人皮肤上皮化中PAI-3表达及作用的初步研究

发布时间：2018-01-18 00:25

本文关键词：人皮肤上皮化中PAI-3表达及作用的初步研究　出处：《第三军医大学》2005年硕士论文　论文类型：学位论文

【摘要】：皮肤位于机体外环境和内环境的界面上,是机体重要的屏障保护器官,其表皮绝大部分由角质形成细胞(KC)组成。在皮肤上皮化中,蛋白酶是一类重要的调节分子。PAI-3 又称为蛋白C 抑制剂(protin C inhibitor PCI),是纤溶酶原激活剂及其抑制剂(Plasminogen Activator/ Plasminogen Activator inhibitor,PA/PAI)系统的重要成员。新近的研究表明:PAI-3 在人的肝脏、胰腺、泌尿生殖系统、血小板等多个器官和组织中表达,在凝血、纤溶、细胞迁移、生殖及肿瘤侵袭转移中具有重要作用。然而PAI-3 是否参与人的皮肤上皮化,目前国内外尚未见报道。本实验旨在研究正常成人皮肤组织及培养分化KC 中和胚胎生长发育过程表皮KC 中的PAI-3 的表达及作用,希望有助于阐明PAI-3 在皮肤上皮化中的功能,为进一步研究PAI-3 在皮肤上皮化中的机制及各种皮肤疾病提供实验基础。方法: 采用免疫组织化学(Immunohistochemistry,IHC)、(Immunocytochemistry,ICC)和逆转录-聚合酶链反应(reverse transcription PCR,RT-PCR ) 技术及KC角质壳的提取,从三个方面研究了PAI-3 在人皮肤上皮化中的表达及作用。首先对PAI-3 在正常成人皮肤组织中的表达及作用进行初步研究后,在高钙条件下培养的分化KC 中的表达及作用进行了探讨,最后对其在胚胎生长发育过程皮肤表皮KC 中的表达进行了检测。结果: 1、PAI-3 在正常成人皮肤组织中表达,并且主要位于分化程度较高的KCs中。 2、PAI-3 存在终末分化KCs 角质壳中。 3、PAI-3 蛋白及基因的转录在分化KC 中表达,并随着分化的增加,表达增强,但在后期,其蛋白表达及基因的转录开始降低。 4、PAI-3 蛋白在增殖KC 中主要聚集在细胞核膜周围,而在分化KC 中主要位于细胞的胞浆中。 5、PAI-3 蛋白在人胚胎主要位于分化较高的表皮KC 中,并且随着胚胎的生长发育,其表达增强,表达的高峰在中期,晚期表达开始降低。 6、PAI-3 基因的转录随着人胚胎的生长发育,其表达增强,表达的高峰在中期,但在晚期仍然维持在较高的水平。结论: 1、PAI-3 在人皮肤表皮组织中表达,且主要位于分化程度较高的表皮KC 中,其存在终末分化KCs 角质壳中。 2、随着KC 的分化,PAI-3 蛋白及基因转录增强,并且由核膜向胞浆转移。
[Abstract]:The skin is located on the interface of the external environment and the internal environment of the body. It is an important protective organ of the body. Most of its epidermis is composed of keratinocytes (KCs) and is in the process of skin epithelialization. Protease is an important regulatory molecule. PAI-3 is also known as protein C inhibitor protin C inhibitor. Plasminogen activator / Plasminogen Activator inhibitor. A key member of the PAR / PAI system. Recent studies have shown that WPAI-3 is expressed in many organs and tissues, such as human liver, pancreas, urogenital system, platelets, and so on, and is expressed in coagulation and fibrinolysis. Cell migration, reproduction and tumor invasion and metastasis play important roles. However, whether PAI-3 is involved in skin epithelization has not been reported at home and abroad. The purpose of this study was to investigate the expression and role of PAI-3 in normal adult skin tissues and cultured and differentiated KC and the epidermis KC during embryonic growth and development. It is hoped that it will be helpful to clarify the function of PAI-3 in skin epithelization and provide experimental basis for further study on the mechanism of PAI-3 in skin epithelization and various skin diseases. Methods: Immunocytochemistry of IHC was used in this study. ICC), reverse transcription PCR RT-PCR and KC keratinocyte extraction. The expression and role of PAI-3 in human skin epithelization were studied from three aspects. Firstly, the expression and role of PAI-3 in normal adult skin were studied. The expression and function of differentiated KC in high calcium culture were studied. Finally, the expression of KC in skin epidermis during embryonic growth and development was detected. 1PAI-3 was expressed in normal adult skin tissues, and was mainly located in highly differentiated KCs. 2 the terminal differentiation of KCs keratinocytes was found in PAI-3. 3The transcription of PAI-3 protein and gene was expressed in differentiated KC and increased with the increase of differentiation, but the protein expression and gene transcription began to decrease at the later stage. (4) PAI-3 protein mainly accumulates around the nuclear membrane in proliferative KC and mainly in cytoplasm in differentiated KC. 5PAI-3 protein was mainly located in the highly differentiated epidermis KC of human embryos, and the expression of PAI-3 protein increased with the development of embryos, the peak of expression began to decrease in the middle stage and the late stage. The expression of PAI-3 gene increased with the development of human embryo, and the expression peak was in the middle stage, but remained at a high level at the late stage. Conclusion: (1) PAI-3 was expressed in human skin epidermis and was mainly located in the highly differentiated epidermis KC, which had terminal differentiation in KCs keratinocytes. 2, with the differentiation of KC, PAI-3 protein and gene transcription increased, and transferred from nuclear membrane to cytoplasm.
【学位授予单位】：第三军医大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R329

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