發(fā)布時(shí)間：2019-03-24 10:50

【摘要】：目的： 趨化因子(chemokine)在機(jī)體的免疫反應(yīng)和疾病的發(fā)生發(fā)展過(guò)程中起著重要的作用。它們?cè)谘仔詶l件下由單核細(xì)胞、內(nèi)皮細(xì)胞及淋巴細(xì)胞表達(dá)，參與免疫活性細(xì)胞的活化和移行，從而調(diào)節(jié)免疫應(yīng)答。近年來(lái)發(fā)現(xiàn)趨化因子與中樞神經(jīng)系統(tǒng)(central nervous system，CNS)炎癥有關(guān)，如多發(fā)性硬化(mutiple sclerosis，MS)、實(shí)驗(yàn)性變態(tài)反應(yīng)性腦脊髓炎(experimental allergic encephalomyelitis，EAE)。CNS炎癥性疾病的基本病理過(guò)程是炎性細(xì)胞募集浸潤(rùn)，而趨化因子在調(diào)節(jié)淋巴細(xì)胞進(jìn)入CNS組織的過(guò)程中發(fā)揮著極其重要的作用，其中γ-干擾素誘導(dǎo)蛋白10(interferon-γ-inducible protein-10，IP-10)在CNS炎性反應(yīng)過(guò)程中的作用頗受關(guān)注。IP-10屬于趨化因子超家族CXC類(lèi)ELR-亞族，其主要作用是對(duì)多種炎癥細(xì)胞包括T淋巴細(xì)胞、單核細(xì)胞和自然殺傷細(xì)胞具有趨化活性。許多研究表明當(dāng)CNS發(fā)生感染時(shí)，小膠質(zhì)細(xì)胞開(kāi)始活化并且大量表達(dá)IP-10，IP-10介導(dǎo)炎性細(xì)胞浸潤(rùn)，從而啟動(dòng)或促進(jìn)CNS的炎性反應(yīng)進(jìn)程。為進(jìn)一步研究IP-10的生物學(xué)活性及其在CNS炎癥性疾病中的作用，本研究構(gòu)建了大鼠pET-32a(+)-IP10原核表達(dá)質(zhì)粒，并轉(zhuǎn)入大腸桿菌BL21(DE3)中表達(dá)，通過(guò)親和層析純化出IP-10融合蛋白，對(duì)其活性和趨化功能作初步的鑒定，，也為探尋以IP-10為靶治療CNS炎癥性疾病的治療新措施奠定了基礎(chǔ)。 方法： 本研究由兩部分組成：第一部分為大鼠IP-10原核表達(dá)質(zhì)粒的構(gòu)建。首先從GenBank上獲取大鼠IP-10 cDNA基因序列，使用引物設(shè)計(jì)軟件Primer Premier 5.0進(jìn)行引物設(shè)計(jì)，從中選擇最佳引物�？紤]到后續(xù)的純
[Abstract]:Objective: chemokine (chemokine) plays an important role in immune response and disease development. They are expressed by monocytes, endothelial cells and lymphocytes under inflammatory conditions, and participate in the activation and migration of immunoactive cells, thus regulating the immune response. In recent years, chemokine has been found to be associated with (central nervous system,CNS inflammation in the central nervous system, such as multiple sclerosis (mutiple sclerosis,MS) and experimental allergic encephalomyelitis (experimental allergic encephalomyelitis,). (EAE). The basic pathological process of inflammatory diseases in CNS is inflammatory cell recruitment and infiltration, and chemokine plays an important role in regulating lymphocyte entry into CNS tissue. The role of interferon-緯-induced protein 10 (interferon- 緯-inducible protein-10,IP-10) in the inflammatory process of CNS has attracted much attention. IP-10 belongs to the chemokine superfamily CXC-like ELR-subfamily. Its main function is chemotactic activity to a variety of inflammatory cells, including T lymphocytes, monocytes and natural killer cells. Many studies have shown that when CNS infection occurs, microglia begin to activate and express a large number of IP-10,IP-10-mediated inflammatory cell infiltration, thus initiating or promoting the inflammatory process of CNS. In order to further study the biological activity of IP-10 and its role in inflammatory diseases of CNS, the prokaryotic expression plasmid of rat pET-32a ()-IP10 was constructed and transformed into E. coli BL21 (DE3). The IP-10 fusion protein was purified by affinity chromatography, and its activity and chemotactic function were preliminarily identified. It also laid a foundation for exploring new therapeutic measures for treating inflammatory diseases of CNS with IP-10 as target. Methods: this study consists of two parts: the first part is the construction of prokaryotic expression plasmid of rat IP-10. At first, the sequence of rat IP-10 cDNA gene was obtained from GenBank. Primer design software Primer Premier 5.0 was used to design the primer, and the best primer was selected from the primer design software Primer Premier 5.0. Considering the subsequent purity,
【學(xué)位授予單位】：四川大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類(lèi)號(hào)】：R346

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