心肌细胞肥大过程中心脏发育相关转录因子及下游心脏肥大标志基因的时序性表达规律研究  

The temporal expression patterns of the transcription factors and target genes in cardiomyocyte hypertrophy

作　　者：罗漫[1] 吴晓云[1] 郑敏[1] 刘玲娟[1] 田杰[1] 

机构地区：[1]重庆医科大学附属儿童医院心血管内科与小儿心血管病研究室、儿童发育疾病研究省部共建教育部重点实验室、重庆市儿童发育重大疾病诊治与预防国际科技合作基地、儿科学重庆市重点实验室,重庆400014

出　　处：《解放军医学杂志》2017年第5期394-401,共8页Medical Journal of Chinese People's Liberation Army

基　　金：国家自然科学基金(81270234,81570218);重庆市社会民生科技创新专项项目(cstc2015shmszx120043)~~

摘　　要：目的观察在心肌细胞肥大过程中心脏发育相关转录因子及下游心脏肥大标志基因时序性表达规律,为心肌肥大机制研究提供线索。方法用异丙肾上腺素(ISO)干预心肌细胞,用CCK-8、细胞计数仪、免疫组化分别检测细胞存活率、细胞直径及表面积,用RT-PCR、Western blotting分别检测相关基因mRNA及蛋白表达水平。结果用ISO成功构建心肌细胞肥大模型。ISO干预后,GATA结合蛋白4(GATA4)、肌细胞增强因子2C(MEF2C)、GATA5、脑钠肽(BNP)、心房利钠因子(ANP)于24h开始升高,P300、α-肌球蛋白重链(α-MHC)、T-框蛋白5(TBX5)于12h开始升高,血清应答因子(SRF)、β-MHC mRNA于6h开始升高(P<0.05);其中GATA4、α-MHC、β-MHC、SRF、P300 mRNA表达呈先升后降的趋势,至干预72h GATA4、SRF、α-MHC、β-MHC、P300 mRNA表达仍高于正常(P<0.05),而MEF2C降至正常(P>0.05);GATA5、TBX5、ANP、BNP mRNA表达与对照组相比持续升高(P<0.05),而NKX2.5 mRNA(P>0.05)无明显变化。ISO干预48h后,GATA4蛋白表达水平升高,HEY2蛋白表达水平降低(P<0.05)。结论在心肌细胞肥大过程中,MEF2C的时序表达规律与心脏发育过程相似,而GATA4、GATA5、SRF、TBX5的时序性表达规律与心脏发育过程不完全一致,提示其在心肌细胞肥大由代偿转为失代偿过程中的重要作用。同时GATA4、MEF2C、SRF与乙酰化酶P300的时序性表达规律相似,提示在心肌细胞肥大过程中其时序性表达可能受P300调控。Objective To investigate the temporal expression patterns of the related transcription factors and target genes in cardiomyocyte hypertrophy, and provide valuable clues for further researches on cardiomyocyte hypertrophy. Methods Isoproterenol （ISO） was used to induce ventricular myocytes hypertrophy in neonatal mice. The survival rate of cardiomyocyte was detected by CCK-8, and the average diameters and surface areas of cells were measured by computer photograph analysis system. The mRNA and protein expression levels of related genes were respectively measured by RT-PCR and Western blotting. Results The model of cardiomyocyte hypertrophy stimulated by ISO was constructed successfully. The expression levels of GATA4, MEF2C, GATAS, BNP and ANP increased 24 hours after ISO treatment, the expression levels of P300, α-MHC and TBX5 increased 12 hours after 1SO treatment, and of SRF and β-MHC mRNA increased 6 hours after ISO treatment （P〈0.05）. The expression levels of GATA4, α-MHC, β-MHC, SRF and P300 mRNA increased firstly, and then decreased in cardiomyocytes induced by ISO. The expression levels of GATA4, SRF, α-MHC, β-MHC and P300 mRNA were still higher than normal （P〈0.05）, but of MEF2C decreased to normal （P〉0.05） 72, hours after ISO treatment. The expression levels continuously elevated of GATA5, TBX5, ANP and BNP mRNA than that of controls （P〈0.05）, while no fluctuation was found in NKX2.5 mRNA expression （P〉0.05）. The expression of GATA4 protein increased, while of HEY2 protein decreased 48 hours after ISO treatment （P〈0.05）. Conclusions In hypertrophic cardiomyocytes, the expression pattern of MEF2C is similar to, but the patterns of GATAS, GATA4, TBX5 and SRF are different with that in the development of heart, implying these genes are important during the process from compensatory stage to decompensation stage. The expression patterns of GATA4, MEF2C and SRF are similar to that of acetylase P300, implying the temporal expressions could be regulated by P300 in

关 键 词：心肌细胞肥大 心脏发育相关转录因子 心脏肥大标志基因 时序性表达 

分 类 号：R322.11[医药卫生—人体解剖和组织胚胎学]