敬钊缨毛蛛粗毒对新生大鼠心室肌细胞电生理特性的影响

发布时间：2018-07-09 20:46

本文选题：敬钊缨毛蛛粗毒 + 新生大鼠心室肌细胞(NRVMs)　；参考：《湖南师范大学》2015年硕士论文

【摘要】：心血管疾病对人们健康的威胁日趋严重与明显,被喻为“三大杀手”之一。大量的研究已经证明了心脏离子通道的电生理特性直接影响着心肌细胞的静息电位以及心脏动作电位的形状和时长。因此,我们通过利用不同的检测方法来试图探讨敬钊缨毛蛛粗毒与心脏离子通道相互作用的电生理特点,从而为挖掘找到潜在的作用靶点或寻找工具分子奠定基础。敬钊缨毛蛛(Chilobrachys jingzhao)是发现于我国海南地区的一种有毒蜘蛛,从该种蜘蛛毒腺中获取的粗毒具有大量的毒素肽物质,就好比一个丰富的毒素肽库。然而,目前关于敬钊粗毒对心脏离子通道影响的研究少之又少。本文,旨在初步探究敬钊粗毒--这个充满潜力的毒素库--中是否也存在有望成为治疗心脏疾病的药用成分,所以首先利用敬钊粗毒来初步探究其对新生大鼠心室肌(neonatal rat ventricular myocytes(NRVMs))相关电生理的影响关系。为此,在这项研究中我利用了全细胞膜片钳技术并通过三个不同的方面开展研究:1,检测敬钊粗毒对新生大鼠心室肌细胞的动作电位的影响;2,检测敬钊粗毒对新生大鼠心室肌细胞的全电流;3,进一步分别检测敬钊粗毒对新生大鼠心室肌细胞上各种电流,即钠电流(INa)、内向整流钾电流(IK1)、快激活延迟整流钾电流(IKr)、瞬时外向钾电流(Ito1)、慢激活延迟整流钾电流(IKs)、钙电流等单独存在时的作用。我们发现,100μg/m L敬钊粗毒对ICaL的增强作用明显,增加幅度为132±4.0%,而对INa抑制了96.7±3.4%,对IKr的峰电流、尾电流以及IKr去极化末部的电流、的抑制分别为54.3±3.2%、60.8±4.3%和35.3±5.3%。另外,该粗毒对IKs,Ito1和IK1电流没有明显的影响,并且这些分别检测的离子电流数据与检测全电流得到的实验结果是吻合的,这表明敬钊粗毒对新生大鼠心室肌离子通道作用具有复杂性,表现出一个多方面的药理作用。此外,通过敬钊粗毒对NRVMs的动作电位有微弱的延长作用并且这种延长作用与动作电位的刺激频率有关联。综上可知,敬钊粗毒对NRVMs上的多种离子通道的电生理特性有影响,具有成为研发治疗心脏疾病药用分子的潜力。
[Abstract]:Cardiovascular disease is becoming more and more serious and obvious to people's health, and it is regarded as one of the "three killers". A large number of studies have proved that the electrophysiological characteristics of cardiac ion channels directly affect the resting potential of cardiac cells and the shape and duration of cardiac action potential. Therefore, we try to explore the electrophysiological characteristics of the interaction between the crude venom of Spider Jingzhao and the ion channel of the heart by using different detection methods, so as to lay a foundation for finding potential targets or searching for tool molecules. Chilobrachys jingzhao is a poisonous spider found in Hainan area of China. The crude toxin obtained from the venomous gland of the spider has a large amount of toxin peptides, which is like a rich toxin peptide library. However, there are few studies on the effect of Jingzhao crude toxin on cardiac ion channels. The purpose of this paper is to preliminarily explore whether there is also a medicinal component in Jingzhao crude toxin, which is a potential toxin pool, that could be used to treat heart disease. Therefore, the effects of Jingzhao crude toxin on (neonatal rat ventricular myocytes (NRVMs in neonatal rat ventricular muscle were studied. To this end, In this study, I made use of whole-cell patch clamp technique and carried out a study on the effect of Jingzhao crude toxin on the action potential of ventricular myocytes in neonatal rats by using three different aspects, I. e., the effects of Jingzhao crude toxin on the heart of newborn rats and the effects of Jingzhao crude toxin on the heart of newborn rats. The total current of ventricular myocytes (VMC) was measured respectively to detect the various currents of Jingzhao crude toxin on ventricular myocytes of newborn rats. Such as sodium current (Ina), inward rectifier potassium current (IK1), fast activated delayed rectifier potassium current (IKr), transient outward potassium current (Ito1), slow activation delayed rectifier potassium current (IKs), calcium current and so on. We found that 100 渭 g / mL Jingzhao crude toxin enhanced ICaL obviously by 132 卤4.0, but inhibited 96.7 卤3.4 of INa, 54.3 卤3.2% of peak current and tail current of IKr and 35.3 卤5.3 卤5.3of the end of depolarization of IKr, respectively. In addition, the crude poison has no obvious effect on the Ito1 and IK1 currents, and these ion current data measured by the two methods are in agreement with the experimental results obtained from the detection of the full current. This indicates that Jingzhao crude toxin has a complex effect on the ion channel in neonatal rat ventricular myocytes, and it shows a variety of pharmacological effects. In addition, the action potential of NRVMs was slightly prolonged by Jingzhao crude toxin, and the prolongation was related to the stimulation frequency of action potential. It can be concluded that Jingzhao crude toxin has an effect on the electrophysiological characteristics of various ion channels on NRVMs and has the potential to be a medicinal molecule for the treatment of heart disease.
【学位授予单位】：湖南师范大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R54

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