冠心病急性心肌梗死气阴两虚证动物模型的构建与评价  

Establishment and Evaluation of Animal Model of Acute Myocardial Infarction with Syndrome of Qi and Yin Deficiency

作　　者：高云霄 张秋艳 彭菊琴 郭浩[1] 陈潇潇 郝伟[1] 胡珑潇 史亚丽 任钧国[1] 刘建勋[1] GAO Yunxiao;ZHANG Qiuyan;PENG Juqin;GUO Hao;CHEN Xiaoxiao;HAO Wei;HU Longxiao;SHI Yali;REN Junguo;LIU Jianxun(National Clinical Research Center of Traditional Chinese Medicine for Cardiovascular Diseases,Beijing Key Laboratory of Chinese Materia Pharmacology,Institute of Basic Medical Sciences,Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China;Xiyuan Clinical Medical College,Beijing University of Chinese Medicine,Beijing 100029,China)

机构地区：[1]中国中医科学院西苑医院基础医学研究所,中药药理北京市重点实验室,国家中医心血管疾病临床医学研究中心,北京100091 [2]北京中医药大学临床医学院,北京100029

出　　处：《中国实验方剂学杂志》2024年第4期134-142,共9页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家科技重大专项重大新药创制(2017ZX09301018);中国中医科学院科技创新工程重大攻关项目(CI2021A04601)。

摘　　要：目的:探索睡眠剥夺(SD)复合异丙肾上腺素(ISO)建立冠心病急性心肌梗死气阴两虚证大鼠模型的构建和评价方法。方法:将40只SD大鼠分为正常组、SD组、ISO组、SD+ISO组。正常组不予处理,SD组使用改良多平台水环境法连续睡眠剥夺96 h,ISO组皮下注射ISO 100 mg·kg^(-1),隔日1次,共2次。SD+ISO组在连续睡眠剥夺72、96 h时注射ISO 100 mg·kg^(-1)。采用小动物超声心动图检测大鼠心功能;生化分析法检测血清肌酸激酶(CK)、肌酸激酶同工酶(CK-MB)、乳酸脱氢酶(LDH)、心肌肌钙蛋白T(cTnT)水平;苏木素-伊红(HE)染色观察心肌组织病理变化;通过一般状态及体质量、抓力、体温、旷场实验、血清环磷酸腺苷(cAMP)、环磷酸鸟苷(cGMP)及cAMP/cGMP、舌象红(R)、绿(G)、蓝(B)值及脉搏幅度观察气阴两虚证的证候指标变化;酶联免疫吸附测定法(EILSA)检测大鼠血清白细胞介素-18(IL-18)、肿瘤坏死因子-α(TNF-α)、氧化应激超氧化物歧化酶(SOD)、丙二醛(MDA)、促肾上腺皮质激素释放因子(CRF)、促肾上腺皮质激素(ACTH)、三碘甲状腺原氨酸(T3)、四碘甲状腺原氨酸(T4)及白细胞分化抗原4(CD4)、白细胞分化抗原8(CD8)水平。结果:疾病指标方面,与正常组比较,ISO组和SD+ISO组心功能降低(P<0.01),各模型组心肌酶升高(P<0.01),ISO组和SD+ISO组心肌组织病理损伤明显。证候指标方面,与正常组比较,SD组和SD+ISO组的各时间点体质量下降(P<0.01)、ISO组48、72 h体质量下降(P<0.05,P<0.01),各模型组抓力减弱,舌面R、G、B值降低(P<0.01),SD组爪温、肛温升高(P<0.01),各模型组不动时间延长(P<0.01),总路程和进入中心区次数下降(P<0.01),平均速度减缓(P<0.05,P<0.01);cAMP、cGMP升高(P<0.05,P<0.01),SD+ISO组cAMP/cGMP增加(P<0.01),SD组和SD+ISO组脉搏幅度降低(P<0.01)。血清学指标方面,与正常组比较,ISO组和SD+ISO组IL-18、TNF-α、SOD、MDA升高(P<0.01);各模型组的CRF、ACTH、CORT、T3、Objective:To explore the establishment and evaluation methods of the rat model of acute myocardial infarction(AMI)in coronary heart disease with the syndrome of Qi and Yin deficiency by sleep deprivation(SD)combined with isoproterenol(ISO)and preliminarily explore its biological basis.Method:Forty SD rats were assigned into normal(no treatment),SD(treatment in modified multi-platform water environment for 96 h),ISO(subcutaneous injection of ISO at 100 mg·kg^(-1) once every other day for a total of 2 times),and SD+ISO(injection of 100 mg·kg^(-1) ISO after SD for 72 h and 96 h)groups.The cardiac function was detected by small animal echocardiography.The serum levels of creatine kinase(CK),creatine kinase isoenzyme(CK-MB),lactate dehydrogenase(LDH),and cardiac troponin T(cTnT)were measured by biochemical methods.The pathological changes of the myocardial tissue were observed by hematoxylin-eosin staining.The general state,body weight,grip strength,body temperature,behaviors in open field test,serum levels of cyclic adenosine monophosphate(cAMP),cyclic guanosine monophosphate(cGMP),cAMP/cGMP ratio,red(R),green(G),blue(B)values of the tongue surface,and pulse amplitude were observed and measured to evaluate the modeling results.Enzyme-linked immunosorbent assay was employed to determine the serum levels of interleukin-18(IL-18),tumor necrosis factor-α(TNF-α),superoxide dismutase(SOD),malondialdehyde(MDA),corticotropin-releasing factor(CRF),adrenocorticotropic hormone(ACTH),triiodothyronine(T3),tetraiodothyronine(T4),cluster of differentiation 4(CD4),and cluster of differentiation 8(CD8).Result:In terms of disease indicators,the ISO and SD+ISO groups had lower cardiac function indicators than the normal group(P<0.01).The levels of CK,CM-MB,LDH and cTnT elevated in each model group compared with the normal group(P<0.01).The pathological changes of myocardial tissue were obvious in the ISO and SD+ISO groups.In terms of syndrome indicators,compared with the normal group,the SD and SD+ISO groups showed decreased body w

关 键 词：动物模型 急性心肌梗死 气阴两虚证 病证结合 

分 类 号：R2-0[医药卫生—中医学] R33R289R541R825.4