SOCS3蛋白与甲状腺相关性眼病的相关性研究

发布时间：2018-05-11 06:03

本文选题：甲状腺相关性眼病 + 细胞因子信号转导抑制分子-3　；参考：《华中科技大学》2015年博士论文

【摘要】：研究背景和目的：甲状腺相关性眼病是与Graves病密切相关的眼眶组织炎症,发病率居成年人眼眶疾病之首,大量细胞因子参与疾病过程,引起组织重构,但确切的发病机制尚不清楚。细胞因子信号转导抑制分子-3(suppressor of cytokine signaling-3, SOCS3)作为细胞因子重要的负调控因子,在多种炎症性疾病发生、发展过程中起到重要的作用。本研究探讨SOCS3蛋白与甲状腺相关性眼病的关系。方法： 1.取甲状腺相关性眼病患者和正常人的眶脂肪组织,检测眶脂肪组织中是否表达SOCS3蛋白。使用免疫荧光法。 2.分别由甲状腺相关性眼病患者和正常人眼眶结缔组织分离培养成纤维细胞,检测两组眼眶成纤维细胞中SOCS3mRNA和蛋白质的表达差异。使用real-time PCR和Western blot法。 3.收集270例Graves病患者病例资料及外周静脉血。将所有患者根据有无眼病分为Graves病伴有眼病组(114例)和Graves病不伴眼病组(156例)。分别取两组患者外周血单核细胞(PBMC),建立EB病毒转化的类淋巴母细胞系(Epstein-Barr virus lymphoblastoid cell lines, EBV-LCLs)。检测两组患者EBV-LCLs中SOCS3mRNA和蛋白质的表达差异。使用real-time PCR和Western blot方法。对所有患者SOCS3基因5个位点(rs12952093, rs4969170, rs4969168, rs4969169and rs2280148)进行测序分型,采用连接酶检测反应-PCR (LDR-PCR)基因测序分型技术。多因素Logistic回归分析SOCS3基因各位点基因型与甲状腺相关性眼病间的关系。为进一步验证与眼病有关的基因型是否影响基因功能,研究继续观察该基因型是否影响SOCS3表达。使用real-time PCR和Western blot法。结果： 1.甲状腺相关性眼病患者眶脂肪组织表达SOCS3蛋白非常明显,正常人组织中微量表达。 2.甲状腺相关性眼病患者眶成纤维细胞中SOCS3mRNA和蛋白质的水平较正常人显著增高(p0.05)。 3. Graves病伴眼病患者EBV-LCLs中SOCS3mRNA和蛋白的表达水平较Graves病无眼病患者显著增高(p0.05)。 4.校正年龄、性别、BMI,吸烟,甲状腺肿,甲状腺结节,黏液性水肿,Graves病治疗(药物,放射性治疗,甲状腺切除术)以及高血压这些因素后,经多因素Logistic回归分析,SOCS3rs4969170AA基因型与甲状腺相关性眼病风险增高有关(OR-3.5,95%CI1.6to7.5, p=0.001).而SOCS3rs12952093, rs4969168,rs4969169及rs2280148与甲状腺相关性眼病的关联均没有达到统计学显著性水平。SOCS3rs4969170AA基因型甲状腺相关性眼病患者EBV-LCLs中SOCS3mRNA和蛋白表达水平较GG型眼病患者显著增高。结论： SOCS3蛋白与甲状腺相关性眼病有关。甲状腺相关性眼病患者中SOCS3rs4969170可能是功能性单核苷酸多态性(Single nucleotide polymorphism, SNP),该位点AA基因型可能通过增加SOCS3表达,与甲状腺相关性眼病风险增加有关。
[Abstract]:Background and objectives of the study: Thyroid associated ophthalmopathy is a kind of orbital inflammation closely related to Graves's disease. The incidence rate is the highest among adult orbital diseases. A large number of cytokines participate in the process of the disease and cause tissue remodeling, but the exact pathogenesis is not clear. Cytokine signal transduction suppressor of cytokine signaling-3 (SOCS3), as an important negative regulator of cytokines, plays an important role in the occurrence and development of various inflammatory diseases. This study was to investigate the relationship between SOCS3 protein and thyroid associated ophthalmopathy. Methods: 1. The orbital adipose tissues of patients with thyroid associated ophthalmopathy and normal subjects were taken to detect the expression of SOCS3 protein in orbital adipose tissue. Immunofluorescence method was used. 2. The fibroblasts were isolated from the orbital connective tissue of the patients with thyroid associated ophthalmopathy and the normal subjects. The expression of SOCS3mRNA and protein in the orbital fibroblasts of the two groups were detected. Real-time PCR and Western blot methods were used. 3. Data of 270 patients with Graves's disease and peripheral venous blood were collected. All the patients were divided into two groups according to the presence or absence of ophthalmopathy: Graves's disease with ophthalmopathy (n = 114) and Graves's disease without ophthalmopathy (n = 156). Epstein-Barr virus lymphoblastoid cell lines, EBV-LCLs were isolated from peripheral blood monocytes of two groups of patients and transformed into Epstein-Barr virus lymphoblastoid cell lines, EBV-LCLs1 cell lines. The expression of SOCS3mRNA and protein in EBV-LCLs of the two groups were detected. Use real-time PCR and Western blot methods. Five loci of SOCS3 gene (rs12952093, rs4969170, rs4969168, rs4969169and rs2280148) were sequenced. Multivariate Logistic regression analysis showed the relationship between SOCS3 gene genotypes and thyroid associated ophthalmopathy. To further verify whether the gene function was affected by the genotype associated with eye disease, the study continued to observe whether the genotype affected the expression of SOCS3. Real-time PCR and Western blot methods were used. Results: 1. The expression of SOCS3 protein in orbital adipose tissue of thyroid associated ophthalmopathy patients was very obvious, and the expression of SOCS3 protein in normal tissue was very small. 2. The levels of SOCS3mRNA and protein in orbital fibroblasts in patients with thyroid associated ophthalmopathy were significantly higher than those in normal controls (P 0.05). 3. The expression of SOCS3mRNA and protein in EBV-LCLs in patients with Graves disease and ophthalmopathy was significantly higher than that in patients without Graves disease. 4. Adjusted for age, sex, BMI, smoking, goiter, thyroid nodule, mucinous edema, Graves' disease treatment (drugs, radiation therapy, thyroidectomy) and hypertension. Multivariate Logistic regression analysis showed that SOCS3rs4969170AA genotype was associated with increased risk of thyroid associated ophthalmopathy. The correlation between SOCS3rs12952093, rs4969168rs4969169 and rs2280148 was not statistically significant. The expression of SOCS3mRNA and protein in EBV-LCLs of patients with thyroid associated ophthalmopathy genotype SOCS3rs4969170AA was significantly higher than that of patients with GG type ophthalmopathy. Conclusion: SOCS3 protein is associated with thyroid associated ophthalmopathy. SOCS3rs4969170 may be a functional single nucleotide polymorphism (SNPs) in patients with thyroid associated ophthalmopathy. The AA genotype at this locus may be associated with an increased risk of thyroid associated ophthalmopathy by increasing the expression of SOCS3.
【学位授予单位】：华中科技大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R777.5

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