非酒精性脂肪性肝病痰瘀互结证大鼠模型的构建及评价

Construction and Evaluation of Rat Model of Nonalcoholic Fatty Liver Disease with Intermingled Phlegm and Blood Stasis Syndrome

目的探讨非酒精性脂肪性肝病(NAFLD)痰瘀互结证大鼠模型的构建。方法将20只SD大鼠随机分成对照组、模型组、经方组、协定方组,每组5只。除对照组外其余各组采用伤湿法加外界慢性轻度刺激、高脂饮食多因素复合刺激的方法建立NAFLD痰瘀互结证大鼠模型,连续10周。经方组和协定方治组于造模8周后给予相应中药1 ml/100 g灌胃,对照组和模型组均使用相同体积的生理盐水灌胃,每日1次,共2周。各组分别于造模1、10周进行舌象评分。造模10周进行肝脏HE染色,观察肝细胞脂肪变性程度;检测血脂、肝脏酶学指标及血液流变学指标。结果与对照组比较,模型组动物舌象评分、肝脏酶学指标、血脂、血液流变学指标均升高(P<0.05)。经方组和协定方组舌下络脉评分较同时间模型组降低,血脂及肝脏酶学指标水平亦降低(P<0.05);协定方组胆固醇(TC)、甘油三酯(TG)、丙氨酸氨基转移酶(ALT)水平较经方组下降明显(P<0.05)。结论采用伤湿法加外界慢性轻度刺激、高脂饮食多因素复合刺激的方法建立的大鼠模型病理符合NAFLD,同时具有痰瘀互结证中医证候,中药方剂干预后反证造模成功。

Objective To explore the modeling method of rat model of nonalcoholic fatty liver disease（ NAFLD）with intermingled phlegm and blood stasis syndrome. Methods Twenty Sprague Dawley rats were randomized into a control group,a model group,a classical prescription group,and a proved prescription group,n = 5 in each group.Rats in all groups except those in the control group were exposed to the damp environment,multiple factors compound stimulation,external chronic mild irritation,and high fat diet for 10 weeks to establish the models of NAFLD with intermingled phlegm and blood stasis syndrome. After 8 weeks of modeling,the classical prescription group and the proved prescription group were given 1 ml/100 g of corresponding herbal medicine by gavage. The control group and the model group were given equivalent normal saline by gavage,once every day,for 2 weeks. At the 1 st and 10 th week of modeling,tongue manifestation scores（ including tongue color and sublingual vessel） in each group were recorded. At the 10 th week of modeling,liver hematoxylin and eosin（ HE） stain was taken to examine the level of hepatic steatosis. Blood fat and liver enzymology indexes as well as hemorheology indexes including plasma viscosity and whole blood viscosity were detected. Results Compared with those in the control group,rats＇ tongue manifestation scores,liver enzymology indexes,blood fat and hemorheology indexes increased in the model group（ P 0. 05）. The sublingual vessel score in the classical prescription group and the proved prescription group were lower,and the levels of blood fat and liver enzymology indexes decreased compared with that in the model group at the same time（ P 0. 05）. The levels of total cholesterol（ TC）,triglyceride（ TG） and alanine aminotransferase（ ALT） in the proved prescription group were lower than those in the prescription group（ P 0. 05）. Conclusion The pathology of rat model induced by damp environment,multiple factors compound stimulation,external chronic mild irritation and high fat diet is in accordance with NAFLD,and the model is with Chinese medicine syndrome of intermingled phlegm and blood stasis. After herb-prescription intervention,it is proved by contradiction that model construction is successful.