从T淋巴细胞及PPARγ对眶成纤维细胞重构的调控途径研究消瘿合剂对甲状腺相关眼病的作用机制

Thyroid associated ophthalmopathy (TAO) characterized by expanded volume of the orbital tissues is an autoimmune inflammatory disease. Thyroid stimulating hormone receptor (TSHR) mediated the autoimmune ophthalmopathy diseases. Serious threat to the living quality of TAO patients. Orbital fibroblasts(OF) are thought to be a key target and effector cell in the pathogenesis of TAO. The activation of T lymphocyte lead to the beginning, then OF proliferation, differentiation into adipocytes. OF reconstruction is the main pathological mechanisms. The secretory function of fat cell accelerate the pathological progress. Our previous study demonstrated: Xiaoying mixture can effectively reduce T lymphocyte immune inflammatory response, and relieve OF reconstruction. The contents of this research are as follows : ① The effects of Xiaoying mixture on the expressions of MP and insulin-like growth factor 1 (IGF-1) by T lymphocytes and the activation of OF are examined, and the correlation between MP and IGF-1 expressions and the activation of OF is analyzed; ② the mechanism of OF activation regulated by MP expressed by T cell ; the mechanism of OF activation regulated by based on the T cell pathway. ③ The effects of Xiaoying mixture on the expressions of IGF-1 and peroxisome proliferator-activated receptor-gama (PPAR γ) derived from in the orbital fatty connective tissues. This research will confirm that the mechanism of Xiaoying mixture on anti- orbital fibrosis reconstruction. Whether the Xiaoying mixture can inhibite the MP、IGF-1 expression and reduce PPARγ expression of the regulational roles on fat cell actviation, and inhibiting the adipocyte differentiation. This study will in further elucidate the modern biological mechanimsm of Chinese herbs on TAO.

甲状腺相关眼病(TAO)是严重威胁患者生存质量的自身免疫性致盲性眼眶疾病。T淋巴细胞活化分泌微粒(MP)为TAO的启动因素，继之的眼眶成纤维细胞增殖、分化重构为脂肪细胞是TAO主要病理机制。胰岛素样生长因-1(IGF-1)、过氧化物酶体增殖物激活受体γ(PPARγ)是调节脂肪细胞分化的重要信号分子，与TAO密切相关。前期研究发现“消瘿合剂”能抑制TAO免疫炎性反应，改善眶成纤维细胞(OF)分化。本研究拟开展：①建立TAO小鼠模型，观察消瘿合剂对T细胞分泌MP、IGF-1及OF活化的影响，并观察MP与OF活化的相关性；②观察T淋巴细胞株表达的MP对OF活化的影响及消瘿合剂的干预作用；③体外培养人OF，观察消瘿合剂如何通过IGF-1、PPARγ干预脂肪细胞分化。以证实“消瘿合剂可以通过调节MP来抑制T淋巴细胞、OF活化，同时通过PPARγ途径干预脂肪细胞分化，来调控OF重构的上下游环节”的假说

研究基于甲状腺相关眼病(TAO)发展过程中PPARγ、IGF-1对眶成纤维细胞的作用，与“消瘿合剂”对甲状腺相关眼病的临床疗效。通过体外、动物实验分析消瘿合剂对甲状腺相关眼病眶成纤维细胞分化进程调节的机制。项目开展顺利。主要内容包括：①消瘿合剂含药血清对手术取样的体外培养TAO患者眶成纤维细胞OF分化重构进程的调节作用；②TAO模型小鼠的制作；③消瘿合剂含药血清对TAO模型小鼠T淋巴细胞活化，CD4/CD8表达比例的调控作用。以此来验证消瘿合剂通过抑制眶成纤维细胞、T淋巴细胞活化来抑制TAO炎症反应。结果显示，第一部分，对体外培养的眶成纤维细胞，消瘿合剂药物血清具有与PPARγ抑制剂相类似的对体外培养的眶成纤维细胞增殖有抑制作用，但不直接抑制OF增殖，而是通过抑制PPARγ分泌来抑制OF分化。且对OF分泌IGF-1也有抑制作用。第二部分通过质粒腺病毒转染技术制作TAO模型小鼠，经预实验、正式开展造模，积累了造模经验，造模成功率达40%。第三部分，开始针对模型小鼠及对照组用药，显示消瘿合剂能够有效减少模型组小鼠眼眶球后结缔组织充血及T淋巴细胞活化，抑制TAO免疫反应的发生。为消瘿合剂在TAO防治中的临床应用及推广提供科学依据。