严重急性呼吸综合征冠状病毒S蛋白与眼部受体—血管紧张素转化酶2作用机制的研究

发布时间：2018-01-16 02:24

本文关键词：严重急性呼吸综合征冠状病毒S蛋白与眼部受体—血管紧张素转化酶2作用机制的研究　出处：《第二军医大学》2005年硕士论文　论文类型：学位论文

【摘要】：[目的] 研究SARS冠状病毒(SARS-CoV)S蛋白与眼部功能性受体—血管紧张素转化酶2(ACE2)作用的机制。[方法] 构建包含SARS-CoV S蛋白融合基因pS240-EGFP和pS666-EGFP,并在哺乳细胞中表达;半定量RT-PCR和免疫组化鉴定ACE2在结膜、角膜中的表达;酶联免疫吸附实验、冰冻切片免疫荧光法、流式细胞仪检测法、荧光显微镜检测法进行SARS-CoV S蛋白与结膜、角膜细胞的体外结合研究。[结果] 构建了融合基因pS240-EGFP和pS666-EGFP;证实ACE2在结膜、角膜有表达;结膜、角膜细胞与SARS-CoV S蛋白能够结合,但结合能力很弱。[结论] 结膜、角膜中存在ACE2,SARS-CoV S蛋白与结膜、角膜细胞结合较弱。本研究对了解SARS的病理机制及进一步研究SARS的防护和治疗具有一定的参考意义。
[Abstract]:[Objective] to study the mechanism of the action of SARS coronavirus SARS-CoV S protein on ocular functional receptor angiotensin converting enzyme 2 (ACE2). [Methods] pS240-EGFP and pS666-EGFP containing SARS-CoV S protein fusion genes were constructed and expressed in mammalian cells. The expression of ACE2 in conjunctiva and cornea was identified by semi-quantitative RT-PCR and immunohistochemistry. Enzyme linked immunosorbent assay (Elisa), frozen section immunofluorescence, flow cytometry and fluorescence microscopy were used to study the binding of SARS-CoV S protein to conjunctiva and corneal cells in vitro. [Results] the fusion genes pS240-EGFP and pS666-EGFP were constructed. It was confirmed that ACE2 was expressed in conjunctiva and cornea. Conjunctiva, corneal cells can bind to SARS-CoV S protein, but the binding ability is very weak. [Conclusion: the conjunctiva and cornea contain ACE2, SARS-CoV S protein and conjunctiva. This study is helpful to understand the pathological mechanism of SARS and to further study the protection and treatment of SARS.
【学位授予单位】：第二军医大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R373

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