瘀毒互结证动物模型的建立与评价  

Establishment and Evaluation of Animal Models of Combined Stasis and Toxin Syndrome

作　　者：刘悦 姚明江[1,2,3] 梁晓 崔文强 申伟 魏竞竞 迟显苏 刘红喜 刘建勋 张允岭[1] LIU Yue;YAO Mingjiang;LIANG Xiao;CUI Wenqiang;SHEN Wei;WEI Jingjing;CHI Xiansu;LIU Hongxi;LIU Jianxun;ZHANG Yunling(Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China;Institute of Basic Medical Sciences of Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China;Beijing Key Laboratory of Pharmacology of Chinese Materia Medica,Beijing 100091,China)

机构地区：[1]中国中医科学院西苑医院,北京100091 [2]中国中医科学院西苑医院基础医学研究所,北京100091 [3]北京市中药药理重点实验室,北京100091

出　　处：《中国实验方剂学杂志》2023年第13期72-78,共7页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家中医药管理局中医药创新团队及人才支持计划项目(ZYYCXTD-C-202007);中国中医科学院科技创新工程项目(CI2021A01301);中医药防治脑病传承创新团队项目(CI2021B006)。

摘　　要：目的:通过比较采用角叉菜胶(Ca)、角叉菜胶联合干酵母菌(Ca+Yeast)、角叉菜胶联合脂多糖(Ca+LPS)3种不同方式,筛选并建立瘀毒互结证动物模型。方法:将40只SPF级雄性SD大鼠随机分成4组,每组10只,包括正常组、Ca组、Ca+Yeast组、Ca+LPS组。Ca组、Ca+Yeast组、Ca+LPS组于造模首日经腹腔注射Ca 10 mg·kg^(-1)。Ca+LPS组于第2天经腹腔注射LPS 50μg·kg^(-1),Ca+Yeast组于第2天背部皮下注射干酵母悬液2 mg·kg^(-1)。造模后动态观察各组大鼠肛温变化情况;在造模后24 h,观察各组大鼠中医证候宏观评价指标舌象、脉象、黑尾长度,采用PeriCam PSI血流灌注散斑成像仪检测大鼠尾端血流灌注量,利用全自动血流变测试仪检测各组大鼠全血黏度值及血浆黏度值,PL^(-1)2型血小板功能检测仪检测大鼠血小板聚集率,酶联免疫吸附测定法(ELISA)检测大鼠血浆中白细胞介素(IL)-6的情况,苏木素-伊红(HE)染色观察各组大鼠心肌组织、脑组织和肺组织。结果:与正常组比较,3组模型组均出现不同程度的黑尾(P<0.05,P<0.01),尾端血流灌注量明显降低(P<0.05,P<0.01),舌象R、G、B值明显下降(P<0.05,P<0.01),且血小板最大聚集率明显升高(P<0.05,P<0.01)。Ca+Yeast组及Ca+LPS组脉搏幅度较正常组明显下降(P<0.05,P<0.01)。此外,与正常组比较,Ca+Yeast组在造模后24 h内的平均肛温显著升高(P<0.01);全血黏度低、中、高切及血浆黏度均明显升高(P<0.05,P<0.01);血浆炎症因子IL-6表达水平明显上升(P<0.05)。病理形态学观察结果显示,Ca+Yeast组病理改变最为严重,可观察到小灶心肌纤维溶解断裂,炎性细胞浸润,纤维细胞增生;海马区局部神经元稀疏,神经元红色变性;小灶肺泡间质内淋巴细胞和中性粒细胞浸润。结论:利用角Ca+Yeast能够成功建立瘀毒互结证动物模型,以中医四诊信息、西医微观指标及组织病理形态学为系统的模型评价体系值得研究者思考与借�Objective:To screen and establish animal models of combined stasis and toxin syndrome based on the comparison of three modeling methods,i.e.,carrageenan(Ca),Ca combined with dried yeast(Ca+Yeast),and Ca combined with lipopolysaccharide(Ca+LPS).Method:Forty SPF male SD rats were randomly divided into normal group,Ca group,Ca+Yeast group,and Ca+LPS group,with 10 rats in each group.The Ca group,Ca+Yeast group,and Ca+LPS group received an intraperitoneal injection of Ca(10 mg·kg^(-1))on the first day.The Ca+LPS group received an intraperitoneal injection of LPS(50μg·kg^(-1))on the second day,and the Ca+Yeast group received a subcutaneous injection of dry yeast suspension(2 mg·kg^(-1))on the back on the second day.The rectal temperature of each group was dynamically observed after modeling.After 24 hours of modeling,the macroscopic evaluation indexes,including tongue manifestation,pulse,and black tail length in each group were observed.The PeriCam PSI imaging system was used to detect the blood flow perfusion of the rat tail.The automatic hemorheology analyzer was used to measure the whole blood viscosity and plasma viscosity of each group.The PL platelet function analyzer was used to detect the platelet aggregation rate of the rats.The enzyme-linked immunosorbent assay(ELISA)was used to detect the interleukin-6(IL-6)level in the rat plasma.The myocardial tissue,brain tissue,and lung tissue of each group of rats were observed by hematoxylin-eosin(HE)staining.Result:Compared with the normal group,all three model groups showed varying degrees of black tail(P<0.05,P<0.01),reduced blood flow perfusion at the tail end(P<0.05,P<0.01),decreased R,G,and B values of tongue manifestation(P<0.05,P<0.01),and increased maximum platelet aggregation rate(P<0.05,P<0.01).The pulse amplitudes of the Ca+Yeast group and the Ca+LPS group were lower than that of the normal group(P<0.05,P<0.01).In addition,the average rectal temperature of the Ca+Yeast group increased after 24 hours of modeling(P<0.01),and the low-,medium-,and high-shea

关 键 词：瘀毒互结证 证候模型 角叉菜胶 干酵母菌 模型评价 

分 类 号：R2-0[医药卫生—中医学] R22R285.5R289R33