家兔心臟交感神經(jīng)分泌的在體（in vivo）研究：電化學微電極實時記錄心室區(qū)誘發(fā)交感兒茶酚胺釋放
發(fā)布時間：2019-05-12 19:45
【摘要】： 背景:心臟交感神經(jīng)的激活在心血管疾病的病理生理過程中起著重要的作用,常用的檢測交感神經(jīng)遞質(zhì)釋放的方法,包括微透析技術(shù)和同位素稀釋法等方法不能實時地記錄心臟去甲腎上腺素(NE)的釋放。恒壓安培法(Amperometry)是一種電化學方法,對組織損傷小且具有較高的時間和空間分辨率。本學位論文首先設計出適用于心臟在體(in vivo)記錄的新方法,在家兔心臟上用安培法在體記錄誘發(fā)的NE釋放。目的:通過對原有的碳纖電極(CFE)技術(shù)的改進,創(chuàng)建一種用安培法在體記錄心臟交感神經(jīng)NE釋放的新實驗方法,并研究缺氧和交感興奮誘導的心室NE釋放信號。方法:1.日本大耳白兔心包懸吊法暴露心臟,固定裝置固定心臟。2.我與呂靖、修蕓同學改造了現(xiàn)有的玻璃CFE,使用金絲制成聚乙烯固定式金電極(GFE)和浮動式GFE。在導師的指導下,我設計了創(chuàng)新的浮動電極專用記錄支架,使GFE電極能夠穩(wěn)定地檢測到缺氧或電刺激神經(jīng)誘導的來自位于心室內(nèi)微米區(qū)域內(nèi)交感神經(jīng)末梢釋放的NE信號。3.創(chuàng)建雙通道浮動式GFE同時記錄心臟左右心室NE釋放的方法。結(jié)果:1.體外實驗顯示GFE能夠定量檢測到NE釋放,具有良好的信噪比。2.CFE能夠記錄到短暫窒息誘導的心交感神經(jīng)NE釋放,重復性良好,發(fā)現(xiàn)重復窒息缺氧誘發(fā)心神經(jīng)NE釋放量逐漸減少的現(xiàn)象。3.GFE能夠記錄缺氧或電刺激頸部交感神經(jīng)(SG)和副交感神經(jīng)(VN)誘導的心交感神經(jīng)NE釋放,重復性良好。4.浮動式GFE能夠記錄電刺激SG和VN誘導心交感神經(jīng)末梢NE的釋放,發(fā)現(xiàn)GFE記錄的信號是包含心電和NE的混合信號。5.浮動式GFE雙通道記錄出刺激頸部SG誘導左右心室交感神經(jīng)NE的釋放。結(jié)論:本學位研究過程中設計出了固定式GFE和浮動式GFE兩種實驗記錄方法,兩種方法均能夠有效地記錄心肌NE的釋放。特別是,浮動式GFE的方法簡單、實用、重復性好、對組織損傷小,可以應用于研究生理或病理狀態(tài)下心臟交感神經(jīng)NE釋放信號。應用這項創(chuàng)新的在體記錄技術(shù),我與呂靖、修蕓同學一起第一次在家兔心室上在體實時記錄到誘發(fā)的心臟NE神經(jīng)遞質(zhì)的釋放。
[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.
【學位授予單位】：中國協(xié)和醫(yī)科大學
【學位級別】：博士
【學位授予年份】：2008
【分類號】：R33
本文編號：2475652
[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.
【學位授予單位】：中國協(xié)和醫(yī)科大學
【學位級別】：博士
【學位授予年份】：2008
【分類號】：R33
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