| Heart failure is the end stage of various cardiovascular diseases,which threatens human health seriously.Multiple studies have shown that heart failure is as malignant as tumors.Less than half of patients whose heart function were grade ? or ?(New York heart function grading)can survive more than 5 years.The most important cause of death is sudden cardiac death caused by various malignant arrhythmias.Therefore,reducing the incidence of arrhythmia is an important measure to reduce the degree of malignant heart failure and improve the survival rate of cardiac patients.Modern medical treatment of heart failure arrhythmia is mainly based on chemical drugs,but many studies have shown that almost all antiarrhythmic dugs can cause arrhythmia,and even increase the mortality rate ultimately.Inhibition of sodium current remodeling is a hot spot in the research of antiarrhythmic drugs in recent years.In heart failure,phosphorylation of sodium channels due to the activation of protein kinases such as CaMK? can cause an increase in late sodium current which is one of the pathological mechanisms of ventricular arrhythmia in heart failure.The electrocardiogram is characterized by prolongation of QT interval.In the traditional Chinese medicine theory,qi and blood,yin and yang deficiency is the main reason causing heart failure.These pathological factors are often accompanied by phlegm and blood stasis,which causing restlessness,sharp beats in the heart,panic,expressed as palpitations.The treatment of ventricular arrhythmia based on traditional Chinese medicine has certain advantages.Shenlian Fumai granule(SLFM granule)has the effect of replenishing qi and activating blood circulation,clearting heart and dissipating phlegm,and calming the nerves.It is safe and effective in clinical treatment of ventricular arrhythmia.Our previous clinical trials found that SLFM granule can shorten the action potential prolongation caused by isoproterenol,and the prescription of Codonopsis pilosula can inhibit the expression of CaMK? in heart failure and reduce the incidence of ventricular arrhythmia eventually.Therefore we hypothesize that SLFM granule may exert an antiarrhythmic effect by inhibiting the CaMK? and late sodium current pathway.PurposeTo explore the effect and possible mechanism of SLFM Granule on heart failure arrhythmia.MethodsStudy 1:We made models by contracting thoracic aorta of mice.Mice were divided into sham group,model group,high dose group,middle dose group,low dose group,amiodarone group.After4 weeks intervention,ultrasound,electrocardiogram were detected and arrhythmia was induced by isoproterenol.The intervention and optimal dosage of SLFM granule on heart failure arrhythmia were observed.Study 2:We got cardiomyocytes by isolating hearts of mice in sham,model,middle dose and amiodarone groups acutely.The concentration of Ca2+,Na+ and ROS were detected by different fluorescent probes.The expression of calmodulin was detected by Western Blot.Study 3:We divided mice into sham,model,SLFM and mexiletine groups.After 4 weeks of intervention,ultrasound and ECG parameters were detected.Patch clamps were used to recording myocardial action potential and sodium ion channel.The expression of Nav1.5 and SCN5A were detected by Western Blot and PCR.ResultsStudy11.General condition:There were 12 mice in each group at the beginning.There was no death in sham group during4 weeks.There were 6 deaths in model group,4 deaths in high dose group,low dose group and western medicine group.There were 3 deaths in middle dose group.2.Echocardiography results:the results showed that compared with the sham group,ejection fraction and short-axis systolic rate in the model group were significantly decreased(P<0.01).Compared with the model group,ejection fraction and short-axis systolic rate increased in the medium dose group(P<0.05).3.Electrocardiogram results:Compared with the sham group,the electrocardiogram indicators in the model group were significantly different(P<0.01).Compared with the model group,the RR interval of the high-dose group was prolonged(P<0.01),heart rate decreased(P<0.05),QRS time was prolonged(P<0.01),QT and corrected QT interval were shortened(P<0.01).The middle dose group had longer RR interval(P<0.01),lower heart rate(P<0.01),longer PR interval(P<0.01),longer QRS interval(P<0.01),shorter QT and corrected QT interval.In the low-dose group,RR interval was prolonged(P<0.05)and PR interval was prolonged(P<0.01).In western medicine group,RR interval was prolonged(P<0.01),heart rate decreased(P<0.05),PR interval was prolonged(P<0.01),QRS interval was prolonged(P<0.01),QT interval and corrected QT interval were prolonged(P<0.01).The RR interval was the longest(P<0.01),the heart rate was the slowest(P<0.05),and the QT and QT interval was the shortest(P<0.01).The PR interval was the shortest in the high-dose group(P<0.01),and the QRS duration was the longest(P<0.01).Compared with western medicine group,RR interval,QT interval and corrected QT interval in all traditional Chinese medicine groups were significantly shortened(P<0.01).4.A:rrhythmias:Compared with the sham group,the incidence of premature beats in the model group was significantly increased(P<0.01).Compared with the model group,the incidence of premature beats decreased in each administration group(P<0.01).Compared with the traditional Chinese medicine group,the middle-dose group had the lowest incidence of premature beats(P<0.01).After the inj ection of isoproterenol,the incidence of premature beats was increased in all groups.Compared with the model group,the incidence of premature beats decreased in the medium dose group(P<0.05)and the western medicine group(P<0.01).Compared with western medicine group,premature beats were increased in each group(P<0.01).Study21.Fluorescence detection results:Compared with the sham group,the concentration of calcium ions increased,the concentration of sodium ions decreased,and the ROS content increased in the model group(P<0.01).Compared with the model group,the calcium ion concentration and ROS concentration in the traditional Chinese medicine group and the western medicine group decreased,while the sodium ion concentration increased(P<0.01).2.Western Blot results:Compared with the sham group,SERCA/PLB ratio and NCX expression were significantly decreased in the model group(P<0.01),and CaMKII expression was increased(P<0.01).Compared with the model group,SERCA/PLB ratio and NCX expression level increased in traditional Chinese medicine group and the western medicine group(P<0.05).The expression of CaMKII decreased in the traditional Chinese medicine group(P<0.05).Compared with the western medicine group,the expression of CaMKII in thetraditional Chinese medicine group decreased(P<0.05).Study31.Echocardiography results:Compared with the sham group,ejection fraction(EF)and left ventricular short-axis systolic rate(FS)in the model group were significantly decreased(P<0.01).Compared with the model group,ejection fraction and short-axis systolic rate were improved in the traditional Chinese medicine group and the western medicine group(P<0.01)·Compared with western medicine group,ejection fraction and short-axis systolic rate were improved in trraditional Chinese medicine group(P<0.01).In terms of cardiac structure,compared with the sham operation group,the left ventricular systolic diameter and volunie of the model group were significantly increased(P<0.01),while compared with the model group,the left ventricular systolic diameter and volume of the Chinese medicine group were restored(P<0.05).There was no significant difference in diastolic function between the groups.2.Results of ECG parameters:Compared with the sham group,RR interval was significantly shortened in the model group(P<0.01),heart rate increased(P<0.01),QRS interval was shortened(P<0.01),QT and QT interval were shortened(P<0.01).Compared with the model group,QRS duration was prolonged(P<0.01),QT interval was shortened(P<0.01),QT and corrected QT interval was shortened(P<0.01)in the traditional Chinese medicine group.Compared with the western medicine group,the QRS interval was prolonged in the traditional Chinese medicine group(P<0.01)and the QT interval was prolonged in the western medicine group(P<0.05).3.Action potential results:When the stimulation frequency was 1.0Hz,the action potential duration of the model group was prolonged compared with that of the sham group(P<0.01).Compared with the model group,the action potential duration and recovery time were shortened in the traditional Chinese medicine group(P<0.01),and shortened in the western medicine group(P<0.01),and shortened in the recovery time to 90%(P<0.05).Compared with the western medicine group,the duration of action potential and its recovery to25%,50%and 90%were shortened in the Chinese medicine group(P<0.01or P<0.05).When the frequency was 2.0 hz,the action potential duration of the model group was prolonged compared with that of the sham group(P<0.01).Compared with the model group,the action potential duration and recovery time were shortened in the traditional Chinese medicine group(P<0.01),and shortened in the western medicine group(P<0.01),and shortened in the recovery time to90%(P<0.05)..4.Sodium current results:Compared with the sham group,the model group showed decreased peak sodium current,increased late sodium current,prolonged recovery time,and increased late sodium current ratio(P<0.01).Compared with the model group,the peak sodium current restored,the recovery time shortened,and the late sodium current and ratio decreased in the traditional Chinese medicine group and the western medicine group(P<0.01).After perfusion with KN93,compared with the sham operation group,the late sodium current increased in the model group(P<0.05).Compared with the normal condition,the peak sodium current and late sodium current in the sham operation group decreased significantly,and the recovery time was shortened(P<0.01).In the model group,peak sodium current decreased(P<0.05),late sodium current and ratio decreased,and recovery time decreased(P<0.01).There were significant differences in each index between the traditional Chinese medicine group and the western medicine group(P<0.01).Compared with the western medicine group,there was no significant difference in each index between the Chinese medicine group and the western medicine group.5.Sodium channel inactivation curve results:Compared with the sham operation group,the inactivation curve of the model group was significantly shifted to the left.Compared with the model group,the inactivation curve of the Chinese medicine group and the western medicine group recovered,which was basically consistent with the sham operation group.Compared with the sham operation group,the voltage of the model group was significantly increased(P<0.01)and the slope decreased(P<0.01).Compared with the model group,the intoxication of the Chinese medicine group and the western medicine group was 50%(P<0.01).After blocking KMKII with KN93,the sodium channel inactivation curves of each group moved downward.Compared with the normal case,when KN93 was perfused,the model group inactivated 50%voltage significantly increased(P<0.01),and the slope increased significantly(P<0.01);Chinese medicine group(P<0.01)and western medicine group(P<0.05).The slope increases.6.Recovery curve after inactivation:Compared with the sham operation group,after the model group was inactivated,the recovery curve moved to the lower rnght and the starting point moved upward;compared with the model group,the curve of the traditional Chinese medicine group and the western medicine group recovered,and the starting point moved downward.Compared with the sham operation group,the time constant of the model group increased(P<0.01),and the initial recovery ratio increased(P<0.01).Compared with the model group,the time constant and the initial recovery ratio of the Chinese medicine group and the western medicine group decreased.P<0.01).After perfusion of KN93,the sham operation group and the Chinese medicine group failed to stimulate the recovery current under the stimulation procedure,and the recovery curve after inactivation could not be drawn.After the inactivation of the model group and the western medicine group,the recovery curve was obviously moved down and shifted to the right,and the time constant was significantly increased(P<0.01).7.Sodium channel and CaMKII gene and protein expression results:Compared with the sham operation group,the CaMKII gene(P<0.01),protein and phosphorylated water in the model group were significantly increased(P<0.05),Nav1.5protein and gene expression.Significantly increased(P<0.05);compared with the model group,CaMKII gene(P<0.05),protein(P<0.05),and phosphorylation level(P<0.01)in the Chinese medicine group were significantly decreased,Nav1.5gene and protein expression were significantly decreased.Significantly decreased(P<0.01);the expression levels of Nav1.5gene(P<0.05)and protein(P<0.01)decreased in western medicine group.Compared with the western medicine group,CaMKII gene and SCN5A gene expression were decreased in the Chinese medicine group(P<0.01),and CaMKII phosphorylation level was decreased(P<0.05).Conclusion1.SLFM granules can improve the electrophysiological remodeling of arrhythmia mice with heart failure caused by stress load,inhibit arrhythmia of heart failure,and have no side effect of arrhythmia.2.The inhibitory effect of SLFM granules on arrhythmia may be related to inhibition of CaMK? expression,recovery of sodium channel function,inhibition of late sodium current,and regulation of intracellular ion concentration.3.The usual clinical dosage is the optimal dose. |
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