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降低(P<0.05,P<0.01),模型组肝组织形态学检查发现有假小叶形成,且汇管区出现典型的病变,阳性药组上述病变可呈现不同程度改善。结论:改良后的大鼠肝硬化肝郁脾虚证模型建立方法可提高造模成功率,降低造模导致大鼠的死亡率,具有成模时间短,与临床肝硬化肝郁脾虚证的证候吻合度高、可行性好,证候表现稳定的特点,以适宜于肝硬化肝郁脾虚证临床方证相关基础性研究和中药复方活性发现、药效评价与机制探讨。 OBJECTIVE: To construct a stable and reliable model of liver-qi stagnation and spleen-deficiency syndrome combining with the clinical manifestations of liver-qi stagnation syndrome and spleen-deficiency syndrome of liver cirrhosis. Methods: Seventy rats were randomly divided into normal group according to body weight. In model group Ⅰ and silybin group Ⅰ, the rats were treated with 79.5% pure corn meal plus 20% lard and 0.5% cholesterol. Protein feed; 30% ethanol composite factors for 4 weeks; improved model group Ⅱ, silybin group meglumine Ⅱ: 79.5% pure corn meal plus 20% lard, 0.5% cholesterol High-fat and low-protein feed, 10% ethanol for 1 week and 5% ethanol for 3 weeks. The rats in model group Ⅲ and silybin-meglumine Ⅲ were treated with 89.5% pure corn flour Plus 10% lard, 0.5% cholesterol made of high fat and low protein feed, ethanol gavage method with the model group Ⅱ; silybin megakaryon tablets Ⅰ, Ⅱ, Ⅲ were 0.108 g · kg-1 Silybin meglumine tablets gavage treatment. During the experimental period, the rest rats in each group were given the same degree of unpredictable emotional stimuli except the normal group. The general state of rats in each group was observed daily to simulate clinical manifestations of liver-spleen deficiency syndrome of liver cirrhosis. The syndromes of rats were observed. The body weight and the amount of water on solar eclipse were recorded daily. The serum levels of alanine aminotransferase (ALT), aspartate aminotransferase AST, HA, PC-Ⅲ, IV-C, ADA, MAO, A / G, Hardening closely related indicators of the level of hematoxylin-eosin (HE) staining observed pathological changes in each group. Results: In the normal group, there was no death of the rats and the nutritional status was good. In the model group, the diets of drinking water decreased, the body weight increased slowly, the hair became rough and the spirit was sluggish. 3 died, and there was no death in model group Ⅲ. Compared with normal group, serum ALT, AST, HA, PC-Ⅲ, IV-C, ADA and MAO in model group were significantly increased, A / G was significantly decreased, <0.01). The serum levels of ALT, AST, HA, PC-Ⅲ, IV-C, ADA and MAO of rats in each administration group were significantly lower than those in model group Ⅰ, A / G was significantly increased, (P <0.05, P <0.01). The formation of pseudolobule was observed in the liver tissue of the model group, and the typical lesions appeared in the portal area. The lesions of the positive drug group showed different degrees of improvement. Conclusion: The improved model of liver-qi deficiency and spleen-deficiency syndrome in rats with cirrhosis of liver can improve the successful rate of model making and reduce the death rate of rats caused by model making. It has the advantages of short formation time and syndrome of liver-qi deficiency and spleen-deficiency syndrome in clinical cirrhosis High coincidence degree, good feasibility and stable syndrome. It is suitable for the basic research on the clinical prescription of liver-qi stagnation and spleen-deficiency syndrome and the activity discovery, the evaluation of the efficacy and the mechanism of traditional Chinese medicine.