【作者】 杨红；

【导师】 王克玉；

【作者基本信息】 山东大学 ， 皮肤病与性病学， 2006， 硕士

【摘要】 白癜风是一种常见的色素脱失性皮肤病，临床表现为局部或泛发性色素脱失斑。本病可发生于任何年龄，在我国人群中发病率约为0.1％-2.0％。白癜风多发于暴露部位，常给患者带来巨大的心理压力。白癜风的病因及发病机制尚未完全阐明，一般认为是具有遗传素质的个体，在内源性和外源性刺激的作用下导致的免疫、代谢、内分泌及神经精神功能的紊乱。目前病因主要有以下几个方面的假设：自身免疫学说；神经与精神化学学说；遗传学说；黑素细胞自身破坏学说和微量元素缺乏等学说。 近几年来随着自由基在医学上的深入研究，发现在白癜风患者的表皮内出现过氧化氢的过量聚集，过氧化氢是一种较强的氧化剂，它可以与金属离子或氧离子发生反应，产生羟自由基。羟自由基可以引起脂质过氧化反应，损伤细胞。表皮的氧化应激状态可能降低了某些酶的活性，造成黑素细胞的损伤，导致色素脱失。 本试验以过氧化氢作为外源性氧化剂，通过向体外培养的黑素细胞中加入过氧化氢，使黑素细胞处于氧化应激的状态，观察细胞在形态超微结构及功能上的改变，探讨氧化应激与白癜风发病机制的关系。本研究旨在探讨氧化应激对黑素细胞的损伤及其与白癜风发病的关系，为白癜风的预防和治疗提供实验依据。 第一部分 黑素细胞的体外培养及生物学鉴定 目的 为应用培养的黑素细胞研究白癜风的发病机制，对正常人表皮的黑素细胞进行体外原代及传代培养，并对黑素细胞的生物学特性进行鉴定。 方法 采用MCDB254培养基对正常人表皮黑素细胞进行体外培养，采用HMB-45单克隆抗体免疫组化法对培养细胞进行鉴定，透射电镜观察细胞的超微结构，确定培养细胞为黑素细胞且具有正常的生物学特征更多还原

【Abstract】 Vitiligo is a depigmenting disease of the skin ,characterized by destruction of MC in lesions of vitiligo , resulting in patches of depigmentation. In China, the incidence of vitiligo in patients is about 1% to 2%. It can condition the behavior of people suffering from vitiligo and can deeply affect their personal relationships. The etiology is still unknown and pathogenesis has not been completely clarified so far, various hypotheses have been proposed, they include the genetic hypotheses, the autoimmune hypotheses , the autocytotoxic hypotheses, the neural hypotheses, and the lack of microelement hypotheses.In recent years , Increase levels of H₂O₂ were demonstrated in the epidermis of both lesional and nonlesional skin of vitiligo patients, H₂O₂ levels increase in response to oxidative stress. An imbalance of antioxidants was found in the epidermis, correlated to an increased reactive oxygen species（ROS） production. ROS can interact with macrom olecules such as proteins, membrance lipids and nucleic acids, and fina lly cause cells damaged.In this study , Normal melanocytes were cultured with H₂O₂ in different dosages（0-0. 5mmol/L） and different time（0—60minute）, the appearance and function of the cells were analysed .The aim of this study was to evaluate the effect of H₂O₂ on the melanocyte in vitro cultured and the relationship between oxidative stress and vitiligo. Part One Culture and biological characteristics of melanocytes Objective To indentify the biological characteristics of MC cultured in更多还原