Effects Of Kangxian Yixin Recipe On NE-induced Cardiomyocyte Hypertrophy And Apoptosis Factors Bcl-2 And Bax

Dilatedcardiomyopathy(DCM)is a heterogeneous heart disease disease characterized by unilateral or bilateral ventricular enlargement,often accompanied by progressive myocardial systolic dysfunction.DCM is the main cause of end-stage heart failure,and ventricular remodeling is the main pathological process of DCM.There are hypertrophy,degeneration and necrosis of cardiac myocytes in the cardiac tissue of DCM rats,and these pathological changes are related to the excessive increase of sympathetic nerves.Research shows that the Autonomic nervous system(ANS)neurotransmitter norepinephrine(NE)binds to adrenergic receptors,activating the sympathetic nervous system and inducing hypertrophy and apoptosis of cardiac muscle cells.Kangxian Yixin Decoction(KYD)is a clinical empirical prescription for the treatment of DCM,which has an obvious therapeutic effect on patients with DCM type of qi deficiency and blood stasis.Previous studies have proved that KYD can regulate the cardiac function of DCM rats,inhibit ventricular remodeling and reduce apoptosis in myocardial tissue,but the mechanism remains unclear.Therefore,the study on the effect of KYD on NE induced hypertrophy and apoptosis of myocardial cells has important theoretical and clinical significance for the prevention and treatment of central ventricular remodeling and heart failure during the development of DCM,as well as the systematic and objective evaluation of anti-fibrinolytic heart prescription.Objective: H9c2 rat myocardial cell model was used to investigate the effect of KYD on NE induced hypertrophy and apoptosis of myocardial cells.Methods:1.H9c2 rat myocardial cell culture.2.Preparation of KYD and NE.3.QPCR was used to screen the optimal intervention concentration and time for NE induced myocardial hypertrophy and apoptosis.4.Cck-8 was used to detect the effect of KYD on myocardial cell activity.5.Observe the effect of KYD on myocardial hypertrophy induced by NE under the microscope.6.QPCR was used to detect the expressions of ANP and BNP mRNA,apoptosis-related factors bcl-2,Bax and Caspase 3 mRNA in myocardial hypertrophy.7.TUNEL staining was used to observe the occurrence of apoptosis of myocardial cells.8.Flow cytometry was used to detect the apoptosis rate of cardiac myocytes.9.Western Blot was used to detect the expressions of apoptosis-related proteins bcl-2,Bax and Caspase 3 in cardiac myocytes.Results:1.100 ?M NE acted on H9c2 myocardial cells for 24 h,which was the best condition to induce myocardial hypertrophy.2.The 0.25 mg /ml dose of KYD has no cytotoxicity,and can effectively antagonize the occurrence of cardiac hypertrophy and apoptosis.3.QPCR results showed that anti-fiber and beneficial heart could effectively reduce the ANP and BNP mRNA of cardiac myocyte hypertrophy indicators,down-regulate the Bax and Caspase 3 mRNA of apoptosis indicators,and up-regulate the bcl-2 mRNA.4.The results of TUNEL staining showed that KYD could inhibit the apoptosis of myocardial cells induced by NE.5.The apoptosis rate of myocardial cells detected by flow cytometry showed that KYD could significantly reduce the apoptosis rate of neinduced myocardial cells.6.Western Blot results showed that anti-fibrinolysis could significantly regulate ne-induced apoptosis-related proteins bcl-2,Bax and Caspase 3.Conclusion:1.The occurrence of hypertrophy of cardiac myocytes could be induced by the action of 100 ?M NE on H9c2 cardiac myocytes for 24 h.2.The 0.25 mg /ml KYD significantly antagonised ne-induced hypertrophy and apoptosis of H9c2 myocardial cells.3.KYD regulates the expressions of ANP,BNP,Bcl2,Bax and Caspase 3,so as to regulate the hypertrophy of cardiac myocytes and the apoptosis of hypertrophic cardiac myocytes.