胰岛素样生长因子-1对甲状腺相关眼病眼眶成纤维细胞的促生长作用  

Promoting effects of insulin-like growth factor-1 on proliferation of orbital fibroblasts derived from thyroid associated ophthalmopathy

作　　者：代佳灵 何为民[1] 罗梦绮 

机构地区：[1]四川大学华西医院眼科,成都610000

出　　处：《中华实验眼科杂志》2017年第9期805-810,共6页Chinese Journal Of Experimental Ophthalmology

基　　金：四川省科技计划项目（2013SZ0029）;成都市惠民科技项目（2014-HM01-001420SF）

摘　　要：背景甲状腺相关眼病（TAO）是一种自身免疫性疾病，目前对其发病机制的研究主要集中在共同抗原上。胰岛素样生长因子-1（IGF．1）功能的发挥需要IGF-1受体（IGF-1R）的参与，IGF-1在促甲状腺激素受体（TSHR）的信号传导通路中也起着重要作用。目的探讨IGF-1对TAO患者眼眶成纤维细胞（OFs）增生及IGF-1R、TSHR表达的影响，探讨IGF-1在TAO发病机制中的作用。方法收集2016年3-6月于四川大学华西医院行TAO眼眶脂肪切除术取得的眶组织标本17例17份及正常人美容手术取得的眼眶组织4人4份，采用含体积分数5％胎牛血清的DMEM／F12培养液培养OFs。分别于培养液中加入不同质量浓度（O、50、100、125μg／L）的IGF-1，采用MTS比色法绘制OFs生长曲线，评估IGF-1对TAO和正常来源OFs增生的影响；采用流式细胞术测定不同质量浓度IGF-1对TAO和正常来源OFs中IGF-1R、TSHR表达水平的影响。结果TAO患者与正常人来源的OFs均生长良好，呈梭形，细胞质丰富，TAO患者与正常人来源的OFs形态学特点一致。培养的OFs波形蛋白呈阳性反应，结蛋白、S-100、肌红蛋白和角蛋白均呈阴性反应。不同质量浓度IGF-1作用后TAO组和正常组OFs增生均呈逐渐上升趋势，总体比较差异有统计学意义（F分组=219．639，P〈0．001；F质量浓度=17．752，P〈0．001），以100μg／LIGF-1的促进作用最强。0、50、100、125μg／LIGF-1作用后TAO组OFs中IGF-1R表达量分别为（0．0091±0．0087）％、（0．0953±0．0233）％、（0．0837±0．0227）％和（0．0709±0．0241）％，正常组OFs中IGF-1R表达量分别为（0．0023±0．0006）％、（0．0093±0．0012）％、（0．0073±0．0015）％和（0．0083±0．0012）％，其中50、100和125μg／LIGF-1作用后各组OFs中IGF-1R表达量均高于无添加IGF-1者，TAO组OFs细胞中IGF-1R的表达量均明显高于正常组，差异均有统计学意义（均P〈0．Background Thyroid associated ophthalmopathy （TAO） is an autoimmune disease. Current research on the pathogenesis focuses on common autoantigen. Insulin-like growth factor-1 receptor （IGF-1R） is necessary for the function of IGF-1 ,also IGF-1 plays an important role in signaling pathway of thyroid stimulating hormone receptor （TSHR）. Objective This study was to investigate the effects of IGF-1 on the proliferation, expression of IGF-1R and TSHR on cultured orbital fibroblasts （OFs） derived from TAO. Methods Human orbital tissue was obtained from 17 TAO patients who received orbital adipectomy and 4 normal controls who received cosmetic surgery in West China Hospital from March 2016 to June 2016. OFs were cultured by explant culture with DMEM/F12 containing 5% fetal bovine serum and identified by immunochemistry. The OFs were treated with different concentrations of IGF-1. IGF-1 at different concentrations （0,50,100,125μg/L） was added into the medium,respectively,and the proliferation of the cells （absorbancy） was detected by MTS. The percentages of IGF-1R and TSI-IR expressions in the ceils were assayed by flow cytometry. Results Cultured ceils appeared to be spindle-like in shape and grew well with abundant cytoplasm. The characteristics of the cells derived from TAO patients were consistant with normal ones. The cells showed the positive response for vimentin and absent respose for desmin,S-100,myoglobin and cytokeratin. The proliferative values of OFs were gradually elevated with the increase of IGF-1 dose in both TAO group and normal group （ Fsroup = 219. 639, P〈0. 001 ; Fconcentration= 17. 752, P〈0. 001 ） with the optimal effects in 100 μg/L IGF-1. The expression levels of IGF-1R in the OFs were （0. 009 1 ±0. 008 7）% , （0. 095 3±0. 023 3 ） % , （ 0. 083 7 ±0. 022 7 ） % and （ 0. 070 9 ± 0. 024 1 ） % in the TAO group, and those in the normal group were （0.0023± 0.0006）%,（0.0093±0.0012）%,（0.0073±0.0015）% and （0.0083±0.001 2）% after treatme

关 键 词：甲状腺相关眼病 眼眶成纤维细胞/代谢 胰岛素样生长因子-1/代谢 胰岛素样生长因子-1受体/代谢 促甲状腺激素受体/代谢 细胞增生/药物作用 细胞培养 

分 类 号：R581[医药卫生—内分泌] R771.3[医药卫生—内科学]