左心室重构大鼠模型的中医证候学评价及其相关中药的电生理作用研究

【作者】 倪量；

【导师】 王硕仁；

【作者基本信息】 北京中医药大学 ， 中西医结合临床内科学， 2007， 博士

【摘要】 心律失常是严重威胁人类健康的心脏疾病,严重的心律失常已经成为导致人类死亡的主要原因。而长期以来,有效治疗心律失常是中医药和中西医结合临床工作的薄弱环节。因此本研究致力于开展中西医结合防治心律失常的研究。目的:1.建立部分缩窄大鼠腹主动脉致心室重构动物模型,并对该模型的中医证候学进行研究;2.研究心室重构大鼠模型的离体心脏电生理特点,并以应用活血、益气中药的提取物为主进行防治心室重构致心律失常的药理研究。方法:1.用部分缩窄大鼠腹主动脉的方法,复制大鼠心室重构的动物模型。应用超声心动技术和病理实验技术(对模型大鼠心脏常规石蜡切片,分别行HE染色和Masson染色,显微图象观察分析)的方法,对造模后第4周和第8周的大鼠心脏进行病理学评价。以中西医结合学会的虚证、血瘀证辨证标准作为动物模型的辨证依据,采集动物模型的外在表观信息和心功能评价等内容、将临床定性的问诊内容替代以同等意义的实验测试指标,包括左心室射血分值、心电图ST段改变、力竭性游泳时间等,开展病证动物模型的综合评价。2.用结扎大鼠冠状动脉致心肌梗死的方法和按方法1,复制两种心室重构的大鼠动物模型。应用Langdorff灌注大鼠离体心脏的电生理实验的方法分别观察造模后第8周两种心室重构大鼠离体心脏的电生理指标,包括:动作电位复极20%的时间(APD20,ms),动作电位复极50%的时间(APD50,ms),动作电位复极90%的时间(APD90,ms),ERP / APD90比值,有效不应期(ERP,ms) ,动作电位最大幅度(APA,mv),零相上升的最大速率(Vmax,v/s),APD90离散度,ERP离散度。研究它们各自的电生理特点,比较它们之间电生理特点的异同。3.按上述灌注离体心脏的电生理实验方法,以经典的抗心律失常西药利多卡因、胺碘酮、维拉帕米为阳性对照药,研究益气、活血、清热中药的提取物,人参茎叶皂苷、川芎嗪、人参茎叶皂苷+川芎嗪、槐定碱、小檗碱对部分缩窄大鼠腹主动脉致心室重构动物模型的离体心脏电生理的影响。4.按上述灌注离体心脏的电生理实验方法,研究扶正化瘀胶囊(0.4g/kg体重)和卡托普利(2.2mg/kg体重)对结扎大鼠冠状动脉模型灌胃8周后的离体心脏电生理作用。结果:1.造模后第4周,部分缩窄大鼠腹主动脉致心室重构大鼠心脏,室间隔舒张末期厚度(IVSTd)增厚、左室重量指数(LVMI)增大、左室壁厚度增加、左室腔面积增大、心肌胶原容积分数(CVF)、心脏血管周围胶原面积比值(PVCA)增大、ST段发生缺血性变化导联增加,以上超声、病理、心电图指标与假手术组比较均有非常显著性差异(P <0.01),心率增快(P <0.05);力竭游泳时间缩短(P <0.01)。造模后第8周,IVSTd仍非常增加、左室后壁舒张末期厚度(LVPWTd)增加,左室壁厚度增大,心肌CVF、心脏PVCA增大,ST段发生缺血性变化导联增加,以上超声、病理、心电图指标与假手术组比较均有非常显著性差异(P <0.01),左室舒张末期内径(LVDd)增加( P <0.05),左室射血分数(LVEF)减小(P <0.05);LVMI增大(P<0.01),左室腔面积增大(P <0.05),心率增快(P <0.05);力竭性游泳时间缩短(P <0.01),以上数值与8周假手术组比较,均有显著性差异。2.造模后第8周,部分缩窄大鼠腹主动脉致心室重构大鼠离体心脏左心室动作电位复极20%(APD20)、动作电位复极50%(APD50)、动作电位复极90%(APD90)均非常显著延长(P <0.01)、有效不应期(ERP)有显著延长(P <0.05)、ERP/ APD90比值显著减小(P <0.05),右心室APD20、APD50、APD90、ERP、ERP/ APD90比值无明显变化,APD90离散度、ERP离散度非常显著增大(P <0.01)。结扎大鼠冠状动脉致心室重构大鼠离体心脏左心室APD20、APD50、APD90、ERP均非常显著延长、ERP/ APD90比值非常显著减小,右心室APD20、APD50、APD90、ERP均非常显著延长, ERP / APD90比值非常显著减小,APD90离散度、ERP离散度非常显著增大。以上各指标均与假手术组相应各指标比较有非常显著性差异(P <0.01)。3.各药对部分缩窄大鼠腹主动脉致心室重构大鼠离体心脏的电生理影响:(1)利多卡因:缩短心肌APD90时程、增加ERP/APD90比值,减小心肌动作电位复极离散度,减小APA、Vmax。(2)胺碘酮:延长心肌APD90、ERP时程、增加ERP/APD90比值;减小心肌动作电位和有效不应期的离散度,延长A-H时长,减慢房室结传导速度。(3)维拉帕米:延长心肌A-H时长,对于该模型的动作电位振幅(APA)、动作电位上升最大速率(Vmax)、时程、均无影响,对室性心律失常作用有限。(4)人参茎叶皂苷:A-H延长(P<0.01)。(5)川芎嗪:ERP延长(P<0.05),ERP / APD90比值增大(P<0.05),A-H延长(P<0.01)。(6)人参茎叶皂苷+川芎嗪:APD90、ERP延长(P<0.05),ERP / APD90比值增大(P<0.05),A-H延长(P<0.01)。(7)槐定碱:APD90、ERP延长(P<0.05),ERP / APD90比值增大(P<0.05),A-H、H-V间期延长(P<0.01)。(8)小檗碱APD90、ERP延长(P<0.05),ERP / APD90比值增加(P<0.05),APA和Vmax减小(P<0.01),A-H、H-V间期均延长(P<0.01)。4.卡托普利和扶正化瘀胶囊对结扎大鼠冠状动脉致心室重构模型的离体心脏电生理的影响:(1)卡托普利:原已增大的心肌APD90减小(P <0.05),原已减小的ERP / APD90比值增大(P <0.05),APD90离散度减小,致室颤的乌头碱用药量增大(P <0.05)。(2)扶正化瘀胶囊:原已增大的APD90减小(P <0.05),原已减小的ERP / APD90比值增大(P <0.05),致室颤的乌头碱用药量增大(P <0.05)。结论:1.部分缩窄大鼠腹主动脉致心室重构模型大鼠心脏,具有早期呈现典型的向心性肥厚的病理改变,以后发展为向心性肥厚又有离心性肥厚的混合型左室肥厚的动态改变。中医证候表现为气虚、血瘀证候的特点。2.两种心室重构大鼠离体心脏的电生理特性各有其不同的特点。部分缩窄腹主动脉致心室重构大鼠离体心脏左心室APD、ERP延长,右心室APD、ERP无改变。心梗后心室重构大鼠离体心脏左心室APD、ERP延长更加明显,右心室APD和ERP也明显延长。3.人参茎叶皂苷加川芎嗪合用后对心肌电生理的影响比单用人参茎叶皂苷和川芎嗪全面,具体是:既延长心肌APD90、ERP,又增加了ERP/APD90比值,同时也具有延长A-H时长;增加冠脉流量的作用。对于改善心肌APD90、ERP、ERP/APD90比值,增加冠脉流量的强度大于单独应用人参茎叶皂苷或川芎嗪。槐定碱能够显著延长心肌APD90、ERP,增加ERP/APD90比值,延长A-H时长,H-V时长等抗心律失常作用,但有显著减少了心室重构大鼠离体心脏冠脉流量的作用。小檗碱延长心肌APD90、ERP,增加ERP/APD90比值,延长了A-H时长和H-V时长等抗心律失常作用。但有减小APA和Vmax的负性电生理作用。4.对未曾报告有抗心律失常的药物扶正化瘀胶囊和卡托普利,经8周灌胃给药,显著改善心梗大鼠离体心脏的左右心室电生理指标。并增大了引发室颤的乌头碱用药量。更多还原

【Abstract】 Arrhythmia,which is a serious threat to human health, is a kind of heart disease. Serious arrhythmia has become the leading cause of death. And the long-standing, effective treatment of arrhythmia is the weak links for Chinese medicine and combination of TCM with Western medicine. Therefore this study is to launch prevention and treatment of Arrhythmia Research.Objectives:1. The establishment of coarctation rats abdominal aorta induced animal model of ventricular remodeling, and the research of traditional Chinese medicine syndrome.2. To study electrophysiological characteristics of isolated heart of ventricular remodeling in rats, and to make pharmacological research of application of active Blood and Yiqi medicine extracts to treat arrhythmia, which caused by ventricular remodeling.Methods:1. To duplicate the rat ventricular remodeling in animal models with coarctation rats abdominal aorta. Application of echocardiography technology and pathology laboratory to evaluate the pathologic characteristics of rat hearts right model after four weeks and eight weeks. Based on diagnostic standard of Integrated Traditional Chinese and Western Medicine about deficient syndrome and syndrome of blood stasis, with acquisition of the apparent external information and evaluation of cardiac function, conduct animal disease and syndrome comprehensive evaluation in animal model.2. Application of coronary artery ligation and abdominal aorta coarctation in rats to duplicate two kinds of ventricular remodeling animal model. With the method of Langdorff reperfusion isolated rat cardiac electrophysiology experimental, electrophysiology index, which including 20% action potential duration(APD20,ms), 50% action potential duration(APD50,ms), 90% action potential duration(APD90,ms), effective refractory period(ERP,ms), action potential amplitude(APA,mv), maximum velocity of phase depolarization(Vmax,v/s), APD90 divergence and ERP divergence, at 8 weeks right after operation. To study its electrophysiological characteristics. 3. With the above in vitro perfusion of cardiac electrophysiology experimental, using lidocaine, amiodarone, verapamil as a positive control, make the research about electrophysiology impact of pharmaceutics remedies (gensenosides, trtram-ethylpvrazine, gensenosides and trtram-ethylpvrazine, sophordine, berbeine) on the rat ventricular remodeling in animal models with coarctation rats abdominal aorta.4.With the above in vitro perfusion of cardiac electrophysiology experimental, studying the electrophysiology impact of Fuzhenghuayu caps(0.4g/kg)and captopril(2.2mg/kg) on the rat ventricular remodeling in animal models with coronary artery ligation.Results:1. At 4 weeks of operation, in rat ventricular remodeling in animal models with coarctation rats abdominal aorta, inter-ventricular septum end-diastolic thickness(IVSTd),left ventricular mass index(LVMI),left ventricular wall thickness, left ventricular cavity area, collagen volume fraction (CVF), Perivascular Collagen area (PVCA) and leads of ST-segment changes increased (P <0.01), heart rate increased (P <0.05), exhaustive swim time decreased (P <0.01). At 8 weeks of operation, in rat ventricular remodeling in animal models with coarctation rats abdominal aorta, IVSTd, left ventricular posterior wall end-diastolic thickness(LVPWTd), left ventricular wall thickness, left ventricular cavity area, CVF, PVCA and leads of ST-segment changes increased (P <0.01), left ventricular end-diastolic diameter(LVDd) increased ( P <0.05 ) , left ventricular ejection fraction(LVEF) decreased(P <0.05), LVMI increased(P<0.01), left ventricular cavity area increased(P <0.05), heart rate increased (P <0.05), exhaustive swim time decreased (P <0.01).2. At 4 weeks of operation, in rat ventricular remodeling in animal models with coarctation rats abdominal aorta, electrophysiology index of left ventricle, including APD20,APD50 APD90 increased ( P <0.01 ) , ERP increased ( P <0.05),ERP/ APD90 decreased(P <0.05).But electrophysiology index of right ventricle have no changes. In rat ventricular remodeling in animal models with coronary artery ligation, electrophysiology index of left and right ventricle all increased.3.Electrophysiology impacts of drugs on the isolated heart of animal models with coarctation rats abdominal aorta. (1) Lidocaine : shorten the time of APD90 , increased the ratio of ERP/APD90, reduced myocardial action potential repolarization dispersion, reduced the APA, Vmax. (2) Amiodarone: extended the time of APD90 and ERP, increased the ratio of ERP/APD90, reduced myocardial action potential and effective refractory period of dispersion, extended the time of A-H and slow down the AV node conduction velocity. (3) Verapamil: extended the time of A-H. (4) GSL: extended the time of APD90, ERP (P <0.05) and A-H (P <0.01). (5) Trtramethylpyrazine (TMP): extended the time of ERP (P <0.05) and A-H (P <0.01), increased the ratio of ERP/APD90 (P <0.05). (6) GSL + TMP: extended the time of APD90 and ERP (P <0.05), increased the ratio of ERP/APD90 (P <0.05), extended the time of A-H (P <0.01). (7) Sophoridine: extended the time of APD90 and ERP (P <0.05), increased the ratio of ERP/APD90 (P <0.05), extended the time of A-H and H-V (P <0.01). (8) Berberine: extended the time of APD90 and ERP (P <0.05) increased the ratio of ERP / APD90 (P <0.05), decreased APA and Vmax (P <0.01), extended the time of A-H (P <0.01).4.Electrophysiology impacts of captopril and Fuzhenghuayufang capsule on the isolated heart of animal models with coronary artery ligation: (1) captopril: Shorten the time of APD (P <0.05), increased the ratio of ERP/APD90 (P <0.05), decreased the APD90 dispersion, induced ventricular fibrillation in aconitine dosage increased (P <0.05). (2) Fuzhenghuayufang Capsule: Shorten the time of APD (P <0.05), increased the ratio of ERP/APD90 (P <0.05), decreased the APD90 dispersion, induced ventricular fibrillation in aconitine dosage increased (P <0.05).Conclusions:1.The heart of animal models with coarctation rats abdominal aorta present typical concentric hypertrophy in early, and development combination of concentric hypertrophy and eccentric hypertrophy after sometimes. The characteristic of syndrome of traditional Chinese medicine present syndrome of Qixu and blood stasis.2. The isolated heart of two kinds of ventricles remodeling have their own electrophysiological characteristics.3. Combination gensenosides with trtram-ethylpvrazine have better electrophysiological impact on the heart of animal models with coarctation rats abdominal aorta than gensenosides or trtram-ethylpvrazine.4. Fuzhenghuayu caps and captopril, which have never been report to have antiarrhythmic function, had been found that their have some good electrophysiological impact on the animal models with coronary artery ligation.更多还原