Atrial Electrophysiology And Cav1.2/CaM/CaMKâ…¡ Signal Channel Research Of Qi Po Sheng Mai Granule For The Treatment Of Atrial Fibrillation

Objective:To explore Calcium overlord mechanism of atrial fibrillation (Af) electric remodeling in rats with Af and changes on ion channel protein and Calcium-handling-related protein of Cardiac atrium myocyte. To investigate the crucial role during atrial remodeling of Af and find the effective treatment target of QPSM.Method:1. Method of establishing Atrial fibrillation model and atrial electrophysiology:AF models was optimized through 3 different concentration (Mode11: Ach99ug/mL CaCl210mg/mL for 35 days; Model2:Ach66ug/mL CaCl210mg/mL for 35days; Model3:Ach99ug/mL CaCl210mg/mL for 7 days) and each group has 8-10 rats. All rats are traced with electrocardiograph and measured Atrial Effective response period (AERP) by S1S2 programmed electrical stimulation(PES).2. Method of QPSM against Af pharmacodynamics and AERP researchAll rats are randomly seperated into control group(n=10) and model group(n=80) which give tail intravenous injection of Ach99ug/mL CaC1210mg/mL for 7 days.After checking the ECG of the model group to see if it has Af ECG, we seperated all 80 rats in to model group(Model)、Wenxin granule group(WXG)、verapamil group(Verap)、 high does Qiposhengmai granule of high does (High dose)、middle does Qiposhengmai granule of high does (Middle dose)、low does Qiposhengmai granule of high does (low dose),与nomal group (Control).Give medicine for 4 weeks and give tail intravenous injection of Ach99ug/mL CaC1210mg/mL through the 4 weeks. After 4 weeks,anaesthetized rats with pentobarbital sodium, record the ECG and use Langendorff system to get rats atrial effective period.Extract Af model rats atrial protein.Use western blot to detect Cav 1.2、CAM、CaMKⅡ、RYR2、P-RYR2、SK2、 Cx40 protein. Use Rt-PCR to detect Cav1.2、Cx40mRNA.Results:1、Result of establishing Atrial fibrillation model and atrial electrophysiology:1.1 Successful rate of different group after 7 days is:1st Model group is 100%, 2nd Model group is 60%.1.2 Af time of different group after 7 days is:compared 1st Model group and 2nd Model group,1st Model group and normal group,2nd Model group and normal group,p<0.05.1.3 Af time of different group after 35 days is:compared 1st Model group and 2nd Model group,1st Model group and normal group,p<0.05.1.4 AERP of different group is:after 35 days,compared 1st Model group and 2nd Model group, st Model group and normal group, p<0.05.2 pharmacodynamics Results of QPSM granule’effects on Af time and AERP: After 35 days,compared the Af time results of MX and ZC,MX and WLPM,MX and WXQMX and HD,MX and HD,MX and LD, p<0.05<, After 35 days,compared the AERP results of MX and ZV,MX and WLPM,MX and WXQMX and HD,MX and HD,MXandLD, p<0.05.3、Research of QPSM against Atrial Cavl.2/CaM/CaMKⅡ signal channel change in Af.3.1 QPSM granule has effects on Cavl.2 protein and CaV1.2mRNA of Af model rats.3.2 QPSM granule has effects on CaM protein of Af model rats.3.3 QPSM granule has effects on CaMKII protein of Af model rats.3.4 QPSM granule has effects on RyR2 protein and phosphorylation RyR2 protein of Af model rats.3.5 QPSM granule has effects on SK2 protein of Af model rats.4、Research of QPSM against Atrial Cx40 protein and mRNA change in Af.4.1 QPSM granule has effects on Cx40 protein and Cx40mRNA of Af model rats.4.2 QPSM granule has effects on ultrastructure of Af model rats.Conclusion:1.. It has high successful rate to establishing Atrial fibrillation model by tail intravenous injection of Ach99ug/mL CaC1210mg/mL for 35 days. Atrial fibrillation model had a significant change of electric and calcium handling related protein.2n QPSM granule regulate calcium handling during AF by giving effects on Af time and AERP of Atrial fibrillation model by shortening Af time and prolong AERP.3、QPSM granule has effects onCav 1.2, CaM, CaMKⅡ, Cav1.2, RyR2 and RYR2 phosphorylation.