本文選題：HCN通道 + KT5720　； 參考：《華中科技大學(xué)》2013年博士論文

【摘要】：細(xì)胞是生命活動(dòng)的基本單位。一切有機(jī)體都是由細(xì)胞構(gòu)成的，它是有機(jī)體生長(zhǎng)與發(fā)育的基礎(chǔ)，也是遺傳的基本單位，因此，沒(méi)有細(xì)胞就沒(méi)有完整的生命。所有的動(dòng)物細(xì)胞都由一層薄膜所包圍，這就是細(xì)胞膜(plasma membrane)，細(xì)胞膜的作用是保持細(xì)胞內(nèi)物質(zhì)成分的穩(wěn)定。但是，，細(xì)胞膜并不是完全封閉的，細(xì)胞需要跟外界交換物質(zhì)和能量，細(xì)胞膜上的離子通道就是具備這種功能的蛋白質(zhì)，離子通道可以分為配體門(mén)控通道、電壓門(mén)控通道、胞內(nèi)信號(hào)門(mén)控通道和機(jī)械敏感通道四種類(lèi)型。 超極化激活的環(huán)核苷酸門(mén)控(hyperpolarization-activated cyclic nucleotide-gated，HCN)通道(起搏通道)是一種能被電壓和環(huán)磷酸腺苷(cAMP)調(diào)控的、無(wú)選擇性陽(yáng)離子通道，它介導(dǎo)的電流是Ih。Ih廣泛的參與了對(duì)一系列生理活動(dòng)的調(diào)控，包括心臟和神經(jīng)元節(jié)律，某些可興奮細(xì)胞的靜息電位，神經(jīng)信號(hào)的傳遞以及信號(hào)在樹(shù)突中的整合等。Ih受自主神經(jīng)系統(tǒng)和神經(jīng)內(nèi)分泌系統(tǒng)的調(diào)節(jié)以及某些病理狀態(tài)的影響，Ih異常會(huì)導(dǎo)致心臟及精神方面的多種疾病。因此，研究Ih有重要的生理學(xué)意義。 本文著重應(yīng)用電生理膜片鉗、全細(xì)胞鈣成像、神經(jīng)疼痛模型等技術(shù)對(duì)HCN通道的調(diào)控機(jī)制、在神經(jīng)疼痛中的作用及其藥理學(xué)特性進(jìn)行了研究和探討。 本文主要研究結(jié)果和結(jié)論如下： （1）PKA信號(hào)通路在調(diào)控HCN通道活性中的作用。PKA廣泛分布于細(xì)胞體內(nèi)，參與了對(duì)一些列重要生理活動(dòng)的調(diào)控。然而，抑制PKA對(duì)HCN通道活性有何影響，目前仍不清楚。為了弄清這一問(wèn)題，我們選擇了PKA的選擇性抑制劑KT5720。在新鮮分離的大鼠DRG神經(jīng)元上，綜合運(yùn)用全細(xì)胞電壓鉗、電流鉗、單通道記錄以及鈣成像技術(shù)。揭示了PKA信號(hào)通路在調(diào)控HCN通道活性中發(fā)揮了極為重要的作用，KT5720抑制HCN通道活性，使其電流密度減小，半數(shù)開(kāi)放電壓向超極化方向移動(dòng)，并使其通道開(kāi)放的時(shí)間常數(shù)增加。此外，KT5720抑制PKA使DRG神經(jīng)元上的動(dòng)作電位減小、胞內(nèi)Ca2+濃度降低，從而抑制DRG神經(jīng)元的興奮性。 （2）HCN通道調(diào)控神經(jīng)疼痛。神經(jīng)疼痛通常由神經(jīng)損傷引發(fā)，因其具有發(fā)病率高、難治愈等特點(diǎn)，已成為當(dāng)今醫(yī)學(xué)界亟待解決的主要問(wèn)題之一。另一方面，DRG神經(jīng)元是感覺(jué)傳入的第一級(jí)神經(jīng)元，在疼痛信號(hào)的傳導(dǎo)過(guò)程中發(fā)揮著不可或缺的重要作用。因此，我們選擇DRG神經(jīng)元為研究對(duì)象，利用大鼠幸免神經(jīng)損傷模型。綜合運(yùn)用多種電生理膜片鉗實(shí)驗(yàn)方法，研究和探討了HCN通道在調(diào)控神經(jīng)疼痛中的機(jī)制，揭示了HCN通道是觸發(fā)和維持神經(jīng)疼痛的關(guān)鍵因素。在幸免神經(jīng)損傷誘導(dǎo)的疼痛狀態(tài)下，DRG神經(jīng)元上的HCN通道活性顯著增加，異位放電頻率變快，且DRG形態(tài)發(fā)生變化。因此，HCN通道是有效治療神經(jīng)疼痛的潛在靶點(diǎn)。 （3）麻黃堿對(duì)HCN通道的影響。麻黃堿作為一種腎上腺受體激動(dòng)藥，具有加強(qiáng)心收縮力、增加心輸出量、加快心率、升高血壓等作用。為了弄清麻黃堿的這些作用是否是通過(guò)HCN通道來(lái)實(shí)現(xiàn)的以及它對(duì)HCN通道電流有什么影響。我們采用傳統(tǒng)的全細(xì)胞膜片鉗實(shí)驗(yàn)方法，在新生的大鼠海馬神經(jīng)元和HEK細(xì)胞上記錄到HCN通道電流Ih。實(shí)驗(yàn)結(jié)果顯示，麻黃堿對(duì)Ih有抑制作用。經(jīng)分析，我們認(rèn)為HCN通道可能具備某種反饋及保護(hù)功能，即當(dāng)血壓和心率上升時(shí)，HCN通道能部分阻滯這種上升趨勢(shì)，從而保護(hù)機(jī)體健康和生命安全。
[Abstract]:Cell is the basic unit of life activity . All organism is composed of cells , it is the base of organism growth and development , it is the basic unit of inheritance , therefore , there is no cell without complete life . All animal cells are surrounded by a layer of membrane .

The hyperpolarization activated cyclic nucleotide - gated ( HCN ) channel is a non - selective cation channel which can be regulated by voltage and cyclic adenosine monophosphate ( cAMP ) . Its current is Ih . Ih is extensively involved in regulation of a series of physiological activities , including heart and neuronal rhythm , resting potential of certain excited cells , transmission of nerve signals , and integration of signals in the dendritic cells . Ih is influenced by autonomic nervous system and neuroendocrine system and the integration of signals in the dendritic cells . Ih can lead to various diseases in the heart and spirit . Therefore , it is important to study Ih .

This paper focuses on the mechanism of HCN channel regulation by using the techniques of electro - physiological patch clamp , whole - cell calcium imaging and neural pain model , and studies and discusses the role and pharmacology of HCN channel .

The main findings and conclusions are as follows :

( 1 ) In order to clarify this problem , we have selected selective inhibitor KT 5720 . In order to understand this problem , we have chosen selective inhibitor of protein kinase , KT 5720 . In order to understand this problem , we have chosen selective inhibitor of the protein of the activity of HCN channel . In order to understand this problem , we have chosen the selective inhibitor of the protein kinase , KT 5720 . In order to find out the problem , we have chosen the selective inhibitor of protein kinase , KT 5720 . In order to find out the problem , we have disclosed the activity of the channel in the freshly isolated rat DRG neurons , and the half - open voltage is decreased , and the time constant of the channel opening is increased . In addition , the inhibition of the activity of the pathway of the channel is reduced , and the concentration of Ca 2 + in the cells is decreased , so that the excitatory of the DRG neurons is inhibited .

In this paper , we select DRG neurons as the research objects and play an important role in the conduction of pain signals .

( 3 ) The effect of ephedrine on HCN channel was studied . The effect of ephedrine on HCN channel was studied .

【學(xué)位授予單位】：華中科技大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2013
【分類(lèi)號(hào)】：R318.04

【參考文獻(xiàn)】

相關(guān)期刊論文 前1條

1 ;Involvement of hyperpolarization-activated,cyclic nucleotide-gated cation channels in dorsal root ganglion in neuropathic pain[J];生理學(xué)報(bào);2008年05期

本文編號(hào)：1777274