高脂饲料联合结扎冠状动脉左前降支探讨冠心病痰瘀互结证大鼠模型的建立与评价

Establishment and evaluation of phlegm and blood-stasis syndromn on coronary heart disease model by high-fat diet combined with ligation of left anterior descending coronary artery in rats

关键词:  冠心病  痰瘀互结证  血瘀  痰浊  动物模型  大鼠

Key words:  coronary heart disease  phlegm and blood stasis syndrome  blood  stasis  phegm animal model  rat

摘要:

[目的]本实验采用高脂饲料喂养联合结扎冠状动脉左前降支的方法模拟冠心病痰瘀互结证病机的发生发展，以期探讨大鼠冠心病痰瘀互结证模型的建立方法和评价标准。[方法]健康SPF级Wistar雄性大鼠，随机分为空白组、假手术组、痰浊组、血瘀组、痰瘀互结组5组。空白组、假手术组、血瘀组采用普通饲料喂养，痰浊组和痰瘀互结组采用高脂饲料喂养。在8周末，对血瘀组和痰瘀互结组行冠状动脉左前降支结扎术，假手术组行只穿线不结扎术。采集超声心动图，检测大鼠血清中血脂指标、心肌酶指标、血流变指标、血小板聚集指标，并评价冠心病痰瘀互结证大鼠模型。[结果]超声变化：与假手术组比较，冠心病痰瘀互结证组左心室射血分数（EF）、左心室短轴缩短率（FS）、舒张期室间隔厚度（IVSd）、收缩期室间隔厚度（IVSs）、收缩末期左心室后壁厚度（LVPWs）、左心室质量（LV M）显著降低，收缩末期左心室内径（LVIDs）、收缩末期左心室容积（LV Vols）显著升高（P<0.05，P<0.01）；血脂变化：与假手术组比较，痰瘀互结证组总胆固醇（TC）、低密度脂蛋白（LDL）显著升高，高密度脂蛋白（HDL）显著降低（P<0.01）；心肌酶变化：与假手术组比较，痰瘀互结证组肌酸激酶（CK）、肌酸激酶-MB型同工酶（CM-MB）和乳酸脱氢酶（LDH）显著升高（P<0.01）；血流变变化：与假手术组比较，痰瘀互结证组BV1/s、BV5/s、BV30/s、BV200/s和PV显著增高（P<0.05，P<0.01）；血小板聚集变化：与假手术组比较，痰瘀互结证组血小板聚集率（PAR）、最大血小板聚集率（MPAR）显著升高（P<0.01）；心脏组织苏木精-伊红（HE）染色、2，3，5-氯化三苯基四氯唑（TTC）染色：痰瘀互结组大鼠心肌纤维断裂，心肌纤维排列不规则，结构紊乱，肌纤维空隙较宽，间质水肿，大量炎性浸润、坏死，可见明显白色梗死区域。[结论]高脂饲料喂养联合冠状动脉左前降支结扎的方法可以成功建立大鼠冠心病痰瘀互结证病证模型。

Abstract:

[Objective] This experiment used high-fat diet feeding combined with ligation of the left anterior descending coronary artery to simulate the pathogenesis of phlegm and blood-stasis syndromn of coronary heart disease (CHD), in order to explore the establishment of rat model of phlegm and blood-stasis syndromn of CHD.[Methods] Healthy SPF Wistar male rats were randomly divided into control group, sham group, phlegm group, blood-stasis group, and phlegm and blood-stasis group. The control group, sham group, and blood-stasis group were fed with normal diet. Both phlegm group and blood-stasis group were fed with high-fat diet. At the end of the 8th week, the left coronary artery ligation were performed on the blood-stasis group and the phlegm and blood-stasis group. The sham group was performed by opening the chest without with ligation. To collect echocardiogram, detect serum lipids, myocardial enzyme index and hemorrheology index, platelet aggregation index in rats serum; meanwhile, to measure and observe HE staining and TTC staining of heart tissue to evaluate the establishment of rat model of phlegm and blood-stasis syndrome of CHD.[Results] Ultrasound changes:compared with the sham operation group, EF, FS, IVSd, IVSs, LVPWS, LV M were significantly decreased, LVIDs and LV Vols were significantly increased (P<0.05, P<0.01) in phlegm and blood stasis syndrome group. Blood lipid changes:compared with the sham operation group, TC and LDL were significantly increased in the phlegm and blood stasis syndrome group, and HDL was significantly decreased (P<0.01). Myocardial enzyme changes:compared with the sham operation group, CK and CM-MB and LDH were significantly increased in the phlegm and blood stasis syndrome group (P<0.01). Hemorheological changes:compared with the sham operation group, BV1/s, BV5/s, BV30/s, BV200/s and PV were significantly increased in the phlegm and blood stasis syndrome group (P<0.05, P<0.01). Platelet aggregation changes:compared with the sham operation group, the PAR and MPAR of the phlegm and blood stasis syndrome group were significantly increased (P<0.01); HE staining, TTC staining:In the phlegm and blood stasis syndrome group, there were breaked and disorderly myocardial fiber, ande the muscle fiber gaps got wider, full of massive swelling mesenchym, inflammatory and necrotic cells.[Conclusion] It suggested the method that is high-fat diet feeding combined with left anterior descending coronary artery ligation can successfully establish the rat model of Phlegm and blood-stasis syndrome of CHD.