本文關(guān)鍵詞：TRP通道藥物篩選與通道特性研究　出處：《中國科學(xué)院大學(xué)(中國科學(xué)院上海藥物研究所)》2017年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： TRPC通道 特異性小分子激動劑 電生理技術(shù) TRPV1通道 酸pH 5.5 capsaicin 電壓依賴性 TRPV4通道 胃癌 鈣敏感受

【摘要】：鈣離子對細胞的生理功能有很大的作用,能夠促進細胞的生長、增殖,以及凋亡等等。瞬時受體電位離子通道TRP channel多數(shù)具有鈣離子通透性,共分為七大類,包括TRPA(ANKTM1)、TRPC(canonical)、TRPM(melastatin),TRPML(mucolipin)、TRPP(polycystin)、TRPV(vanilloid)、TRPN(NOMPC)。其中,TRPN(NOMPC)只在非脊椎動物中表達。利用電生理膜片鉗技術(shù)及相關(guān)的分子實驗操作手段,本論文在三個方面(篩選針對TRPC通道的小分子調(diào)節(jié)劑;研究TRPV1通道pore helix區(qū)帶電荷氨基酸的電生理特性;探討TRPV4通道在胃癌細胞中可能的作用機制)對TRP通道進行了研究,并得到了相應(yīng)的實驗結(jié)果。TRPC通道在許多組織中都有表達,比如腎臟,平滑肌,神經(jīng)系統(tǒng)等。TRPC通道通過調(diào)節(jié)鈣離子內(nèi)流的方式來維持細胞的生理功能,其蛋白異常表達會導(dǎo)致許多相關(guān)疾病。因此,特異性的TRPC通道小分子調(diào)節(jié)劑對于研究TRPC通道的功能以及治療相關(guān)疾病都有著十分重要的意義。然而目前針對TRPC通道的特異性小分子激動劑和抑制劑都十分罕見,篩選出特異性的TRPC通道小分子調(diào)節(jié)劑非常必要。本文第一部分基于對通過高通量篩選發(fā)現(xiàn)的一個TRPC6的先導(dǎo)化合物的電生理學(xué)鑒定,確定其為TRPC3/6/7通道的特異性激動劑并發(fā)現(xiàn)了這三種通道在同等刺激條件下對細胞外鈣離子的不同敏感性。TRPV1通道于1997年首先在大鼠背根神經(jīng)節(jié)神經(jīng)元中被克隆發(fā)現(xiàn),并證明傷害性熱刺激和辣椒素所引發(fā)的感受性疼痛均是通過TRPV1通道所介導(dǎo)。TRPV1通道的克隆促進了對于感覺神經(jīng)元中傷害性熱刺激和化學(xué)刺激傳導(dǎo)分子機制的研究。已經(jīng)有多種關(guān)于激動或上調(diào)TRPV1通道的機制被詳細研究并報導(dǎo),其中辣椒素,酸,溫度和電壓都是激動TRPV1通道的主要方式。本文第二部分旨在探討TRPV1孔道側(cè)的alpha螺旋(Pore Helix)對TRPV1通道門控的作用。發(fā)現(xiàn)這一區(qū)域中帶電荷氨基酸殘基對于細胞外酸和電壓引起的TRPV1通道開放起著關(guān)鍵作用。研究結(jié)果加深了對于TRPV1通道門控機理的了解。鈣功能失調(diào)在胃癌(Gastric Cancer,GC)細胞中非常常見,也是影響胃癌發(fā)生發(fā)展的一個主要因素。補鈣不論對于健康人還是癌癥患者都是一種常見的營養(yǎng)補充方式。其直接影響就是提高胃腸表皮細胞外鈣離子的濃度。但是細胞外鈣離子濃度的升高對胃癌細胞的生長有無影響,是促進還抑制,目前來講尚無定論。鈣敏感受體(Ca~(2+)-sensitive receptor,CaSR)是表達在細胞膜上感受細胞外鈣離子濃度的G蛋白偶聯(lián)受體,在胃癌細胞中也有表達。本文第三部分旨在探討胃癌細胞中鈣敏感受體的功能以及其如何通過和TRPV4離子通道相互作用實現(xiàn)對鈣離子內(nèi)流的調(diào)控來影響細胞增殖。研究結(jié)果對治療胃癌和鑒定新的抗癌的靶點分子都有很大意義。
[Abstract]:Calcium ion has a great effect on the physiological function of cells, which can promote cell growth, proliferation, apoptosis and so on. Most of the transient receptor potential channel TRP channel have calcium permeability. They are divided into seven categories, including TRPAA ANKTM1, TRPC canonical albino, TRPMN melastatin. TRPMLS mucolipinus TRPPN polycystinus, TRPVV vanilloididae, TRPN, NOMPC. among them. TRPN- NOMPCs were expressed only in non-vertebrate animals. Electrophysiological patch clamp technique and related molecular techniques were used. In this paper, there are three aspects: screening of small molecular modulators for TRPC channels; The electrophysiological characteristics of charged amino acids in pore helix region of TRPV1 channel were studied. To explore the possible mechanism of TRPV4 channels in gastric cancer cells, we have studied the TRP channels, and obtained the corresponding results. TRPC channels are expressed in many tissues, such as the kidney. Smooth muscle, nervous system and so on. TRPC channels maintain the physiological function of cells by regulating the influx of calcium ions. The abnormal expression of its protein will lead to many related diseases. The specific TRPC channel small molecule modulator is very important for studying the function of TRPC channel and treating related diseases. However, the specific small molecule agonist for TRPC channel at present is very important. And inhibitors are rare. It is necessary to screen specific TRPC channel small molecular modulators. The first part of this paper is based on the electrophysiological identification of a leading compound of TRPC6 found by high-throughput screening. It was identified as a specific agonist for TRPC3/6/7 channels and the different sensitivities of these three channels to extracellular calcium ions under the same stimulation conditions were found. TRPV1 channels were first found to be large on 1997. Rat dorsal root ganglion neurons were cloned. It was proved that nociceptive heat stimulation and capsaicin induced pain were mediated by the cloning of TRPV1 channel. TRPV1 channel promoted the transmission of nociceptive heat stimulation and chemical stimulation in sensory neurons. Studies on the mechanism of molecular conduction. Many mechanisms of activation or upregulation of TRPV1 channels have been studied and reported in detail. Which capsaicin, acid. Temperature and voltage are the main ways to excite the TRPV1 channel. The second part of this paper is to investigate the alpha helix (more Helix) on the TRPV1 channel side. It is found that the amino acid residues with charge in this region play a key role in the opening of TRPV1 channels induced by extracellular acids and voltages. The results of this study have deepened the effect on TRPV. The mechanism of 1 channel gating. Calcium dysfunction in gastric cancer (. Gastric Cancer. GC) cells are very common. Calcium supplementation is a common nutritional supplement for healthy people and cancer patients. The direct effect of calcium supplementation is to increase the concentration of extracellular calcium in gastrointestinal epidermis. However, the increase of extracellular calcium concentration has an effect on the growth of gastric cancer cells. At present, there is no final conclusion. Calcium sensitive receptor (Ca) -sensitive receptor. CaSRs are G protein-coupled receptors that express extracellular calcium concentration on the cell membrane. The third part is to investigate the function of calcium sensitive receptor in gastric cancer cells and how it can influence the fine flow of calcium ion by interacting with TRPV4 ion channel. Cell proliferation. The results are of great significance for the treatment of gastric cancer and the identification of new anti-cancer target molecules.
【學(xué)位授予單位】：中國科學(xué)院大學(xué)(中國科學(xué)院上海藥物研究所)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R96

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