重组人Dickkopf1蛋白的克

发布时间：2018-02-27 21:03

本文关键词： dickkopf1 Dkk1蛋白黑素细胞黑素酪氨酸酶　出处：《第四军医大学》2006年硕士论文　论文类型：学位论文

【摘要】：黑素是皮肤成为褐色的主要色素。黑素是酪氨酸在酪氨酸酶作用下产生的，在黑素细胞的黑素小体中生成，黑素小体从一个黑素细胞被转运至一群角质形成细胞。真、表皮内黑素细胞增生及黑素生成功能活跃是色素增多性疾病的一个主要原因。色素增多性疾病在皮肤科疾病中非常常见，最常见的是雀斑、黄褐斑及炎症后色素沉着，，发生在面部、手背等暴露部位显著者严重影响患者的容貌，影响患者的生活质量。根据色素增多性疾病的发病机制，防治措施主要从抑制黑素细胞增殖及活性、防止黑素生成及加速黑素降解等方面入手。目前针对色素增多性疾病防治的各种方法的疗效、复发问题以及可能出现的不良反应，一直是皮肤科临床医师所面临的棘手问题。人掌跖部位皮肤的黑素含量比非掌跖部位低。近来研究发现，掌跖部位皮肤黑素细胞的密度是非掌跖部位皮肤的1／5，黑素小体、黑素相关蛋白含量也相应的显著降低，其原因为掌跖部位皮肤中的成纤维细胞分泌的Dickkopf1蛋白能够通过抑制黑素细胞Wnt信号转导通路的活化进而抑制黑素细胞的增殖、发育、分化及黑素生成能力。本课题通过反转录PCR自人掌跖部位皮肤成纤维细胞中获得了编码Dickkop1的32—266位氨基酸的基因，克隆入PQE30质粒，构建了PQE30-dkk1原核表达质粒，经双酶切鉴定及序列分析证实所获序列完全正
[Abstract]:Melanin is the primary brown pigment of the skin. Melanin is produced by tyrosine acting on tyrosinase, formed in the melanoma body of melanocytes, which is transported from a melanocyte to a group of keratinocytes. Proliferation of melanocytes and active melanogenesis in the epidermis are one of the main causes of hyperpigmentation. Hyperpigmentation is very common in dermatology, and the most common diseases are freckles, chloasma and postinflammatory pigmentation. Those with prominent exposure sites such as the face and the back of the hand seriously affect the appearance of the patient and the quality of life of the patient. According to the pathogenesis of pigmentation disease, the prevention and treatment measures mainly include inhibiting the proliferation and activity of melanocytes. Prevention of melanogenesis and acceleration of melanin degradation, etc. At present, the curative effect, recurrence and possible adverse reactions of various methods for the prevention and treatment of hyperpigmented diseases have been the thorny problems faced by dermatologists. The melanin content of human palmoplantar skin is lower than that of non-palmoplantar skin. Recently, it has been found that the density of melanocytes in palmar plantar skin is 1 / 5 of that in non-palmar metatarsal skin, and melanosome and melanin related protein content also decrease significantly. The reason is that the Dickkopf1 protein secreted by fibroblasts in palmar plantar skin can inhibit the proliferation, development, differentiation and melanogenesis of melanocytes by inhibiting the activation of Wnt signal transduction pathway of melanocytes. In this study, the gene encoding 32-266 amino acids of Dickkop1 was obtained from human palmoplantar skin fibroblasts by reverse transcription PCR and cloned into PQE30 plasmid. The prokaryotic expression plasmid of PQE30-dkk1 was constructed and confirmed by double enzyme digestion and sequence analysis.
【学位授予单位】：第四军医大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R346

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