家兔心脏交感神经分泌的在体（in vivo）研究：电化学微电极实时记录心室区诱发交感儿茶酚胺释放

发布时间：2019-05-12 19:45

【摘要】： 背景:心脏交感神经的激活在心血管疾病的病理生理过程中起着重要的作用,常用的检测交感神经递质释放的方法,包括微透析技术和同位素稀释法等方法不能实时地记录心脏去甲肾上腺素(NE)的释放。恒压安培法(Amperometry)是一种电化学方法,对组织损伤小且具有较高的时间和空间分辨率。本学位论文首先设计出适用于心脏在体(in vivo)记录的新方法,在家兔心脏上用安培法在体记录诱发的NE释放。目的:通过对原有的碳纤电极(CFE)技术的改进,创建一种用安培法在体记录心脏交感神经NE释放的新实验方法,并研究缺氧和交感兴奋诱导的心室NE释放信号。方法:1.日本大耳白兔心包悬吊法暴露心脏,固定装置固定心脏。2.我与吕靖、修芸同学改造了现有的玻璃CFE,使用金丝制成聚乙烯固定式金电极(GFE)和浮动式GFE。在导师的指导下,我设计了创新的浮动电极专用记录支架,使GFE电极能够稳定地检测到缺氧或电刺激神经诱导的来自位于心室内微米区域内交感神经末梢释放的NE信号。3.创建双通道浮动式GFE同时记录心脏左右心室NE释放的方法。结果:1.体外实验显示GFE能够定量检测到NE释放,具有良好的信噪比。2.CFE能够记录到短暂窒息诱导的心交感神经NE释放,重复性良好,发现重复窒息缺氧诱发心神经NE释放量逐渐减少的现象。3.GFE能够记录缺氧或电刺激颈部交感神经(SG)和副交感神经(VN)诱导的心交感神经NE释放,重复性良好。4.浮动式GFE能够记录电刺激SG和VN诱导心交感神经末梢NE的释放,发现GFE记录的信号是包含心电和NE的混合信号。5.浮动式GFE双通道记录出刺激颈部SG诱导左右心室交感神经NE的释放。结论:本学位研究过程中设计出了固定式GFE和浮动式GFE两种实验记录方法,两种方法均能够有效地记录心肌NE的释放。特别是,浮动式GFE的方法简单、实用、重复性好、对组织损伤小,可以应用于研究生理或病理状态下心脏交感神经NE释放信号。应用这项创新的在体记录技术,我与吕靖、修芸同学一起第一次在家兔心室上在体实时记录到诱发的心脏NE神经递质的释放。
[Abstract]:Background: the activation of cardiac sympathetic nerve plays an important role in the pathophysiological process of cardiovascular disease. The release of norepinephrine (NE) in heart can not be recorded in real time by microdialysis and isotope dilution. Constant pressure amperometric method (Amperometry) is an electrochemical method with small damage to tissue and high temporal and spatial resolution. In this thesis, a new method for in vivo (in vivo) recording of heart was designed, and the release of NE induced by amperometric recording in rabbit heart was recorded in vivo. Aim: to establish a new experimental method for recording cardiac sympathetic NE release in vivo by improving the original carbon fiber electrode (CFE) technique, and to study the ventricular NE release signal induced by hypoxia and sympathetic stimulation. Methods: 1. The pericardial suspension of Japanese white rabbits exposed the heart and fixed the heart. 2. Lu Jing and I modified the existing glass CFE, using gold wire to make polyethylene fixed gold electrode (GFE) and floating GFE. Under the guidance of my mentor, I designed an innovative floating electrode recording stent to enable GFE electrodes to stably detect NE signals from sympathetic nerve endings in the micron region of the ventricle induced by hypoxia or electrical stimulation. 3. A two-channel floating GFE was created to record the NE release from the left and right ventricle of the heart at the same time. Result: 1. In vitro experiments showed that GFE could quantitatively detect the release of NE and had a good signal-to-noise ratio. 2.CFE could record the release of NE from cardiac sympathetic nerve induced by transient asphyxia, and the reproducibility was good. It was found that repeated asphyxia hypoxia induced the gradual decrease of cardiac nerve NE release. 3. Gfe could record the release of cardiac sympathetic nerve NE induced by hypoxia or electrical stimulation of cervical sympathetic nerve (SG) and parasympathetic nerve (VN), and the reproducibility was good. Floating GFE could record the release of NE from cardiac sympathetic nerve endings induced by electrical stimulation of SG and VN. It was found that the signal recorded by GFE was a mixed signal containing ECG and NE. 5. Floating GFE dual channels recorded the release of NE from left and right ventricular sympathetic nerves induced by cervical SG stimulation. Conclusion: two experimental recording methods, fixed GFE and floating GFE, have been designed in the course of this degree study, both of which can effectively record the release of myocardial NE. In particular, the method of floating GFE is simple, practical, reproducible and has little tissue damage. It can be used to study the NE release signal of cardiac sympathetic nerve in physiological or pathological state. Using this innovative in-vivo recording technique, together with Lu Jing and Xiuyun, I recorded the release of cardiac NE neurotransmitters in real time on the ventricle of rabbits for the first time.
【学位授予单位】：中国协和医科大学
【学位级别】：博士
【学位授予年份】：2008
【分类号】：R33

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