本文關(guān)鍵詞：人海綿狀血管瘤內(nèi)皮細胞特性研究和人海綿狀血管瘤動物模型的建立　出處：《中國協(xié)和醫(yī)科大學(xué)》2005年博士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 血管瘤 內(nèi)皮細胞 血管生成 內(nèi)質(zhì)網(wǎng) 表型 功能特性 基因

【摘要】：血管瘤是最常見的嬰幼兒良性血管增生或血管畸形，好發(fā)于顏面部，常因引起嚴(yán)重的面部畸形而影響患者身心健康。盡管血管瘤的形成可能與活躍的血管生成有關(guān)，但是其發(fā)病機制還不明了。而且目前對血管瘤的治療尚缺乏安全有效的藥物，因此，研究血管瘤內(nèi)皮細胞的表型和功能特性，，并建立一個血管瘤實驗動物模型對血管瘤發(fā)病機制的研究和臨床治療藥物的篩選有非常重要的意義。 目的 研究人海綿狀血管瘤內(nèi)皮細胞在形態(tài)、表型、功能和基因方面的改變，探索海綿狀血管瘤的發(fā)生機制；建立人海綿狀血管瘤的實驗動物模型，并篩選對海綿狀血管瘤有效的治療藥物。 方法 從人海綿狀血管瘤組織分離、純化血管瘤內(nèi)皮細胞。應(yīng)用免疫熒光染色和流式細胞術(shù)鑒定其內(nèi)皮細胞標(biāo)志性分子的表達。應(yīng)用透射電子顯微鏡觀察海綿狀血管瘤內(nèi)皮細胞的超微結(jié)構(gòu)的變化；應(yīng)用流式細胞術(shù)和免疫細胞化學(xué)方法分析海綿狀血管瘤內(nèi)皮細胞的表型特征；應(yīng)用Matrigel比較血管瘤內(nèi)皮細胞與正常血管內(nèi)皮細胞形成毛細血管樣網(wǎng)絡(luò)能力的差別：應(yīng)用RT-PCR研究海綿狀血管瘤內(nèi)皮細胞上VEGF及其受體、Ang-1、Ang-2和Tie2的表達。海綿狀血管瘤內(nèi)皮細胞表達和分泌的基質(zhì)金屬蛋白酶分別通過明膠酶譜法、RT-PCR、Western blot和細胞免疫熒光染色進行分析。應(yīng)用酶聯(lián)免疫吸附測定檢查海綿狀血管瘤內(nèi)皮細胞釋放的U-PA、PAI-1和TF。通過將熒光染料標(biāo)記的血小板與內(nèi)皮細胞共培養(yǎng)比較血管瘤內(nèi)皮細胞和正常內(nèi)皮細胞與血小板粘附能力的差別。此外，應(yīng)用RT-PCR方法，比較252-基因在血管瘤內(nèi)皮細胞，正常血管內(nèi)皮細胞和腫瘤血管內(nèi)皮細胞的表達差別，并應(yīng)用透射電子顯微鏡觀察252-基因轉(zhuǎn)染后對人海綿狀血管瘤內(nèi)皮細胞超微結(jié)構(gòu)異常的影響。
[Abstract]:Hemangioma is the most common benign vascular hyperplasia or vascular malformation in infants. Physical and mental health is often affected by severe facial malformations, although the formation of hemangiomas may be associated with active angiogenesis. However, the pathogenesis of hemangioma is not clear, and the current treatment of hemangioma is still lack of safe and effective drugs, therefore, to study the phenotypic and functional characteristics of endothelial cells of hemangioma. The establishment of an experimental animal model of hemangioma is of great significance for the study of the pathogenesis of hemangioma and the screening of clinical therapeutic drugs. Objective to study the morphological phenotypic functional and genetic changes of human cavernous hemangioma endothelial cells and to explore the pathogenesis of cavernous hemangioma. To establish an experimental animal model of human cavernous hemangioma and to screen effective drugs for treatment of cavernous hemangioma. Methods Human cavernous hemangioma was isolated. The expression of iconic molecules in endothelial cells was identified by immunofluorescence staining and flow cytometry. The ultrastructural changes of endothelial cells in cavernous hemangioma were observed by transmission electron microscope (TEM). The phenotypic characteristics of endothelial cells in cavernous hemangioma were analyzed by flow cytometry and immunocytochemistry. Matrigel was used to compare the ability of angioma endothelial cells and normal vascular endothelial cells to form capillary networks. RT-PCR was used to study VEGF and its receptor on endothelial cells of cavernous hemangioma. Expression of Ang-1 Ang-2 and Tie2. The matrix metalloproteinases secreted by endothelial cells of cavernous hemangioma were detected by reverse transcriptase polymerase chain reaction (RT-PCR). Western blot and cellular immunofluorescence staining were used to detect U-PA released from endothelial cells of cavernous hemangioma by enzyme-linked immunosorbent assay (Elisa). PAI-1 and TF. The difference of adhesion between endothelial cells of hemangioma and that of normal endothelial cells was compared by co-culture of fluorescent dye labeled platelets and endothelial cells. To compare the expression of 252-gene in angioma endothelial cells, normal vascular endothelial cells and tumor vascular endothelial cells by RT-PCR. The effect of 252-gene transfection on the ultrastructure of human cavernous hemangioma endothelial cells was observed by transmission electron microscope.
【學(xué)位授予單位】：中國協(xié)和醫(yī)科大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2005
【分類號】：R-332

【引證文獻】

相關(guān)期刊論文 前1條

1 楊壯群;馬建明;屠軍波;王正輝;宋勇;虎小毅;徐泉;李鵬;;脂質(zhì)體魚肝油酸鈉對人血管瘤內(nèi)皮細胞的作用[J];中國口腔頜面外科雜志;2007年06期

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