痰瘀互结证慢性心肌缺血小型猪模型的建立

目的 采用高脂、静脉注射维生素D₃(VD₃)和异丙肾上腺素建立痰瘀互结证慢性心肌缺血小型猪模型。方法 15只雄性巴马小型猪随机分成正常对照(Ctr)组、高脂(HFC)组和痰瘀互结证心肌缺血模型(CMI)组,每组5只,Ctr组饲喂普通饲料,HFC组饲喂高脂饲料,CMI组采用高脂饮食、静脉注射VD₃和异丙肾上腺素联合建立痰瘀证慢性心肌缺血模型,连续造模24周,分别通过测定动物体重、心电图、活动、血脂、心肌酶、血液流变学、炎症、心脏指数(CI)和心肌缺血面积(MIS)来评价模型的建立及痰瘀互结证的病理进程。结果 与Ctr组,HFC组体重、心率(HR)、总胆固醇(TC)、低密度脂蛋白(LDL-C)、高密度脂蛋白(HDL-C)、动脉粥样硬化指数(AI)、低、中、高切全血粘度和还原粘度、红细胞电泳时间(EPT)、超敏C-反应蛋白(hs-CRP)和白介素-6(IL-6)水平均明显升高(P<0.05, P<0.01);CMI组体重、HR、24 h总的ST段升高数(∑ST)、24 h内平均ST段升高数(ST_average)、活动度、TC、甘油三酯(TG)、LDL-C、HDL-C、AI、肌酸激酶(CK)、乳酸脱氢酶(LDH)、肌钙蛋白-1(cTn-1)、低、中、高切全血粘度和还原粘度、EPT、卡松粘度(CV)、hs-CRP、IL-6、CI和MIS均显著升高(P<0.05, P<0.01),脂联素(APN)水平则显著降低(P<0.05);与HFC组比,CMI组AI、CK、LDH、cTn-1、低、中、高切全血粘度、EPT、CI和MIS均明显高于HFC组(P<0.05, P<0.01),且APN水平则显著低于HFC组(P<0.05)。相关分析显示,MIS与TC、LDL-C、AI、CK、LDH、cTn-1、APN、高、中、低切全血粘度、EPT、CV、hs-CRP和IL-6有关(P<0.05, P<0.01),同时线性回归分析显示,痰瘀与TC、LDL-C、AI、CK、LDH、cTn-1、APN、EPT、CV、hs-CRP和IL-6有关(P<0.01),进一步线性逐步回归分析显示痰瘀衍变与TC、CK和IL-6密切相关(P<0.01)。结论 高脂喂养和注射VD₃和异丙肾上腺素能成功建立痰瘀互结证慢性心肌缺血小型猪模型,且脂质代谢、血液流变学、心肌酶代谢和炎症可作为痰瘀互结证的客观生化物质基础,这些变化可反映了中医理论"痰瘀互结、瘀毒致变"的相关生物学基础。

Establishment of chromic myocardial ischemia model of phlegm-blood stasis syndrome type in miniature swine

Objective To establish a disease syndrome combined animal model, the miniature pig model of chronic myocardial ischemia of phlegm-blood stasis syndrome type, by high fat/cholesterol diet feeding and intravenous injection with VD₃ and isoproterenol. Methods Miniature pigs were randomly divided into the control (Ctr) group, high fat/cholesterol diet (HFC) group and chronic myocardial ischemia model of phlegm-blood stasis syndrome (CMI) group, 5 pigs in each group. The Ctr group was fed with normal regular chow diet, HFC group was fed with high fat/cholesterol diet, while the CMI group was fed with high fat/cholesterol diet and intravenous injection with VD₃ and isoproterenol. The experiment lasted for 24 weeks. The model establishment and its pathological process of phlegm-blood stasis syndrome were evaluated by examinations of body weight, electrocardiogram, activity, blood lipid, myocardial enzymes, hemorheology, inflammation, cardiac index(CI) and myocardial ischemia size (MIS). Results Compared with the Ctr group, the body weight, heart rate(HR), Total cholesterol(TC), low density lipoprotein-cholesterol (LDL-C), high density lipoprotein-cholesterol(HDL-C),atherosclerosis index(AI),low/middle/high shear rate of whole blood viscosity and reduced viscosity, erythrocyte electrophoresis time(EPT), high-sensitivity C-reactive protein (hs-CRP) and IL-6 levels in the HFC group were significantly increased (P<0.05, P<0.01), while the body weight, heart rate, total ST, ST_average, activity, TC, TG, LDL-C, HDL-C, AI, CK, LDH, cTn-1, low/middle/high shear rate of whole blood viscosity and reduced viscosity, EPT, Casson viscosity(CV), hs-CRP, IL-6, CI and MIS in the CMI group were significantly increased (P<0.05, P<0.01), and APN level in the CMI group was significantly decreased (P<0.05). Moreover, AI, CK, LDH, cTn-1, low/middle/high shear rate of whole blood viscosity, EPT, CI and MIS in the CMI group were significantly higher than those of HFC group (P<0.05, P<0.01), while APN in the CMI group was significantly lower than that of HFC group (P<0.05). Correlation analysis showed that MIS was closely correlated to TC, LDL-C, AI, CK, LDH, cTn-1, APN, high/middle/low shear rate of whole blood viscosity, EPT, CV, hs-CRP and IL-6 (P<0.05, P<0.01). The linear regression analysis also showed that phlegm-blood stasis was closely correlated to TC, LDL-C, AI, CK, LDH, cTn-1, APN, CV, EPT, hs-CRP, and IL-6 (P<0.01), and further linear stepwise regression analysis showed that the evolution of phlegm-blood stasis was closely related to TC, CK and IL-6. Conclusions Minipig model of chronic myocardial ischemia of phlegm-blood stasis syndrome type can be established by high fat/cholesterol diet feeding and intravenous injection with VD₃ and isoproterenol. Their blood lipid metabolism, hemorheology, myocardial enzymes and inflammatory indexes can be used as external biochemical basis of phlegm-blood stasis syndrome type, which may reflect related biological basis of the traditional Chinese medicine theory of "phlegm and stasis cementation, blood-stasis & toxin causing catastrophe".