Epigallocatechin Gallate Improves Cardiomyocytes Hypertrophy By Inhibiting HDAC2

Objective:To investigate the effects of green tea extract epigallocatechin gallate(EGCG)on attenuating the cardiomyocytes hypertrophy induced by phenylephrine(PE)through inhibiting histone deacetylase 2(HDAC2)in mice,and to provide new ideas for clinical prevention and treatment of hypertrophic cardiomyopathy.Methods:1.Myocardial cells of mice were isolated by enzymatic digestion and randomly divided into normal group,phenylephrine(PE)group,and solvent group(dimethylsulfoxide + PE).The three groups were tested as follows:(1)CCK-8 method was used to detect the activity of myocardial cells,the purity was detected by immunocytochemical staining.(2)The surface area of myocardial cells was detected by immunological technique.(3)The expression of ANP/β-MHC was detected by qRT-PCR.(4)The protein content of myocardial cells was detected by BCA protein.2.The mouse myocardial cells were randomly divided into normal group,PE group,solvent group(PE + DMSO)and PE + EGCG group.The four groups were tested as follows:(1)The expression of GATA banding protein 4(GATA4)mRNA and protein was detected by qRT-PCR and western blot.(2)The expression of atrial natriurotic peptide(ANP)and beta-myosin heavy chain(β-MHC)was detected by western blot.(3)The expression levels of HDAC2 protein were detected by colorimetric and western blot.(4)The surface area of myocardial cells was detected by immunological technique.Results:(1)The study found that primary myocardial cells have been cultured and have a purity of more than 90%.The CCK-8 method was used to compare the survival rate of myocardial cells with different concentrations of PE.Compared with the control group,there was no significant difference in the survival rate of myocardial cells when PEconcentration was less than 100 μmol / L(P > 0.05).(2)Compared with the normal group,the expression and activity of HDAC2 protein in PE group and solvent group were increased(P < 0.01),;GATA4 protein,mRNA expression and cardiac hypertrophy markers ANP,β-MHC protein expression were significantly up-regulated(P < 0.01),the difference was statistically significant;myocardial surface area increased(P < 0.01).(3)Immunocytochemical staining results showed that compared with PE group and solvent group,the myocardial surface area was apparently decreased(P < 0.01).(4)Western blot results showed that compared with PE group and solvent group,HDAC2 protein expression and activity were significantly decreased in EGCG group(P <0.01).(5)Western blot results showed that compared with PE group and solvent group,GATA4 protein and mRNA expression were significantly decreased in EGCG group(P < 0.01);The expression of ANP and β-MHC protein in cardiac hypertrophy markers was down-regulated significantly in the same samples(P < 0.01).Conclusions:(1)EGCG can improve cardiac hypertrophy in mice induced by phenylephrine.(2)EGCG can improve cardiac hypertrophy in mice maybe by down-regulating HDAC2 expression.