尼古丁对干细胞心肌分化的影响研究

发布时间：2018-05-27 23:15

本文选题：尼古丁 + 小鼠胚胎干细胞　；参考：《浙江大学》2010年硕士论文

【摘要】：烟碱(Nicotine),俗名尼古丁,是一种存在于茄科植物中的生物碱,也是烟草的重要成分。当尼古丁进入人体后,会产生很多作用：如四肢末梢血管收缩、呼吸变快、心跳加快、血压上升、精神状况发生改变(例如变得情绪稳定或精神兴奋),并促进血小板凝集,是造成心脏血管堵塞、高血压、中风等心脏血管性疾病的主要帮凶。尼古丁会使人上瘾或者产生依赖性,重复使用尼古丁会增加心脏搏动速率、升高血压并降低食欲。另外,有研究表明,尼古丁可直接影响精子生成,阻碍精子的发生和成熟。胚胎干细胞(embryonic stem cells, ESCs)是从哺乳动物囊胚时期的内细胞团(InnerCellMass)的分离出来的,能够在体外保持未分化状态,并且能够进行高度增殖的多潜能细胞,它具有体外培养无限增殖、自我更新和多向分化等特性。无论在体外还是体内,胚胎干细胞都能被诱导分化为几乎所有的细胞类型,它可以发展为胚体后的生物系统,用于模拟体内细胞与组织间复杂的相互作用,在药物研究领域具有广泛的用途。胚胎干细胞最重要的应用是分化为各种类型的细胞,用于细胞疗法,为细胞移植提供无免疫原性的材料,任何涉及丧失正常细胞的疾病,都可以通过移植由胚胎干细胞分化而来的特异性组织细胞进行治疗。例如用胰岛细胞治疗糖尿病,用神经细胞治疗神经退行性疾病(帕金森病、亨氏舞蹈症等),用心肌细胞修复坏死的心肌等。本文通过经典的心肌细胞诱导方法,首先用小鼠胚胎干细胞制备得到拟胚体(embryoid bodies, EB s),再通过诱导分化获得心肌细胞。设立常规培养的阴性对照,正常心肌诱导的阳性对照以及加入不同浓度尼古丁的实验组,经过15天的诱导后,RT-PCR结果显示,与阳性对照组相比,尼古丁促进了mES干性基因Oct-4,Rex-1的表达,抑制了心肌分化标志基因α-actin, desmin, cTnI表达,提示尼古丁促进mES细胞增殖的同时对心肌分化有一定的抑制作用。临床研究表明,女性在怀孕期间吸烟,会导致婴儿心室间隔缺损和右侧阻塞性缺陷；接受过心脏移植的病人即使少量吸烟,都会增加其发病率和死亡率的危险性。本实验为临床治疗方案的确定以及使用尼古丁戒烟产品存在的潜在风险阐明了理论基础
[Abstract]:Nicotine, commonly known as nicotine, is an alkaloid found in plants of the family Solanaceae, and is also an important component of tobacco. When nicotine enters the body, it has a number of effects: contracting blood vessels at the extremities, breathing faster, beating faster, rising blood pressure, changing mental conditions (e.g. becoming emotionally stable or psychologically excited, and promoting platelet agglutination). It is a major accomplice in cardiovascular diseases such as heart blockage, high blood pressure, stroke and so on. Nicotine can be addictive or dependent. Repeated use of nicotine increases heart rate, raises blood pressure and reduces appetite. In addition, studies have shown that nicotine directly affects spermatogenesis and hinders spermatogenesis and maturation. Embryonic stem cells, ESCs) is isolated from the inner cell mass of mammalian blastocyst. It can maintain undifferentiated and highly proliferative multipotential cells in vitro, and it has unlimited proliferation in vitro. Self-renewal and multi-differentiation. In vitro and in vivo, embryonic stem cells can be induced to differentiate into almost all cell types, which can be developed into post-embryonic biological systems that mimic complex interactions between cells and tissues in vivo. It has a wide range of applications in the field of drug research. The most important application of embryonic stem cells is to differentiate into various types of cells, to be used in cell therapy, to provide non-immunogenic materials for cell transplantation, any disease involving the loss of normal cells. Can be treated by transplanting specific histocytes derived from embryonic stem cells. For example, islet cells are used to treat diabetes, nerve cells to treat neurodegenerative diseases (Parkinson's disease, Henderson's chorea, etc.), cardiomyocytes to repair necrotic myocardium, and so on. In this paper, embryoid bodieses, EB sr. were prepared from mouse embryonic stem cells by classical cardiomyocyte induction method, and then cardiomyocytes were obtained by inducing differentiation. After 15 days of induction, the negative control group, the positive control group induced by normal myocardium and the experimental group with different concentrations of nicotine were established. The results of RT-PCR showed that compared with the positive control group, Nicotine promoted the expression of Oct-4N Rex-1, inhibited the expression of 伪 -actinin, desmin and cTnI, suggesting that nicotine could inhibit the proliferation of mES cells and inhibit myocardial differentiation. Clinical studies have shown that smoking during pregnancy can lead to ventricular septal defect and right obstructive defect in infants, and that even a small amount of smoking in patients undergoing heart transplantation increases the risk of morbidity and mortality. This experiment clarifies the theoretical basis for the determination of clinical treatment plan and the potential risk of nicotine smoking cessation products.
【学位授予单位】：浙江大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R329

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