An Animal Experimental Study On The Effect Of Naoxintong Jiaonang Combined With Dual Antithrombotic Therapy After Coronary Microembolization In Rats

ObjectiveThe aim of this study was to establish a model of coronary microembolization(CME) in rats to observe the quantity of thrombus in coronary microembolization, myocardial apoptosis rate and the balance between bleeding and clotting after combined therapy with Naoxintong Jiaonang and dual antithrombotic therapy (clopidogrel+aspirin), compared with dual antithrombotic therapy and Naoxintong Jiaonang therapy on CME, and to discuss the advantagement of combined therapy with Naoxintong Jiaonang and dual antithrombotic therapy, to provide an experimental evidence for adjusting the disorder caused by dual antithrombotic therapy.MethodsWe created a rat model of CME by injecting a suspension of microthrombotic particles into left ventricle. The microthrombotic particles were generated from the rat clots. Ninety-five rats were randomly divided into six groups on the basis of the result of preliminary experiment:control group (CL group, n=10), Sham-operation group (SO group, n=10),CME model group(CME group, n=15),and Naoxintong Jiaonang intervention group (NJ group, n=15),Dual Antithrombotic therapy intervention group(DA group, n=30), combined therapy with Naoxintong Jiaonang and Dual Antithrombotic intervention group(NDA group, n=15). The remaining sixty-nine rats obtained complete data (CL group=10,SO group =10,CME group=11,NJ group=11,DA group=13 and NDA group=14).The animals were preadiministered for 3 days and continued for 1 week following the procedures, and then were sacrificed for the blood and heart preparations. HE staining was used to assess microthrombus in the coronary microcirculation. TUNEL staining and immunohischemical method for caspase-3 were employed to calculate myocardial apoptosis rate. Tail-cutting method was conducted to assess the bleeding time and glass-slide method to detect the clotting time, and turbidimetry to minitor alteration of platelet aggregation rate.Result1. Validation of model of CMECompared with SO group, both the bleeding time and the clotting time were decreased in CME group(P<0.05 or P<0.01), while the quantity of thrombus in coronary microembolization, myocyte apoptosis rate and platelet aggregation rate were increased(P<0.01). 2. Effects of drug intervention on the quantity of thrombus and myocardial apoptosis rate in coronary microembolizationCompared with CME group, the quantity of thrombus in coronary microembolization was decreased in NJ group,DA group and NDA group(29.04%±1.85% vs.25.65%±1.92%,21.03%±1.82%,18.80%±1.97%, P<0.01); Compared with DA group and NJ group, the indicators were decreased in NDA group(P<0.01).Compared with CME group, myocardial apoptosis rates were decreased in NJ group,DA group and NDA group(P<0.01); Compared with DA group and NJ group, the indicators were decreased significantly in NDA group(P<0.01).3. Effects of drug intervention on the balance between bleeding and clottingCompared with DA group, NDA group could inhibit the incidence of surgical bleeding significantly(P<0.0083).Compared with CME group, both the bleeding time and the clotting time were increased in NJ group,DA group and NDA group(P<0.01); Compared with NJ group and NDA group, these indicators were increased in DA group (P<0.05 or P<0.01).Compared with CME group, different concentrations of ADP-induced maximum platelet aggregation rate were decreased in NJ group,DA group and NDA group(P<0.01); Compared with NJ group and NDA group, these indicators were decreased significantly in DA group(P<0.01).ConclutionsCombined therapy with Naoxintong Jiaonang and dual antithrombotic therapy could not only decrease the quantity of thrombus and myocardial apoptosis rate in coronary microembolization, but also maintain the balance between bleeding and clotting, decrease the side effect of bleeding by antithrombotic therapy, which maybe provide an experimental evidence for adjusting the disorder caused by dual antithrombotic therapy.