| Coronary heart disease is a severe disease to the adult,which has very high incidence and death rate in many countries and regions.Therefore,the development of drugs for coronary heart disease has been considered as the priority all over the world.More and more attention has been paid to the traditional Chinese medicine with regard to protecting coronary heart disease.To study coronary heart disease,it is necessary to establish an ideal animal model. Chinese experimental miniature swine were used to establish the disease-syndrome integrated models with the syndrome of phlegm and stagnated blood by high fat diet with coronary artery injury in the Experiment Research Center of Xiyuan Hospital,the pathological changes of the models are similar to Man's.To study the rule and nature of Fang-Zheng correlation theory,the model animals had been fed with three sorts of traditional Chinese medicinal formulae,which were Gualou Xiebai Banxia Tang as Hua Tan Fang,Xuefu Zhuyu Tang as Qu Yu Fang and Gualou Xiebai Banxia Tang plus Xuefu Zhuyu Tang as Tan Yu Tong Zhi Fang.Based on the diseased animal models and experimental researches,the current research aims to observe the histomorphology and structure of coronary artery injured and infracted myocardium,and to study the effects of traditional Chinese medicinal formulae.The present paper also investigated the change of myocardial apoptosis cells and expression of Bax,bcl-2,Caspase-3 and Caspase-9 of each group during the disease and the effect of traditional Chinese medicinal formulae was intensively evaluated.Materials and MethodsThirty Chinese experimental miniature swine(CEMS) were randomly divided into six groups:(1)Control group(Con),ordinary diet without coronary artery injury;n=5;(2)Phlegm and stagnated blood model group(Model),high fat diet with coronary artery injury,n=5; (3)Western Medicine group(XY),high fat diet with coronary artery injury plus Simvastatin 0.5mg/kg,n=5;(4)Hua Tan group(HT),high fat diet with coronary artery injury plus Guolou Xiebai Banxia Tang,6g raw herb/kg,n=5;(5)Qu Yu Group(QY),high fat diet with coronary artery injury plus Xuefu Zhuyu Tang,8g raw herb/kg,n=5;(6)Tan Yu Tong Zhi group(TY), high fat diet with coronary artery injury plus Guolou Xiebai Banxia Tang,3g raw herb/kg plus Xuefu Zhuyu Tang,4g raw herb/kg,n=5.After the surgical operation and on the second day, the animals were continuously fed with high fat diet for 8 weeks.The coronary artery injured and infracted juncture myocardium was made into paraffin section after the experiment was over.The histomorphology and structure on coronary artery injured was observed through the HE stain,EVG stain and computer imagine analysis,and the detected data were analyzed with the software of the SPSS 11.5.The histomorphology and structure on infract juncture myocardium was analyzed through the HE stain and Masson stain.To investigate the Myocardial apoptosis cells of each group,situ TDT-mediate dUTP nick end labeling(TUNEL),bax,bcl-2,caspase-3 and caspase-9 in immunohistochemical staining were introduced into the study.Meanwhile we study the effect of traditional Chinese medicinal formulae on myocardial apoptosis cells and the expression ofbax,bcl-2,caspase-3 and caspase-9 in the diseased animal models.Results1.Compared with the control group,the lumens of coronary artery had eccentrically narrow to some different extent in the model group with significant difference(P＜0.01).The lumens of coronary artery were narrow in each treatment group.But compared with the model group,the very significant difference existed in HT group,QY group and TY group(P＜0.01), the significant difference existed in XY group(P＜0.05).2.Cardiac muscle fibers in model group were showed to be broken and lost in a lamellar manner,and the region was replaced with fibrous tissue,with focus cardiac muscle fiber degeneration and necrosis,fatty infiltration and inflammatory cell infiltration.Compared with the control group,the very significant difference existed in model group(P＜0.01).Compared with the model group,the cardiac muscle pathological changes had significant relief,the very significant difference existed in HT group and XY group(P＜0.01),the significant difference existed in QY group and TY group(P＜0.05).3.When the coronary artery injured was analyzed through the computer imagine analysis, the results were as follow:the lumens of coronary artery became significantly narrow (P＜0.01),and the max intimate thickness of the coronary artery have been obviously increased in the model(P＜0.01),compared with the control group.The proportion of the max intimate and tunica media thickness in model group was significantly larger than that of control group(P＜0.01).The narrow degree of coronary artery in each treatment group was significantly lighter than that of model group(P＜0.01).The max intimate thickness of the coronary artery in each treatment group was significantly less than that of model group (P＜0.01).The proportion of the max intimate and tunica media thickness in each treatment group was significantly less than that of model group(P＜0.01).4.The results of immunohistochemistry showed that light density ofbcl-2 and bcl-2/bax in model group was significantly less than that of control group(P＜0.01),light density of bax in model group was significantly more than that of control group(P＜0.01).The results showed that light density of bcl-2 and bcl-2/bax in TY group was significantly more than that of model group(P＜0.01),light density ofbax in TY group was significantly less than that of model group(P＜0.01).5.The positive expressions of caspase-3 and caspase-9 in model group were significantly more than that of control group(P＜0.01),The positive expressions of caspase-3 and caspase-9 in TY group were significantly less than that of model group(P＜0.01).6.Myocardial apoptosis index of each group was determined by TUNEL,the results showed that apoptosis indexes in control group,model group and TY group were respectively 0.92%,27.68%and 17.28%.The numbers of positive Myocardial cells and myocardial apoptosis index in model group and TY group were significantly more than these of control group(P＜0.01).The results showed that the numbers of positive myocardial cells and Myocardial apoptosis index in model group were significantly more than these of TY group (P＜0.01).Conclusion1.It was confirmed through pathological methods that the establishment of Chinese miniature swine model with coronary heart diseases and the syndrome of phlegm and stagnated blood was successful.2.The coronary heart diseases and the syndrome of phlegm and stagnated blood in the Chinese miniature swine model could be prevented and cured effectively by the traditional Chinese medicinal formulae.3.The ischemic myocardial apoptosis cells in the Chinese miniature swine model could be protected effectively by the traditional Chinese medicinal formulae.
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