【作者】 于莉英；

【导师】 王旭东；

【作者基本信息】 南京中医药大学 ， 中医康复学， 2012， 博士

【摘要】 目的：研究中医古典理论“五积”中“肝积”、“脾积”与肝纤维化的相关性,提出“温散”的中医康复法则,在古方基础上创制方药“肝积散”。通过实验研究,观察肝积散对肝纤维化大鼠肝细胞酶、血清肝纤维化标志物、抗脂质过氧化、肝星状细胞及细胞因子的影响,探讨基于温散法的肝积散抗肝纤维化的作用机理。方法：用Wister雄性大鼠102只,适应性饲养1周后随机分为2组,分别为正常对照组12只、造模组90只。造模组腹腔注射40%的四氯化碳石蜡油造模剂,首次0.5ml/100g体重,之后每次为0.3m1/100g体重,每周2次,正常对照组注射等量0.9%氯化钠注射液。进行4周造模之后改为每周1次,再进行8周,共进行12周。造模并以10%的酒精作为以上造模组的唯一饮料,正常对照组给以生理盐水。造模4周之后随机抽取造模组中10只处死,证实肝纤维化模型已经建立成功之后的60只大鼠分为正常对照组、模型组、秋水仙碱片组、肝积散低剂量组、肝积散中剂量组、肝积散高剂量组,并开始进行灌胃给药治疗。具体方法：模型组和正常对照组给予蒸馏水1.5ml/100g体重,秋水仙碱片组按每只0.01mg/100g,肝积散低剂量治疗组给予中药复方0.05g/200g,中剂量治疗组给予中药复方0.1g/200g,高剂量治疗组给予中药复方0.2g/200g,共治疗8周。治疗8周后,眼球后取血后处死大鼠。统一留取大鼠肝右叶标本置于10%甲醛溶液固定,常规石蜡包埋切片,HE染色。结果：根据病理组织切片所示,本研究所有造模大鼠肝脏均可见不同程度的肝细胞水样变性和脂肪变性、纤维结缔组织增生、炎细胞浸润,表明本研究造模成功。模型组大鼠肝组织炎症和纤维化程度较正常组明显增高(P<0.01)；血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、透明质酸(HA)、层粘连蛋白(LN)、Ⅲ型前胶原(PCⅢ)、丙二醛(MDA)、转化生长因子β1(TGF-β1)、肝组织结缔组织生长因子(CTGF)、金属蛋白酶组织抑制因子-1(TIMP-1)、CTGFmRNA、TIMP-1mRNA、肝星状细胞大麻素受体CB1和CB2、活化的肝星状细胞的表达均较正常组明显增高(P<0.05,P<0.01)；血清中超氧化物歧化酶(SOD)较正常组明显降低(P<0.01)。用肝积散高、中、低剂量治疗后,大鼠肝组织炎症和纤维化程度较模型组明显减轻(P<0.05,P<0.01)；血清丙氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)、透明质酸(HA)、层粘连蛋白(LN)、Ⅲ型前胶原(PCIII).丙二醛(MDA)、转化生长因子β1(TGF-β1)、肝组织结缔组织生长因子(CTGF).金属蛋白酶组织抑制因子-1(TIMP-1)、 CTGFmRNA、TIMP-1mRNA、肝星状细胞大麻素受体CB1、活化的肝星状细胞的表达均较模型组明显降低(P<O.05,P<0.01)；血清中超氧化物歧化酶(SOD)的活性和肝星状细胞大麻素受体CB2较模型组明显升高(P<0.01)。结论：基于温散法的肝积散能够下调肝纤维化大鼠血清ALT、AST、HA、LN、PCⅢ、 MDA、TGF-β1、肝组织CTGF、TIMP-1、CTGFmRNA、TIMP-1mRNA.肝星状细胞大麻素受体CB1、活化的肝星状细胞的表达；升高血清SOD和肝星状细胞大麻素受体CB2的表达,这些可能是其抗肝纤维化的部分作用机制。更多还原

【Abstract】 Objective:To study the ancient literature of Chinese medicine with hepatic fibrosis related "Tive ji""hepatic ji"," spleen ji" theory, the principle and prescription are summarized, put forward " warm yang and expell cole" traditional Chinese medicine rehabilitation law, the Cuban side on the basis of creation of prescription" hepatic ji powder". Through the experimental study, observation of hepatic ji powder on hepatic fibrosis of rat liver cell enzymes, serum markers of hepatic fibrosis, lipid peroxidation, hepatic stellate cell and cell factors, based on warm yang and expell cole’s hepatic ji powder on hepatic fibrosis mechanism.Methods:102male Wister rats were raised for1weeks, adaptability and randomly divided into6groups, each group of17. For normal control group, model group, colchicine tablets group, hepatic ji powder low dose group, hepatic ji powder dose group, high dose group of hepatic ji powder.In addition to the normal control group, other groups of intraperitoneal injection of40%carbon tetrachloride paraffin molding agent, the first0.5ml/100g body weight, after each0.3ml/100g body weight,2times a week, normal control group received the same amount of0.9%sodium chloride injection. Zhou Zao dies after4tol times a week. By8weeks, a total of12weeks, molding and by10%alcohol as the above model the only drink. Normal control group with normal saline. Model for4weeks after a random sample of each model in any ldeath, confirmed hepatic fibrosis models have been established successfully started after gavage administration in the treatment of. Specific methods are as follows:the model group and normal control group were given distilled waterl.5ml/100g body weight, colchicine tablets group according to each0.01mg/100g, hepatic ji powder low dose treatment group were treated with compound herb0.05g/200g, hepatic ji powder doses treatment group were treated with compound herb0.1g/200g, hepatic ji powder high dose therapy group was given Chinese medicine compound0.2g/200g, for8weeks.8weeks after treatment, retrobulbar blood rats were sacrificed after. Unified take rat liver right leaf specimens in10%formaldehyde fixed, embedded in paraffin sections stained with HE.Results:According to the pathological tissue section below, all of this research model of rat liver were observed in varying degrees of liver cell hydropic degeneration and fatty degeneration, fibrous connective tissue hyperplasia, infiltration of inflammatory cells, this study demonstrated successfully mould. in the model group rats liver inflammation and fibrosis degree lower than normal group were significantly higher (P<0.01); serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), hyaluronic acid (HA), laminin (LN), type III procollagen (PC III), MDA (MDA), transforming growth factor beta1(TGF beta1), liver tissue connective tissue growth factor (CTGF), tissue inhibitor of metalloproteinase-1(TIMP-1), CTGFmRNA, TIMP-1mRNA, hepatic stellate cell receptors CB1and CB2, activation of hepatic stellate cells expression were higher than normal group increased (P<0.05, P<0.01); serum superoxide dismutase (SOD) compared with the normal group were significantly reduced (P<0.01).Using hepatic ji powder high, medium, low dose treatment, rat liver inflammation and fibrosis degree than those in the model group reduced significantly (P<0.05, P<0.01); serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), hyaluronic acid (HA), laminin (LN), type III procollagen (PC III), malondialdehyde (MDA), transforming growth factor beta1(TGF beta1), liver tissue connective tissue growth factor (CTGF), tissue inhibitor of metalloproteinase-1(TIMP-1), CTGFmRNA, TIMP-1mRNA, hepatic stellate cells, activation of the cannabinoid receptor CB1hepatic stellate cell expression than those in model group were significantly reduced (P<0.05, P<0.01); serum superoxide dismutase (SOD) and hepatic stellate cell receptor CB2than those in the model group increased significantly (P<0.01).Conclusion:Based on warm yang and expell cole’s hepatic ji powder downregulation of hepatic fibrosis rats serum ALT, AST, HA, LN, PC III, MDA, TGF beta1, liver tissue CTGF, TIMP-1, CTGFmRNA, TIMP-1mRNA, hepatic stellate cell receptor CB1, activation of hepatic stellate cells express; elevated serum SOD and liver stellate cell receptor CB2expression may be the part of the mechanism of anti hepatic fibrosis.更多还原