免疫抑制剂冲击治疗影响甲状腺相关性眼病患者泪液蛋白质组学的研究

It is common to find ocular surface damages in TAO patients, but its pathogenesis is unknown. In this project, we plan to use comparative proteomics of the normal group and TAO patients before / after immunosuppressive agents pulse therapy to analyze tear protein expression profile;In the project,we choose capillary high performance liquid chromatography(HPLC) as separation system, choose Q-Exactive mass spectrometer as a means of detection,choose the label free quantitative proteomic, Perseus statistics and bioinformatics to analyze.Then we will check differentially expressed proteins by western blot tests. The research will be expected: 1 to get the differences in proteome expression protein expression quantity, type and amount of information in tears between TAO and normal people; 2 to obtain changes of protein expression differences of TAO tear before and after immunosuppressive agents pulse therapy.The meaning of the project: 1 to make a new explanation of the molecular pathogenesis of ocular surface damage of TAO from tear proteomics level; 2 to explore immune inhibitors of protein expression differences of effects in TAO tears, expected to find drug targets on ocular surface; 3 to have the potential to provide important inspiration in finding molecular markers of diagnosis and therapy evaluation of TAO.

常见眼眶病甲状腺相关性眼病（Thyroid associated ophthalmopathy，TAO）眼表损害多见，其确切发生机制不明。本项目运用比较蛋白质组学对正常人组、TAO患者免疫抑制剂冲击治疗前/后的泪液蛋白质表达谱进行分析：通过毛细管高效液相色谱系统分离，用Q-Exactive质谱仪检测，进行非标记定量蛋白质组学研究，最后行Perseus统计学和生物信息学分析，以免疫印迹法进行差异表达蛋白质验证。预期研究将：1. 获得TAO和正常人泪液蛋白质组差异表达蛋白的数量、种类及表达量信息；2. 获得免疫抑制剂冲击治疗前后，TAO泪液差异表达蛋白的变化情况。本项目旨在：1. 从泪液蛋白质组学层面，对TAO眼表损害的分子病理机制作出新解释；2. 探究免疫抑制剂对TAO泪液差异表达蛋白的影响，期望能发现药物在眼表可能的作用靶点；3. 为发现TAO诊断及治疗评估的分子标志物提供重要启示。

常见眼眶病甲状腺相关性眼病（Thyroid associated ophthalmopathy，TAO）眼表损害多见，其确切发生机制不明。本项目运用比较蛋白质组学对正常人组、TAO患者免疫抑制剂冲击治疗前/后的泪液蛋白质表达谱进行分析：通过毛细管高效液相色谱系统分离，用Q-Exactive质谱仪检测，进行非标记定量蛋白质组学研究，最后行Perseus统计学和生物信息学分析，以免疫印迹法进行差异表达蛋白质验证。本项目重要结果为：①首次运用基于 LC-MS／MS 的非标记定量蛋白质组学技术分析正常人和 TAO.患者冲击前后泪液的相关差异蛋白，鉴定出正常人和 TAO 患者泪液中差异超过 4 倍的上调蛋白 99 个、下调蛋白 4 个，TAO 患者冲击治疗前后泪液中差异超过 2 倍的上调蛋白 11 个，下调蛋白 9 个。这一结果提示非标记定量蛋白质组学是研究 TAO 泪液差异蛋白质的有效手段。②应用生物信息学方法分析和注释这些差异蛋白，发现他们参与代谢等生物过程，涉及到蛋白结合等分子功能，并富集于细胞等细胞成分；这些蛋白还参与多个信号转导通路，联系密切的有补体与凝血级联通路等，提示 TAO 患者眼表损害的机制与该通路关系密切，即和 TAO 患者眼表炎症及免疫反应关系密切。这些结果可为今后 TAO 眼表损害发病机制及发病标志物的研究奠定基础。