發(fā)布時間：2018-03-03 20:38

  本文選題：心肌細胞肥大　切入點：心臟發(fā)育相關轉錄因子　出處：《重慶醫(yī)科大學》2017年碩士論文　論文類型：學位論文

【摘要】：目的觀察在心肌細胞肥大過程中相關基因時序性表達規(guī)律,與心臟發(fā)育過程中相關規(guī)律進行對比,為心肌細胞肥大機制研究及治療提供線索。方法選擇出生1-3天的昆明乳鼠進行心肌細胞原代培養(yǎng),用異丙腎上腺素(ISO)干預心肌細胞,用CCK-8、細胞計數儀、細胞免疫組化分別檢測心肌細胞存活率、平均直徑及表面積,篩選出構建模型的最佳ISO濃度。用RT-PCR、Western Blot分別檢測相關基因mRNA及蛋白表達水平。結果用不同濃度ISO干預心肌細胞48h后細胞體積在隨濃度濃度增加體積稍增大,到5μmol/L時達峰值(P0.05),后體積稍有減小(P0.05)。不同濃度ISO干預心肌細胞48h后,各組細胞存活率較正常對照組無明顯變化(P0.05),但20μmol/L組細胞存活率明顯低于5μmol/L及10μmol/L組(P0.05)。5μmol/L ISO干預心肌細胞48h細胞表面積較正常對照組明顯增大(P0.05)。5μmol/L ISO干預后,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表達呈先升后降的趨勢,至干預72h GATA4、SRF、α-MHC、β-MHC、P300 mRNA表達仍高于正常(P0.05),而MEF2C降至正常(P0.05);GATA5、TBX5、ANP、BNP mRNA表達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48h后,GATA4、HEY2蛋白表達水平分別升高、降低(P0.05)。結論根據細胞體積、細胞存活率、細胞表面積及肥大基因的表達結果提示心肌細胞肥大模型構建成功。在該心肌細胞肥大模型中,MEF2C的時序表達規(guī)律與心臟發(fā)育過程相似,而GATA4、GATA5、SRF、TBX5的時序性表達規(guī)律與心臟發(fā)育過程不完全一致,提示其在心肌細胞肥大由代償轉為失代償過程中的重要作用。同時GATA4、MEF2C、SRF與乙�；�酶P300的時序性表達規(guī)律相似,提示在心肌細胞肥大過程中其時序性表達可能受P300調控。
[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：1562648