Mechenism Of Shenfu Injection Regulation On Ion Channels Of Heart Failure Myocyte

Objective Shenfu injection,a traditional Chinese medicine compound preparations from shenfu soup,which is made up of Radix Ginseng and Radix Aconiti Lateralis,Shenfu injection can treatment shock,arrhythmia and heart failure in clinic.This experiment made the chronic heart failure(CHF)models of SD rats,and studied shenfu injection how to enhancing cardiac contractility and improving heart function from the whole animal,cells and the ultrastructure level.In order to providing new theoretical basis of shenfu injection used in heart failure treatmentMethods(1)This experiment was divided into three group:sham operated rats,chronic heart failure rats and shenfu injection treats group(rats with 6ml/kg shenfu injection),each group had 20 rats.in cell experiment,shenfu injection set 1.25%,2.5%,5%and 10%concentration gradient.(2)CHF models were induced by permanent ligation on the left anterior descending coronary artery.Chronic heart failure rats and shenfu injection treats group were used CHF model rats,sham operated rats were the same procedure without ligation of the LAD.Regular dieting four weeks later,heart failure indicators were assessed.Ultrastructures of myocardium were observed by transmission electron microscope.(3)The measurement of the ICa-L peak amplitude and SR Ca2+load:Single rat ventricular cardiomyocytes were enzymatically isolated through Langendorff apparatus.A whole-cell patch clamp technique was employed to record ICa-L in ventricular myocytes.With fluorescent indicator Fluo-3/AM dying myocytes,we monitored SR Ca2+load as the change in Fluo-3/AM fluorescence level after depletion of the SR with caffeine using confocal microscopy.(4)SPSS21.0 software was used for the statistical analyses.Datum were expressed as mean±SD for all experimental results.Statistical analysis were made with one way-ANOVA and followed by LSD-t for individual comparisons of means.P-values of<0.05 were considered significant.Results(1)Echocardiography showed that LVIDd and LVIDs was obviously increased(P<0.01),LVPWd/LVPWs,LVAWd/LVAWs,EF and FS were remarked decreased(P<0.01)in CHF group compared with the sham operated group;LVIDd and LVIDs was obviously decreased(P<0.01),LVPWd/LVPWs,LVAWd/LVAWs,EF and FS were remarked increased(P<0.01)in shenfu injection treats group compared with the CHF group.(2)Effects of shenfu injection on ultrastructure of myocardium:myofilaments in myocardium of the sham operated group were lined up in line,structures of mitochondria were intact and cristae were clear.The ultrastructure of myocardium in the CHF rats displayed the myofilaments were dissolved,and a lot of myofilaments were disordered,in addition,some cristae were dissolved and their outlines were vague.In shenfu injection treats rats,myofilaments ranked regularly and had mild dissolved,structures of mitochondria were complete and most of cristae were clear.(3)Effects of shenfu injection on L-type calcium currents of myocardial cells:compared with the sham group,The intensity of L-type calcium currents of Myocardial cells was remarked decreased in CHF group(p<0.05).Shenfu injection could reinforced the L-type calcium currents in a concentration-dependent manner(1.25%,2.5%,5%,10%)compared with the CHF group.The L-type calcium currents between each concentration had significant strength increases(p<0.05)except the concentration of 5%and 10%.Moreover,compared with the sham group,Shenfu injection also could reinforced the L-type calcium currents in a concentration-dependent manner(1.25%,2.5%,5%,10%),and each concentration had significant strength increases(p<0.05)except the concentration of 5%and 10%.(4)Effects of shenfu injection on SR Ca2+ load:the fluorescence of cardionyocytes stained with Fluo-3/AM were dramatically reduced in the CHF group,and spatial averages of Ca2+transients(? F/F0)of the CHF group were reduced markedly compared with the sham group(p<0.05).(5)Shenfu injection could reinforced the fluorescence of cardiomyocytes in a concentration-dependent manner(1.25%,2.5%,5%,10%)compared with the CHF group.The fluorescence of cardiomyocytes between each concentration had significant strength increases(p<0.05)except the concentration of 5%and 10%.Moreover,compared with the sham group,Shenfu injection also could reinforced the fluorescence of cardiomyocytes in a concentration-dependent manner(l.25%,2.5%,5%,10%),and each concentration had significant strength increases(p<0.05)except the concentration of 5%and 10%.Conclusion In conclusion,Shenfu Injection attenuates cardiac dysfunction of rat failing hearts,our data identify a novel mechanism that enhanced amplitude of ICa-L,preserved SR Ca2+content and restored ultrastructure contribute to improving heart function,adding new evidence of Shenfu Injection to improve therapeutic options of HF patients based on mechanistic insights.