冠心病血瘀证形成过程中心肌细胞能量代谢酶作用及其机制研究  

Effect and Mechanism of Energy Metabolism Enzymes in Cardiomyocytes during the Formation of Coronary Heart Disease with Blood Stasis Syndrome

作　　者：周曼丽 俞赟丰 冯宇 简维雄 ZHOU Manli;YU Yunfeng;FENG Yu;JIAN Weixiong(College of Traditional Chinese Medicine,Hunan University of Chinese Medicine,Changsha 410208,China;National Key Discipline of TCM Diagnositics/Hunan Provincial Key Laboratory,Hunan University of Chinese Medicine,Changsha 410208,China)

机构地区：[1]湖南中医药大学中医学院,湖南省长沙市410208 [2]湖南中医药大学国家重点学科中医诊断学实验室湖南省重点实验室,湖南省长沙市410208

出　　处：《实用心脑肺血管病杂志》2020年第12期76-84,共9页Practical Journal of Cardiac Cerebral Pneumal and Vascular Disease

基　　金：国家自然科学基金资助项目(81774207,81973753);湖南省自然科学基金资助项目(2018JJ2291)。

摘　　要：背景冠心病血瘀证是一个“血瘀证前期”→“亚血瘀证期”→“心血瘀阻证期”流动的过程,心肌细胞能量代谢贯穿始终,而以往对能量代谢的研究局限于病程中的某个阶段,且在缺乏对冠心病血瘀证起始、转归整个过程进行研究时,这种孤立存在、缺乏联系的病理学现象并不能诠释心肌能量代谢的特点。因此,探究冠心病血瘀证形成过程中能量代谢改变尤为重要。目的分析冠心病血瘀证形成过程中心肌细胞能量代谢酶作用及其机制。方法本实验时间为2019年10—12月。采用简单随机抽样法将30只健康雄性SD大鼠分为空白对照组6只(A组)和模型大鼠24只。A组给予普通饲料喂养,模型大鼠高脂饲料喂养7 d后进行维生素D3灌胃(30万U/kg),4 d后再予以维生素D3灌胃(20万U/kg),继续高脂饲料喂养21 d后,死亡4只大鼠,从剩余20只大鼠中随机选择6只为血瘀证前期组(B组);剩余14只大鼠继续高脂饲料喂养30 d后死亡2只,从剩余12只大鼠中随机选择6只为亚血瘀证期组(C组);余6只大鼠继续高脂饲料喂养的同时予以皮下多点注射异丙肾上腺素(5 mg/kg),连续注射3 d,1周后记录标准Ⅱ导联心电图,以ST段出现上抬或明显压低(≥0.1 mV)确定成模,为血瘀证期组(D组)。观察并记录各组大鼠一般情况、血脂指标〔包括总胆固醇(TC)、三酰甘油(TG)、低密度脂蛋白胆固醇(LDL-C)、高密度脂蛋白胆固醇(HDL-C)水平〕、腹主动脉及心肌组织HE染色结果、心电图、线粒体超微结构、心肌组织能量代谢酶〔三磷酸腺苷(ATP)、一磷酸腺苷(AMP)、Na^+-K^+-ATP酶、Ca^2+-Mg^2+-ATP酶、酯酰辅酶A合成酶(ACS)、肉毒碱脂酰转移酶(CACT)〕水平。结果B组大鼠体质量减轻,反应渐迟钝,精神倦怠,毛色枯槁无光泽;C组大鼠体质量明显减轻,精神萎靡,蜷缩相拥,活动量少;D组大鼠精神欠佳,易惊,触之狂躁,毛色枯槁无光泽,爪甲紫暗。B组大鼠TC、LDBackground Coronary heart disease with blood stasis syndrome is a flow process of"pre-blood stasis syndrome"→"sub-blood stasis syndrome"→"heart blood stasis syndrome".Energy metabolism of cardiomyocytes runs through the whole process.Previous studies on energy metabolism were limited to a certain stage in the course of disease.In the absence of research on the whole process of initiation and outcome,this isolated pathological phenomenon with lack of connection cannot interpret the characteristics of myocardial energy metabolism.Therefore,it is particularly important to explore changes in energy metabolism during the formation of blood stasis syndrome in coronary heart disease.Objective To explore the effect and mechanism of energy metabolism enzymes in cardiomyocytes during the formation of coronary heart disease with blood stasis syndrome.Methods The study time was from October to December 2019.Thirty healthy male SD rats were randomly divided into blank control group(6 rats in group A)and 24 model rats.The rats in group A were fed with normal diet,and the model rats were fed with high-fat diet for 7 days,and then fed with vitamin D3(300000 U/kg),4 days later fed with vitamin D3(200000 U/kg).After 21 days of high-fat diet feeding,4 rats died and 6 rats were randomly selected as the blood stasis syndrome group from the remaining 20 rats(group B);the remaining 14 rats were fed with high-fat diet for 30 days,2 rats died and 6 rats were randomly selected as sub blood stasis syndrome group from the remaining 12 rats(group C);the remaining 6 rats were continuously fed with high-fat diet and subcutaneously injected with isoproterenol(5 mg/kg)for 3 consecutive days.One week later,ECG of standard leadⅡwas recorded,and the model was established by ST segment elevation or obvious depression(≥0.1 mV).And they were as the blood stasis syndrome group(group D).The general condition,blood lipid indexes(including TC,TG,LDL-C,HDL-C levels),HE staining results of abdominal aorta and myocardial tissue,ECG,mitochondrial ultr

关 键 词：冠心病 血瘀证 肌细胞 心脏 能量代谢 腺苷三磷酸 腺苷一磷酸 NA^+-K^+-ATP酶 CA^2+-MG^2+-ATP酶 酯酰辅酶A合成酶 肉毒碱脂酰转移酶 

分 类 号：R259[医药卫生—中西医结合]