摘要
目的探讨表没食子儿茶素没食子酸酯(Epigallocatechin gallate,EGCG)通过抑制组蛋白去乙酰化酶(Histone deacetylases,HDACs)减轻苯肾上腺素(Phenylephrine,PE)诱导的小鼠心肌细胞肥大,为临床防治肥厚性心肌病提供新的理论依据。方法酶消化法分离获得小鼠心肌细胞,并将细胞随机分为正常组、PE组、溶媒组和EGCG组。实时荧光定量PCR(qRT-PCR)及Western blot检测心脏核心转录因子GATA结合蛋白4(GATA binding protein 4,GATA4)mRNA及蛋白表达量,Western blot检测心肌肥厚标记物心房钠尿肽(Atrial natriuretic peptide,ANP)、β-心肌肌球蛋白重链(Beta myosin heavy chain,β-MHC)及HDAC2蛋白表达量,比色法检测HDAC2活性,免疫荧光检测心肌细胞表面积。结果与正常组比较,PE组HDAC2蛋白表达量及活性均显著升高(P<0.01),心脏核心转录因子GATA4蛋白及mRNA表达量均显著上调(P<0.01),心肌肥厚标记物ANP、β-MHC蛋白表达量显著上调(P<0.01),心肌细胞表面积明显增大(P<0.01);与PE组比较,EGCG组中HDAC2蛋白表达量及活性均显著降低(P<0.01),心脏核心转录因子GATA4蛋白及mRNA表达量均显著下调(P<0.01),心肌肥厚标记物ANP、β-MHC蛋白表达量明显下调(P<0.01),心肌细胞表面积显著减少(P<0.01)。结论EGCG可能通过调控HDAC2的表达进而改善苯肾上腺素诱导的小鼠心肌细胞肥大。
Objective To explore whether Epigallocatechin gallate(EGCG)attenuated phenylephrine-induced cardiac hypertrophy by inhibiting histone deacetylase(HDAC)in mouse cardiomyocyte,and further provide a new idea for preventing and curing hypertrophic cardiomyopathy.Methods Myocardial cells of mice were isolated by enzymatic digestion and randomly divided into normal group,phenylephrine(PE)group,solvent group(dimethylsulfoxide+PE)and EGCG(EGCG+PE)group.The expression of GATA banding protein 4(GATA4)mRNA and protein was detected by qRT-PCR and Western blot assay,respectively.The expression of atrial natriurotic peptide(ANP)and beta-myosin heavy chain(β-MHC)was detected by Western blot assay.The activity of HDAC2 was assayed by colorimetry.Results The results of Western blot assay and colorimetry showed that the expression and activity of HDAC2 in PE group were increased significantly compared with that of normal group.The mRNA level of GATA4 and the protein level of GATA4,ANP andβ-MHC were apparently increased in the same samples.Moreover,the surface area of myocardial cells of mice in PE group was apparently increased compared with that of normal group.Noteworthily,the expression and activity of HDAC2 protein in EGCG group were decreased significantly compared with that of PE group.The mRNA level of GATA4 and the protein level of GATA4,ANP andβ-MHC were apparently decreased in the same samples.In addition,the surface area of myocardial cells of mice in EGCG group was decreased significantly compared with that of PE group.Conclusion EGCG attenuates cardiomyocyte hypertrophy induced by phenylephrine in mice and the mechanisms might be through regulating the expression of HDAC2.
作者
邓玲
彭波辉
彭昌
Deng Ling;Peng Bohui;Peng Chang(Department of Pediatrics,Affiliated Hospital of Zunyi Medical University,Zunyi Guizhou 563099,China)
出处
《遵义医科大学学报》
2020年第2期155-159,共5页
Journal of Zunyi Medical University
基金
国家自然科学基金资助项目(NO:81560040)
遵义医学院博士启动基金资助项目(NO:院字[2015]4)
遵义医学院与科技学院大学生创新训练项目(NO:遵医科院[2015]3108)。
