Study On The Effects And Mechanism Of Tongluo Decoction To Myocardial Apoptosis Following Coronary Microembolization In Rats

Objective: To establish coronary artery microembolization model in rats by injecting autologous thrombus through the right common carotid artery by interventional method,and to study the effects and mechanism of Tongluo Decoction on myocardial apoptosis in rats with coronary artery microembolization.Methods: 1.Autologous thrombus model compared with micro embolization ball group: 36 male Wistar rats were randomly divided into blank group(n=6),sham operation group(n=10),autologous thrombosis group(n=10),microembolization ball group(n=10).The 0.5ml suspension autologous thrombus and microembolization spheres were injected into the root of aorta of rats by interventional method,respectively.A autologous thrombus model and a micro embolization ball model of coronary artery microembolization were established in s rats.After 24 h,the cardiac function,the number of microembolization vessels,the area of myocardial ischemia were detected in each group.2.The quality control of Tongluo Decoction with high performance liquid chromatography : Tongluo Decoction concentrated solution was analyzed by high performance liquid chromatography.The control solution and the sample solution of the preparation of calycosin glycoside,paeoniflorin and ginsenoside Rb1 were prepared respectively.The standard curve was made according to the reference substance,and the precision test,repeatability test,stability test,recovery rate test and the content of the sample were determined.3.Tongluo Decoction intervened myocardial apoptosis in rats with coronary microembolization: 46 male Wistar rats were divided into followed groups randomly,blank group(n=6),autologous thrombosis model group(n=10),Tongluo Decoction low-dose group(n=10),Tongluo Decoction hig-dose group(n=10),Tongxinluo group(positive control,n=10).The interventional method was used to establish the coronary microembolism model of rats with autologous thrombus.Tongluo Decoction low-dose group and Tongluo Decoction high-dose group were given low and high dose Tongluo Decoction,Tongxinluo group was treated by Tongxinluo.After 12 days of intervention,the cardiac function,the number of microembolization vessels,the area of myocardial ischemia and the level of cardiomyocyte apoptosis were detected in each group.Results:1.After establishing the CME model,autologous thrombus group and microembolism ball group microembolization vessel count ratio increased,ischemia myocardial area ratio increased(P﹤0.05).Compared with microembolism ball group,autologous thrombus group and microembolism ball group microembolization vessel count ratio increased,myocardial ischemia area ratio increased(P﹥0.05).2.The results of the study showed that the fractions of Radix Astragali,radix paeoniae rubra and red ginseng in Tongluo Decoction were well separated and had no interference with other components.In Tongluo Decoction,calycosin glycoside content is about 75.67 μg/g,paeoniflorin content is about 5432.33μg/g,ginsenoside Rb1 content is about 350.59μg/g.3.Compared with model group,Tongluo Decoction low dose group,Tongluo Decoction high dose group and Tongxinluo group microembolization vessel count ratio、ischemic myocardial area ratio、apoptosis rate decreased(P﹤0.05),bcl-2 protein expression、bcl-2/Bax ratio increased,Bax、caspase-3 protein expression decreased(P ﹤ 0.05);Tongluo Decoction low dose group heart function improved(P﹥0.05),Tongluo Decoction high dose group and Tongxinluo group heart function improved significantly(P﹤0.05).Compared with Tongxinluo group,the effect of Tongluo Decoction low dose group and high dose group on heart function did not change significantly(P﹥0.05),ischemic myocardial area ratio decreased significantly.Tongluo Decoction high dose group microembolization vessel count ratio、cardiomyocyte apoptosis rate and bcl-2、bcl-2/Bax ratio decreased significantly,to compared with Tongluo Decoction low dose group,but the difference was not statistically significant(P > 0.05).Tongluo Decoction low dose group,high dose group bcl-2、bcl-2/Bax ratio was slightly increased,(P﹤0.05);Tongluo Decoction high dose group Bax、caspase-3 protein expression decreased,but the difference was not statistically significant(P > 0.05).Conclusion: 1.The CME model of rats can be successfully established by injection of microspheres through catheter intervention as well as autologous thrombus,but the autologous thrombus model is better than microembolization ball model.2.Tongluo Decoction may inhibit the cardiomyocyte apoptosis of CME rat by blocking mitochondrial apoptosis pathway and improve cardiac function.