本文選題：細(xì)胞因子信號(hào)抑制蛋白3 + RNA干擾　； 參考：《吉林大學(xué)》2012年碩士論文

【摘要】：支氣管哮喘是以Th2細(xì)胞增多占優(yōu)勢(shì)，同時(shí)伴隨肥大細(xì)胞、嗜酸性粒細(xì)胞聯(lián)合浸潤(rùn)為特征的一種慢性疾病，Th1/Th2的比例失衡和以Th2型細(xì)胞為主的免疫應(yīng)答在支氣管哮喘的發(fā)病機(jī)制中起到重要作用。SOCS-3是JAK/STAT信號(hào)通路的反饋抑制因子，又被稱為細(xì)胞的“分子剎車”。SOCS-3與多種疾病的發(fā)生發(fā)展有著密切關(guān)系，同時(shí)亦是多條信號(hào)通路的交界點(diǎn)，從而其調(diào)控、作用機(jī)制復(fù)雜。SOCS-3不僅是細(xì)胞因子的調(diào)節(jié)者，而且在調(diào)控細(xì)胞分化和決定細(xì)胞分化方面起著重要作用。SOCS-3在Th2細(xì)胞中高表達(dá)，并可被許多炎癥或抗炎癥因子誘導(dǎo)生成，，同時(shí)能反作用于Th細(xì)胞，影響其分化。 本實(shí)驗(yàn)根據(jù)小鼠SOCS3的mRNA序列設(shè)計(jì)3條shRNA序列，化學(xué)合成后與PLL3.7質(zhì)粒連接，產(chǎn)生PLL3.7-SOCS-3-shRNA慢病毒重組質(zhì)粒，經(jīng)體外轉(zhuǎn)染小鼠B16黑色素瘤細(xì)胞鑒定其沉默效率，選取沉默效率最高者與慢病毒包裝質(zhì)粒共轉(zhuǎn)染293T細(xì)胞，經(jīng)收集細(xì)胞上清和低溫超高速離心，最終獲得高濃度的慢病毒載體。本實(shí)驗(yàn)構(gòu)建SOCS-3RNA干擾慢病毒載體，以期為后續(xù)的支氣管哮喘的研究與治療提供實(shí)驗(yàn)工具。
[Abstract]:Bronchial asthma is dominated by the proliferation of Th2 cells, accompanied by mast cells.The imbalance of Th1 / Th2 ratio in a chronic disease characterized by eosinophilic granulocyte infiltration and the immune response with Th2 type cells play an important role in the pathogenesis of bronchial asthma. SOCS-3 is a feedback inhibitor of JAK/STAT signaling pathway.SOCS-3, also known as cell "molecular brake", is closely related to the occurrence and development of many diseases, and it is also the junction point of many signal pathways. Therefore, the complex mechanism of SOCS-3 is not only the modulator of cytokines, but also the junction of many signal pathways.SOCS-3 plays an important role in regulating cell differentiation and determining cell differentiation. SOCS-3 is highly expressed in Th2 cells and can be induced by many inflammatory or anti-inflammatory factors.In this study, three shRNA sequences were designed according to the mRNA sequence of mouse SOCS3, which were chemically synthesized and ligated with PLL3.7 plasmid to produce PLL3.7-SOCS-3-shRNA lentivirus recombinant plasmid. The recombinant plasmid was transfected into murine B16 melanoma cells in vitro to identify its silencing efficiency.The highest silencing efficiency and lentivirus packaging plasmid were cotransfected into 293T cells. After supernatant and cryogenic centrifugation, a high concentration of lentivirus vector was obtained.In this study, SOCS-3RNA interference lentivirus vector was constructed to provide experimental tools for the further study and treatment of bronchial asthma.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R562.25

【參考文獻(xiàn)】

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1 王藝穎 ,高紅亮 ,張毅;細(xì)胞因子發(fā)信號(hào)阻抑蛋白SOCS3作用機(jī)制及其在相關(guān)疾病中的研究[J];生命的化學(xué);2005年02期

2 祖瑩;李成榮;李德發(fā);鄧?yán)^巋;鄭躍杰;;細(xì)胞因子信號(hào)抑制因子在支氣管哮喘T_H1／T_H2細(xì)胞失衡中的作用[J];中華微生物學(xué)和免疫學(xué)雜志;2006年06期

本文編號(hào)：1761100