發(fā)布時(shí)間：2018-03-03 20:38

  本文選題：心肌細(xì)胞肥大　切入點(diǎn)：心臟發(fā)育相關(guān)轉(zhuǎn)錄因子　出處：《重慶醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的觀察在心肌細(xì)胞肥大過程中相關(guān)基因時(shí)序性表達(dá)規(guī)律,與心臟發(fā)育過程中相關(guān)規(guī)律進(jìn)行對比,為心肌細(xì)胞肥大機(jī)制研究及治療提供線索。方法選擇出生1-3天的昆明乳鼠進(jìn)行心肌細(xì)胞原代培養(yǎng),用異丙腎上腺素(ISO)干預(yù)心肌細(xì)胞,用CCK-8、細(xì)胞計(jì)數(shù)儀、細(xì)胞免疫組化分別檢測心肌細(xì)胞存活率、平均直徑及表面積,篩選出構(gòu)建模型的最佳ISO濃度。用RT-PCR、Western Blot分別檢測相關(guān)基因mRNA及蛋白表達(dá)水平。結(jié)果用不同濃度ISO干預(yù)心肌細(xì)胞48h后細(xì)胞體積在隨濃度濃度增加體積稍增大,到5μmol/L時(shí)達(dá)峰值(P0.05),后體積稍有減小(P0.05)。不同濃度ISO干預(yù)心肌細(xì)胞48h后,各組細(xì)胞存活率較正常對照組無明顯變化(P0.05),但20μmol/L組細(xì)胞存活率明顯低于5μmol/L及10μmol/L組(P0.05)。5μmol/L ISO干預(yù)心肌細(xì)胞48h細(xì)胞表面積較正常對照組明顯增大(P0.05)。5μmol/L ISO干預(yù)后,GATA4、MEF2C、GATA5、BNP、ANP于24h,P300、α-MHC、TBX5于12h,SRF、β-MHC mRNA于6h開始升高(P0.05);其中GATA4、SRF、MEF2C、α-MHC、β-MHC、P300 mRNA表達(dá)呈先升后降的趨勢,至干預(yù)72h GATA4、SRF、α-MHC、β-MHC、P300 mRNA表達(dá)仍高于正常(P0.05),而MEF2C降至正常(P0.05);GATA5、TBX5、ANP、BNP mRNA表達(dá)與正常對照組相比持續(xù)升高(P0.05),而NKX2.5 mRNA無明顯變化(P0.05)。ISO干預(yù)48h后,GATA4、HEY2蛋白表達(dá)水平分別升高、降低(P0.05)。結(jié)論根據(jù)細(xì)胞體積、細(xì)胞存活率、細(xì)胞表面積及肥大基因的表達(dá)結(jié)果提示心肌細(xì)胞肥大模型構(gòu)建成功。在該心肌細(xì)胞肥大模型中,MEF2C的時(shí)序表達(dá)規(guī)律與心臟發(fā)育過程相似,而GATA4、GATA5、SRF、TBX5的時(shí)序性表達(dá)規(guī)律與心臟發(fā)育過程不完全一致,提示其在心肌細(xì)胞肥大由代償轉(zhuǎn)為失代償過程中的重要作用。同時(shí)GATA4、MEF2C、SRF與乙�；�酶P300的時(shí)序性表達(dá)規(guī)律相似,提示在心肌細(xì)胞肥大過程中其時(shí)序性表達(dá)可能受P300調(diào)控。
[Abstract]:Objective To observe the effect in the process of myocardial hypertrophy associated gene sequential expression rules and related rules and heart development comparison, provide clues for the study of mechanism of myocardial cell hypertrophy and treatment methods. Kunming rat was born 1-3 days of myocardial cells cultured with isoproterenol (ISO) on myocardial cells with CCK-8, cell count, cell immunohistochemistry were used to detect the myocardial cell survival rate, average diameter and surface area, screening the best concentration of ISO to construct the model. Using RT-PCR, Western Blot were used to detect related gene mRNA and protein expression level. The results with different concentrations of ISO on myocardial cells after 48h cells with the concentration increase in volume the volume increased slightly to 5, reached the peak at mol/L (P0.05), the volume is slightly decreased (P0.05). Different concentrations of ISO on myocardial cell 48h, cell survival rate compared with the normal control group. 鏃犳槑鏄懼彉鍖，

本文編號(hào)：1562648