TRP通道药物筛选与通道特性研究

发布时间：2018-01-13 03:30

本文关键词：TRP通道药物筛选与通道特性研究　出处：《中国科学院大学(中国科学院上海药物研究所)》2017年硕士论文　论文类型：学位论文

【摘要】：钙离子对细胞的生理功能有很大的作用,能够促进细胞的生长、增殖,以及凋亡等等。瞬时受体电位离子通道TRP channel多数具有钙离子通透性,共分为七大类,包括TRPA(ANKTM1)、TRPC(canonical)、TRPM(melastatin),TRPML(mucolipin)、TRPP(polycystin)、TRPV(vanilloid)、TRPN(NOMPC)。其中,TRPN(NOMPC)只在非脊椎动物中表达。利用电生理膜片钳技术及相关的分子实验操作手段,本论文在三个方面(筛选针对TRPC通道的小分子调节剂;研究TRPV1通道pore helix区带电荷氨基酸的电生理特性;探讨TRPV4通道在胃癌细胞中可能的作用机制)对TRP通道进行了研究,并得到了相应的实验结果。TRPC通道在许多组织中都有表达,比如肾脏,平滑肌,神经系统等。TRPC通道通过调节钙离子内流的方式来维持细胞的生理功能,其蛋白异常表达会导致许多相关疾病。因此,特异性的TRPC通道小分子调节剂对于研究TRPC通道的功能以及治疗相关疾病都有着十分重要的意义。然而目前针对TRPC通道的特异性小分子激动剂和抑制剂都十分罕见,筛选出特异性的TRPC通道小分子调节剂非常必要。本文第一部分基于对通过高通量筛选发现的一个TRPC6的先导化合物的电生理学鉴定,确定其为TRPC3/6/7通道的特异性激动剂并发现了这三种通道在同等刺激条件下对细胞外钙离子的不同敏感性。TRPV1通道于1997年首先在大鼠背根神经节神经元中被克隆发现,并证明伤害性热刺激和辣椒素所引发的感受性疼痛均是通过TRPV1通道所介导。TRPV1通道的克隆促进了对于感觉神经元中伤害性热刺激和化学刺激传导分子机制的研究。已经有多种关于激动或上调TRPV1通道的机制被详细研究并报导,其中辣椒素,酸,温度和电压都是激动TRPV1通道的主要方式。本文第二部分旨在探讨TRPV1孔道侧的alpha螺旋(Pore Helix)对TRPV1通道门控的作用。发现这一区域中带电荷氨基酸残基对于细胞外酸和电压引起的TRPV1通道开放起着关键作用。研究结果加深了对于TRPV1通道门控机理的了解。钙功能失调在胃癌(Gastric Cancer,GC)细胞中非常常见,也是影响胃癌发生发展的一个主要因素。补钙不论对于健康人还是癌症患者都是一种常见的营养补充方式。其直接影响就是提高胃肠表皮细胞外钙离子的浓度。但是细胞外钙离子浓度的升高对胃癌细胞的生长有无影响,是促进还抑制,目前来讲尚无定论。钙敏感受体(Ca~(2+)-sensitive receptor,CaSR)是表达在细胞膜上感受细胞外钙离子浓度的G蛋白偶联受体,在胃癌细胞中也有表达。本文第三部分旨在探讨胃癌细胞中钙敏感受体的功能以及其如何通过和TRPV4离子通道相互作用实现对钙离子内流的调控来影响细胞增殖。研究结果对治疗胃癌和鉴定新的抗癌的靶点分子都有很大意义。
[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.
【学位授予单位】：中国科学院大学(中国科学院上海药物研究所)
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R96

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