UT-B基因敲除小鼠心脏心肌肥大相关分子表达水平的研究

发布时间：2018-04-28 05:49

本文选题：尿素通道蛋白B + 基因敲除　；参考：《吉林大学》2010年硕士论文

【摘要】： 目的:以UT-B -/-小鼠为研究对象,对其心脏形态学变化、心肌肥大相关分子的表达水平进行系列研究,以探讨心肌肥大的发病机制。方法:16周、52周UT-B -/-小鼠和野生型小鼠随机分为四组。取小鼠心脏拍照、称重,固定包埋行HE染色,观察心脏形态学变化。取心脏蛋白匀浆液,Western blotting、RT-PCR检测UT-B蛋白和mRNA的表达。应用Real-time PCR检测心脏组织心肌肥大相关分子ANP、BNP、α-actin的mRNA表达水平。取血清和心脏组织匀浆液检测尿素含量。心脏组织匀浆液检测NO含量。Western blotting、RT-PCR检测eNOs表达水平。结果: UT-B -/-老龄小鼠(52W)心脏体积增大, UT-B -/-老龄小鼠(52W)心脏系数明显高于同龄野生型小鼠,UT-B -/-小鼠血清和心脏组织中尿素水平明显高于UT-B +/+小鼠,UT-B -/-老龄小鼠(52W)增加更为明显,52周UT-B -/-小鼠心脏ANP、α-actin的表达水平明显增加, UT-B-/-小鼠心脏NO水平降低,该小鼠心脏eNOS的mRNA和蛋白表达水平明显低于同龄野生型小鼠,以56周时更为明显。结论:UT-B基因敲除导致小鼠发生了心肌肥大。其机制可能是UT-B基因敲除导致小鼠血液和心肌组织中尿素含量升高,高于正常水平浓度的尿素影响心肌细胞的电生理功能,使心室肌细胞Na+通道电流和K+通道电流受到抑制,心肌细胞的兴奋性和传导性下降,加上长期高尿素内环境对心肌细胞的应激性负荷,及对内源性NO产生的抑制作用,最后导致小鼠发生了心肌肥大,心肌肥大相关分子ANP、α-actin、SERCA2明显增加。
[Abstract]:Aim: to study the changes of cardiac morphology and the expression level of cardiac hypertrophy related molecules in UT-B-r-mice in order to explore the pathogenesis of myocardial hypertrophy. Methods UT-B-r-r-and wild-type mice were randomly divided into four groups. The mice heart was photographed, weighed and immobilized for HE staining to observe the morphologic changes of the heart. The expression of UT-B and mRNA in heart homogenate was detected by RT-PCR. Real-time PCR was used to detect the expression of mRNA and 伪 -actin in cardiac hypertrophy. Serum and heart homogenate were taken to detect urea content. No content in heart homogenate was detected. RT-PCR was used to detect the expression of eNOs in heart tissue homogenate. Results: the heart volume of UT-B -r-aged mice increased, and the heart coefficient of UT-B -r-aged mice was significantly higher than that of UT-B / R mice of the same age. The levels of urea in serum and heart tissue of mice of the same age were significantly higher than those of UT-B / UT-B / R old mice. The expression of ANP- 伪 -actin in the heart of UT-B-/-mice was significantly increased, and the no level in the heart of UT-B-R-mice was decreased. The expression of mRNA and protein in the heart of the mice was significantly lower than that of the wild type mice of the same age, especially at 56 weeks. Conclusion the knockout of the 1: UT-B gene leads to cardiac hypertrophy in mice. The mechanism may be that knockout of UT-B gene leads to the increase of urea content in blood and myocardial tissue of mice. Urea at higher than normal level affects the electrophysiological function of cardiac myocytes and inhibits the Na and K channel currents in ventricular myocytes. The decrease of excitability and conductivity of cardiomyocytes, the stress load of long-term high urea environment on cardiomyocytes, and the inhibition of endogenous no production resulted in cardiac hypertrophy in mice. ANP- 伪-actinine SERCA2 increased significantly.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R363

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