SOCS-3基因沉默对哮喘小鼠气道炎症的影响

发布时间：2018-06-02 00:17

本文选题：SOCS-3 + 支气管哮喘　；参考：《吉林大学》2013年硕士论文

【摘要】：支气管哮喘的特征性表现有慢性气道炎症、可逆性气道阻塞、气道高反应性（AHR，非特异性刺激后因气流阻塞增强而产生）、黏液过度分泌及上皮的纤维化。该类患者的支气管周围可见大量嗜酸性粒细胞、淋巴细胞的浸润。Th2细胞是浸润支气管哮喘病人气道的主要淋巴细胞亚群，其分泌的细胞因子（IL-4、IL-5、IL-13）与气道嗜酸性粒细胞增多、气道高反应性及血清IgE增加等一系列症状的发生密切相关。这些细胞因子首先与细胞表面的受体结合，然后激活信号转导复合物，包括JAK和STAT，以及Ras-细胞外信号调解激酶，继而产生效应。 SOCS是一类抑制JAK-STAT信号通路的蛋白家族分子，该蛋白能够调解Th细胞的分化。其中Th2细胞高表达SOCS-3蛋白，在Th2细胞中，IL-12介导的STAT活化途径被抑制。SOCS-3抑制该途径是通过结合IL-12受体的β亚基中的STAT-4的位点来实现的。在支气管哮喘或者非特异性皮炎患者体内，，T细胞的SOCS-3转录物的表达水平与血清IgE水平及疾病的严重程度密切相关。T细胞中的SOCS-3基因的表达导致OVA诱导的过敏性哮喘小鼠模型的Th2型细胞分化、气道的嗜酸性粒细胞增加以及AHR增强。有实验证明T细胞中缺乏SOCS-3基因的小鼠的Th1和Th2细胞分化减少，向Th3细胞分化增加，从而导致转化生长因子β（TGF-β）产生增加。且已有研究表明，SOCS-3能够抑制CD4+CD25+Foxp3+调节性T细胞的增殖及其功能。因此，下调T细胞中的SOCS-3的表达对支气管哮喘的的治疗可能有作用。本研究是将课题组前期已设计并鉴定其沉默效率的特异性SOCS-3siRNA慢病毒载体，经体内转染BALB/c哮喘小鼠模型，留取肺组织标本及血清，应用HE染色、刚果红染色、ELISA、免疫组织化学、RT-PCR和Western Blot等实验方法比较SOCS-3siRNA慢病毒载体体内转染对哮喘小鼠气道炎症及Th1/Th2比例平衡的影响。本实验采用基因沉默的方法，沉默SOCS-3基因，达到改善哮喘小鼠模型的哮喘症状的目的，以期为支气管哮喘的临床研究与治疗提供理论及实验依据。
[Abstract]:The characteristic manifestations of bronchial asthma include chronic airway inflammation reversible airway obstruction airway hyperresponsiveness AHRs increased airflow obstruction after nonspecific stimulation mucus hypersecretion and epithelial fibrosis. A large number of eosinophilic granulocytes were found around the bronchi of these patients. The infiltration of lymphocytes. Th2 cells were the main lymphocyte subsets in the airway of asthmatic patients. The cytokines secreted IL-4 / IL-5 / IL-13) and the number of eosinophils in the airway were increased. A series of symptoms such as airway hyperresponsiveness and increased serum IgE are closely related. These cytokines first bind to receptors on the cell surface and then activate signal transduction complexes, including JAK and stat, as well as Ras-extracellular signal-mediated kinases. SOCS is a family of proteins that inhibit the JAK-STAT signaling pathway and mediate the differentiation of Th cells. The overexpression of SOCS-3 protein in Th2 cells was inhibited by the inhibition of STAT activation pathway mediated by IL-12 in Th2 cells. SOCS-3 inhibited this pathway by binding the STAT-4 site in 尾 subunit of IL-12 receptor. Expression of SOCS-3 transcripts in T cells of patients with bronchial asthma or nonspecific dermatitis is closely related to serum IgE level and severity of disease. The expression of SOCS-3 gene in T cells leads to OVA induced allergic asthma. Differentiation of Th2 type cells in asthmatic mice, Airway eosinophilic granulocytes increased and AHR increased. Some experiments have proved that the differentiation of Th1 and Th2 cells in mice lacking SOCS-3 gene in T cells decreased, and the differentiation to Th3 cells increased, which resulted in the increase of TGF- 尾 production. It has been shown that SOCS-3 can inhibit the proliferation and function of CD4 CD25 Foxp3 regulatory T cells. Therefore, down-regulation of SOCS-3 expression in T cells may play a role in the treatment of bronchial asthma. In this study, we designed and identified the specific SOCS-3siRNA lentivirus vector with silencing efficiency, transfected BALB/c asthma mouse model in vivo, collected lung tissue and serum, and used HE staining. The effects of SOCS-3siRNA lentivirus vector transfection on airway inflammation and Th1/Th2 balance in asthmatic mice were compared by Congo red staining Elisa immunohistochemistry RT-PCR and Western Blot. In this study, gene silencing was used to silence the SOCS-3 gene to improve the asthmatic symptoms in mice model of asthma, in order to provide theoretical and experimental basis for the clinical study and treatment of bronchial asthma.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R562.25

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