改良高脂及乙醇加情绪刺激诱导大鼠肝硬化肝郁脾虚证模型的建立  

Establishment of Hepatic Cirrhosis Models with Syndrome of Liver Depression and Spleen Deficiency in Rats by Modified High Fat and Alcohol Stimulation and Emotional Stimuli

作　　者：石亮[1,2] 孙晓倩[2] 李晓宇[1] 孙蓉[1] 

机构地区：[1]山东省中医药研究院,济南250014 [2]山东中医药大学,济南250355

出　　处：《中国实验方剂学杂志》2017年第14期106-112,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金面上项目(81374059);山东省自主创新和成果转化项目(2014ZZCX02104);山东省自然基金重点项目(2011ZRB14333);泰山学者工程专项经费项目(Ns201511107);山东省重点研发计划(重大关键技术)项目(2016ZDJS07A21)

摘　　要：目的:构建能与肝硬化肝郁脾虚证临床证候表现密切相关的、并适宜于中药活性发现和药效评价的稳定可靠的大鼠肝硬化肝郁脾虚证病证结合模型。方法:70只大鼠按体重随机分为正常组;模型Ⅰ组、水飞蓟宾葡甲胺片Ⅰ组:采用79.5%单纯玉米粉加20%猪油,0.5%胆固醇制成的高脂低蛋白饲料饲养;30%乙醇复合因素造模连续4周;改良模型Ⅱ组、水飞蓟宾葡甲胺片Ⅱ组:采用由79.5%单纯玉米粉加20%猪油,0.5%胆固醇制成的高脂低蛋白饲料饲养,10%乙醇灌胃1周,5%乙醇灌胃3周的方法造模;改良模型Ⅲ组、水飞蓟宾葡甲胺片Ⅲ组:采用由89.5%单纯玉米粉加10%猪油,0.5%胆固醇制成的高脂低蛋白饲料饲养,乙醇灌胃方法同模型Ⅱ组;水飞蓟宾葡甲胺片Ⅰ,Ⅱ,Ⅲ组均按0.108 g·kg-1灌胃水飞蓟宾葡甲胺片治疗。实验期间除正常组外其余各组大鼠均给予相同程度、不可预知的情绪刺激,每日观察各组大鼠一般状态,模拟临床肝硬化肝郁脾虚证临床证候表现,分析并提出适宜于大鼠的证候表现,并每日按积分标准进行赋分,记录大鼠的体重、日食日水量,实验结束眼球取血检测大鼠血清丙氨酸转氨酶(ALT),天门冬氨酸转氨酶(AST),透明质酸(HA),Ⅲ型前胶原(PC-Ⅲ),IV型胶原(IV-C),腺苷脱氨酶(ADA),单胺氧化酶(MAO)和A/G等与肝硬化密切相关指标的水平,苏木素-伊红(HE)染色观察各组肝组织病理学变化。结果:正常组无大鼠死亡,营养状况良好;模型组大鼠饮食饮水量降低,体重增加缓慢,毛质粗糙,精神萎靡,个别老鼠趾甲易出血;其中模型Ⅰ组死亡4只,模型Ⅱ组死亡3只,模型Ⅲ组无死亡情况。与正常组比较,模型组大鼠血清ALT,AST,HA,PC-Ⅲ,IV-C,ADA,MAO水平显著升高,A/G显著降低,脏脑比例及证候积分显著升高(P<0.01);各给药组大鼠血清ALT,AST,HA,PC-Ⅲ,IV-C,ADA,MAO水平与模型Ⅰ组相比明显降低,A/G明显升高,脏脑比例及证候积分明显Objective： To establish stable and reliable rat models of hepatic cirrhosis with syndrome of liver depression and spleen deficiency, closely related to clinical disease and suitable for Chinese medicine active discovery and efficacy evaluation. Method： The 70 rats were randomly assigned to normal control group; model group I （79.5% simple corn flour plus 20% lard, high fat and low protein feed composed of 0.5% cholesterol; 30% alcohol compound factors for four weeks of modeling）, silybin meglumine tablets group I; modified model group II （79.5% simple corn flour plus 20% lard, high fat and low protein feed composed of 0.5% cholesterol; 10% alcohol gavage for 1 week, and 5% alcohol gavage for 3 weeks for modeling）, silybin meglumine tablets group II; modified model group Ⅲ （89.5% simple corn flour plus 10% lard, high fat and low protein feed composed of 0.5% cholesterol; 10% alcohol gavage for 1 week, and 5% alcohol gavage for 3 weeks for modeling） , and silybin meglumine slice group Ⅲ according to their body weight, n = 10 in each group. The rats in Silibin meglumine tablets group I, group II and group m received the treatment of silibin meglumine tablets at the dose of 0. 108 g.kg-1by gavage administration. During the experimental period, rats in all the other groups except the normal control group received unexpected emotional stimuli. Their general state was observed daily in every group ; the clinical syndrome of liver cirrhosis （ liver stagnation and spleen deficiency syndrome） was simulated ; the syndrome performance suitable for the rats were analyzed and put forward; and the scores were given according to the standard every day. In addition, the body weight of the rats, daily food intake and daily water intake were recorded. The contents of alanine aminotransferase （ALT）, aspartate aminotransferase （AST）, hyaluronic acid （HA） , procollagen Ⅲ（ PC Ⅲ ） , type collagenase IV （IV-C） , adenosine deaminase （ADA） , monoamine oxidase （MAO） and A/G in serum we

关 键 词：改良高脂 肝硬化 水飞蓟宾葡甲胺片 乙醇加情绪 肝郁脾虚证 

分 类 号：R285.5[医药卫生—中药学]