UT-B基因敲除鼠心脏功能与蛋白组学研究

发布时间：2019-05-12 14:39

【摘要】： 目的：揭示UT-B基因缺失导致心肌肥大的分子机制,从而为疾病的防治提供理论依据和新思路。方法：(1)UT-B基因缺失小鼠的饲养；(2)逆转录PCR鉴定UT-B基因敲除小鼠；(3) Realtime PCR鉴定mRNA表达变化请况；(4)乳鼠心肌细胞原代培养；(5)心脏组织尿素浓度测定；(6)免疫染色和组织学分析；(7)超声心动图分析；(8)血液动力学测定；(9)双向电泳分析；(10)质谱分析；(11)Western blot;(12)统计学分析。结果：(1)UT-B基因敲除导致的尿素在心脏组织中的蓄积；(2)UT-B基因敲除导致心脏功能受损；(3)UT-B基因敲除导致的心肌肥厚和心肌纤维化；(4)ANP、TNNT2在UT-B基因敲除小鼠中动态表达；(5)Ras、Rafl、c-myc基因在UT-B基因敲除的小鼠中信号转到通路中有明显改变。结论：(1)UT-B基因敲除导致的小鼠心脏功能障碍及心肌肥厚是一个随年龄变化而加重的过程；(2)UT-B基因敲除导致小鼠心脏传导阻滞使小鼠心脏中蛋白表达的改变是一个动态变化的过程；(3)UT-B基因敲除小鼠导致的小鼠心脏疾病使Ras-Raf信号传导通路发生改变,使癌基因c-myc表达增高。
[Abstract]:Objective: to reveal the molecular mechanism of myocardial hypertrophy caused by UT-B gene deletion, so as to provide theoretical basis and new ideas for the prevention and treatment of diseases. Methods: (1) UT-B gene deletion mice were fed; (2) UT-B knockout mice were identified by reverse transcription PCR; (3) mRNA expression was detected by) Realtime PCR; (4) primary culture of neonatal rat cardiomyocytes; (5) determination of urea concentration in heart tissue; (6) immunostaining and histological analysis; (7) echocardiographic analysis; (8) hemodynamic determination; (9) two-dimensional electrophoresis analysis; (10) mass spectrometry analysis; (11) Western blot; (12) statistical analysis. Results: (1) the accumulation of urea in heart tissue caused by UT-B gene knockout, (2) the damage of cardiac function caused by UT-B gene knockout, (3) myocardial hypertrophy and myocardial fibrosis caused by UT-B gene knockout, and (3) myocardial hypertrophy and myocardial fibrosis caused by UT-B gene knockout. (4) ANP,TNNT2 was dynamically expressed in UT-B knockout mice, and (5) Ras,Rafl,c-myc gene was significantly changed in UT-B knockout mice. Conclusion: (1) the cardiac dysfunction and myocardial hypertrophy induced by UT-B gene knockout in mice are aggravated with the change of age. (2) the change of protein expression in mouse heart caused by cardiac block induced by UT-B gene knockout is a dynamic process. (3) the heart disease caused by UT-B knockout mice changed the Ras-Raf signal transduction pathway and increased the expression of c-myc.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2010
【分类号】：R363

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