下降,二者有显著性差异(P＜0.05);而健脾疏肝解毒方能显著提高诱癌大鼠免疫器官的重量,各组大鼠肝指数、脾指数结果与DEN对照组相比,DEN中药高、低剂量组有显著统计学意义(P＜0.01),中药高剂量组脾指数显著高于中药低剂量组(P＜0.01);中药高、低剂量组肝指数低于DEN对照组(P＜0.01);病证中药高、低剂量组脾指数均高于病证模型组,尤其是病证中药高剂量组更为显著(P＜0.01);病证中药高、低剂量组肝指数均低于病证模型组(P＜0.01)。健脾疏肝解毒方亦能降低肝癌组织突变型P53,Survivin蛋白的表达量,与DEN对照组相比,DEN中药高、低剂量组有明显差异(P＜0.01);与病证结合模型组相比,病证中药高、低剂量组均有统计学意义(P＜0.05),病证结合中药高剂量组差异显著(P＜0.01)。结论:健脾疏肝解毒方能明显抑制大鼠肝癌的形成,并能增强肝癌大鼠免疫器官的重量,其作用机制可能与健脾疏肝解毒方下调肝癌组织突变P53,Survivin基因表达以及增强肝癌大鼠的免疫功能有关。并且脾虚肝郁刺激对二乙基亚硝胺诱发大鼠肝癌的的形成有促进作用,对免疫功能有明显抑制作用。更多还原

【Abstract】 Objective:to observe The Effects of Jianpi Shugan Jiedu Recip on the Expression of p53 and survivin gene During DEN-Induced Hepatocarcinogenesis in Ra,and further explore its mechanism,with a view to effective clinical use of the JPSGJD recipe detoxification of liver cancer,especially cancer of advanced treatment provide an objective basis.Methods:male healthy rat weighting 110±10g,(n=72).feeding 3 days in normal. Later,rat were randomly divided into 6 groups by SPSS:DEN Control group, Middle-dose of JPSGJD with DEN group,High dose of JPSGJD with DEN group; BINZHENGJIEH modle Control group of JPSGJD,Middle dose of JPSGJD group, High dose of JPSGJD group.the chemotherapy drug was given by intraperitoneal injection for all group to the end.DEN SolutionConcentration is 1mlDEN:10000ml pured Sterile water,the rat drank that Solution everyday,until the end of the experiment.To establishe the DEN-Induced Hepatocarcinogenesis in Rat model. Secondly,we give traditional Chinese medicine DAHUANGConcentrated liquid Ig for BINZHENGJIEH modle Control group’s all rat,at the same time,Long-term constraint rat limbs,Restraint rat’ food,activtation to anger them.Throught this way to establish PIXUGANYU model on Rat.the dosage of Middle dose of JPSGJD group and High dose of JPSGJD group isl0mg/kg,30mg/kg.At the end of the experiments,the rat were killed by cervical dislocation,we weighed the tumor,the spleen,the liver.Calculated the rate of inhibitory effect of liver carcinoma and the index of the spleen,the liver.HE stain,light microscopy level of metastasis.We measured P53、Survivin expression of the rat through Immunohistochemistry technique method.To explore the mechanism of the JPSGJD recipe on the liver cancer of mechanism.Results:It has showed that the JPSGJD recipe has remarkably suppressing effects on cancer of the DEN-Induced Hepatocarcinogenesis in Rat.It can significantly increase the inhibition rate of DEN-Induced Hepatocarcinogenesis in rat of liver cancer(compared with the control group,P＜0.01);In addition,the JPSGJD recipe can significantly improve the mice’s immunologic function,especially for High dose of the SGJPJD recipe(compared with the control group,P＜0.01).The JPSGJD recipe can decrease the expression of P53 and Survivin protein expression(compared with the control group,P＜0.01),and the JPSGJD recipe with DEN group which compared with the DEN group and model group could have statistic differences(P＜0.01).Conclusion:The JPSGJD recipe can inhibit liver cancer in rat,Their role may be related to P53、Survivin gene reduce expression;The JPSGJD recipe can also enviously enhance the immunologic function of liver cancer rat.更多还原