The Protective Effect And Potential Mechanisms Of Danlou Pian On Coronary Heart Disease With Phlegm And Blood Stasis In Rats

Objective: To research the protective effect of Danlou Pian on coronary heart disease withphlegm and blood stasis in rats and explore its potential mechanisms.Methods:210Wistar rats, male and female half, were randomly divided into seven groupsand there were30rats in each group. The Control group was fed normally. Other groups werefed high fat diet and administered orally with fat emulsion. From the twenty-first day byfeeding high fat diet, the Control group was administered orally with distilled water.Shamgroup and Model group were administered orally with0.5%Sodium carboxymethyl cellulose.Danlou pian groups were administered orally with Danlou pian500mg/kg/d,1000mg/kg/d,2000mg/kg/d each. Isosorbide Dinitrate group were administered orally with IsosorbideDinitrate tablet3mg/kg/d. once a day. From the thirty-five day, ligature the coronary artery forthe High fat diet groups. The Sham group wore the line only.After the operation, intramuscularinjected penicillin for three days. Feeding high fat diet till the forty ninth day, take the bloodand hearts to investigate: ESR, EET, FC, PAR, PAG, MPAG, BV, PV, HCT, TC, TG, HDL-c,LDL-c, CK-MB, AST, LDH and MIS; western blot to investigate the Procaspase-3,Procaspase-9, Bax, Bcl-2protein expression; QPCR to investigate the Bax, Bcl-2mRNAexpression; investigate the Caspase-3, Caspase-9activity; observe the morphological changesin myocardial tissue and myocardial ultrastructural changes.Results:1. Compared with the Control group, ESR, EET, PAR, FC, HCT, EAI, IR, PAG at1min,PAG at3min, PAG at5min, MPAG,20/s BV,60/s BV,120/s BV and120/s PV in the Shamgroup were significantly increased (P<0.05,P<0.01or P<0.001);Compared with the Shamgroup, ESR, EET, PAR, FC, HCT, EAI, IR, PAG at1min, PAG at3min, PAG at5min, MPAG,20/s BV,60/s BV,120/s BV and120/s PV in the Model group were significantly increased(P<0.05orP<0.01); Compared with the Model group, ESR, EET, PAR, FC, HCT, EAI, IR, PAGat1min, PAG at3min, PAG at5min, MPAG,20/s BV,60/s BV,120/s BV and120/s PV in the DLP and IDT groups were significantly reduced (P<0.05orP<0.01).2. Compared with the Control group, TG content, TC content, LDL-c content, AI and CRIin the Sham group and Model group were significantly increased, HDL-c content weresignificantly reduced (P<0.05or P<0.01); Compared with the Model group, contents of TG, TC,LDL-c, and AI, CRI in the IDT group had no effect, but the DLP1000,2000mg/kg groupswere significantly reduce, content of HDL-c were significantly lower (P<0.05or P<0.01).3. Compared with the Sham group, activities of LDH, AST and CK-MB in the Modelgroup were significantly increased (P<0.01), Compared with the Model group, activities ofLDH, AST and CK-MB in the IDT and DLP1000,2000mg/kg groups were significantly lower(P<0.01).4. Compared with the Control group, activities of Caspase-3, Caspase-9in the Sham groupwere significantly increased (P<0.01), Compared with the Sham group, activities of Caspase-3,Caspase-9in the Model group were significantly increased (P<0.01); Compared with the Modelgroup, activities of Caspase-3, Caspase-9in the IDT and DLP1000,2000mg/kg groups weresignificantly lower (P<0.05or P<0.01).5. Compared with the Control group, Procaspase-3protein expression, Procaspase-9protein expression and Bcl-2protein expression in the Sham group were significantly reduced(P<0.05or P<0.01), Bax protein expression in the Sham group were significantly increased(P<0.001); Compared with the Sham group, Procaspase-3protein expression, Procaspase-9protein expression and Bcl-2protein expression in the Model group were significantly reduced(P<0.05or P<0.01), Bax protein expression in the Model group were significantly increased(P<0.05); Compared with theModel group, Procaspase-3protein expression, Procaspase-9protein expression and Bcl-2protein expression in the IDT and DLP1000,2000mg/kg groupswere significantly increased (P<0.01orP<0.001),Bax protein expression in the IDT and DLP1000,2000mg/kg group were significantly reduced (P<0.05or P<0.01).6. Compared with the Sham group, relative Bax mRNA levels in the Model group weresignificantly increased, relative Bcl-2mRNA levels in the Model group were significantlyreduced (P<0.01); Compared with the Model group, relative Bax mRNA levels in the IDT and DLP1000,2000mg/kg groups were significantly reduced, relative Bcl-2mRNA levels in theIDT and DLP1000,2000mg/kg groups were significantly increased (P＜0.05or P<0.001).7. Compared with the Control group, MIS in the Sham group were significantly increased(P<0.001), Compared with the Sham group, MIS in the Model group were significantlyincreased (P<0.001); Compared with theModel groupMIS in the IDT and DLP500,1000,2000mg/kg groups were significantly lower (P<0.05or P<0.01).8. Compared with the Control group, TUNEL grayscale in the Sham group weresignificantly lower (P<0.001), Compared with the Sham group, TUNEL grayscale in the Modelgroup were significantly lower (P<0.001); Compared with the Model group TUNEL grayscalein the IDT and DLP1000,2000mg/kg groups were significantly increased (P<0.05or P<0.01).9. Compared with the Sham group, myocardial tissue and cardiac myocyte of the Modelgroupwere significantly pathological damaged; Compared with the Model group, IDT and DLP500,1000,2000mg/kg groups reduced pathological damages of myocardium and myocardialcells.Conclusion:1. Danlou Pian can significantly reduce the MIS, the activities of LDH, AST and CK-MB,reduced pathological damage of myocardium and myocardial cells in coronary heart diseasewith phlegm and blood stasis rats. It has obvious protective effects on myocardial ischemia.2. Danlou Pian can significantly reduce the blood viscosity to prevent thrombosis incoronary heart disease with phlegm and blood stasis rats.3. Danlou Pian can improve the disorder of lipid metabolism in coronary heart diseasewith phlegm and blood stasis rats.4. Danlou Pian can significantly reduce Caspase-3activity, Caspase-9activity Bax proteinexpression, Bax mRNA levels and increase Bcl-2protein expression，Procaspase-3proteinexpression, Procaspase-9protein expression, Bcl-2mRNA levels and the TUNEL grey level incoronary heart disease with phlegm and blood stasis rats. It proves that Danlou Pian can inhibit apoptosis of myocardial cells.Above all, Danlou Pian has a protective effect on myocardial infarction and phlegm andblood stasis in rats. The possible mechanisms are: reduce blood viscosityto prevent thrombosisand inhibit apoptosis of myocardial cell.