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  本文關(guān)鍵詞：人皮膚上皮化中PAI-3表達(dá)及作用的初步研究　出處：《第三軍醫(yī)大學(xué)》2005年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： PAI-3 PCI 上皮化 角質(zhì)殼 Ca2+ KC 分化 人胚胎 PA/PAI 系統(tǒng)

【摘要】：皮膚位于機(jī)體外環(huán)境和內(nèi)環(huán)境的界面上,是機(jī)體重要的屏障保護(hù)器官,其表皮絕大部分由角質(zhì)形成細(xì)胞(KC)組成。在皮膚上皮化中,蛋白酶是一類重要的調(diào)節(jié)分子。PAI-3 又稱為蛋白C 抑制劑(protin C inhibitor PCI),是纖溶酶原激活劑及其抑制劑(Plasminogen Activator/ Plasminogen Activator inhibitor,PA/PAI)系統(tǒng)的重要成員。新近的研究表明:PAI-3 在人的肝臟、胰腺、泌尿生殖系統(tǒng)、血小板等多個器官和組織中表達(dá),在凝血、纖溶、細(xì)胞遷移、生殖及腫瘤侵襲轉(zhuǎn)移中具有重要作用。然而PAI-3 是否參與人的皮膚上皮化,目前國內(nèi)外尚未見報道。 本實(shí)驗(yàn)旨在研究正常成人皮膚組織及培養(yǎng)分化KC 中和胚胎生長發(fā)育過程表皮KC 中的PAI-3 的表達(dá)及作用,希望有助于闡明PAI-3 在皮膚上皮化中的功能,為進(jìn)一步研究PAI-3 在皮膚上皮化中的機(jī)制及各種皮膚疾病提供實(shí)驗(yàn)基礎(chǔ)。 方法: 采用免疫組織化學(xué)(Immunohistochemistry,IHC)、(Immunocytochemistry,ICC)和逆轉(zhuǎn)錄-聚合酶鏈反應(yīng)(reverse transcription PCR,RT-PCR ) 技術(shù)及KC角質(zhì)殼的提取,從三個方面研究了PAI-3 在人皮膚上皮化中的表達(dá)及作用。首先對PAI-3 在正常成人皮膚組織中的表達(dá)及作用進(jìn)行初步研究后,在高鈣條件下培養(yǎng)的分化KC 中的表達(dá)及作用進(jìn)行了探討,最后對其在胚胎生長發(fā)育過程皮膚表皮KC 中的表達(dá)進(jìn)行了檢測。結(jié)果: 1、PAI-3 在正常成人皮膚組織中表達(dá),并且主要位于分化程度較高的KCs中。 2、PAI-3 存在終末分化KCs 角質(zhì)殼中。 3、PAI-3 蛋白及基因的轉(zhuǎn)錄在分化KC 中表達(dá),并隨著分化的增加,表達(dá)增強(qiáng),但在后期,其蛋白表達(dá)及基因的轉(zhuǎn)錄開始降低。 4、PAI-3 蛋白在增殖KC 中主要聚集在細(xì)胞核膜周圍,而在分化KC 中主要位于細(xì)胞的胞漿中。 5、PAI-3 蛋白在人胚胎主要位于分化較高的表皮KC 中,并且隨著胚胎的生長發(fā)育,其表達(dá)增強(qiáng),表達(dá)的高峰在中期,晚期表達(dá)開始降低。 6、PAI-3 基因的轉(zhuǎn)錄隨著人胚胎的生長發(fā)育,其表達(dá)增強(qiáng),表達(dá)的高峰在中期,但在晚期仍然維持在較高的水平。 結(jié)論: 1、PAI-3 在人皮膚表皮組織中表達(dá),且主要位于分化程度較高的表皮KC 中,其存在終末分化KCs 角質(zhì)殼中。 2、隨著KC 的分化,PAI-3 蛋白及基因轉(zhuǎn)錄增強(qiáng),并且由核膜向胞漿轉(zhuǎn)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.
【學(xué)位授予單位】：第三軍醫(yī)大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2005
【分類號】：R329

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本文編號：1438663