Evidence for Enhanced Thy-1 (CD90) Expression in Orbital Fibroblasts of Patients with Graves' Ophthalmopathy
Evidence for Enhanced Thy-1 (CD90) Expression in Orbital Fibroblasts of Patients with Graves' Ophthalmopathy
复制标题
DOI：
10.1089/thy.2008.0255
复制
发表时间：
2008-12-01
期刊：
影响因子：
6.6
通讯作者：
Bahn, Rebecca S.
中科院分区：
文献类型：
--
作者：
Khoo, Teck Kim;Coenen, Michael J.;Bahn, Rebecca S.
背景：THY-1是一种表面蛋白，它定义了功能上不同的成纤维细胞亚群，那些缺乏抗原的成纤维细胞能够成脂。由于脂肪细胞发育增加是Graves眼病(GO)眼眶的特征，我们希望比较GO患者和正常人眼眶成纤维细胞中Thy-1的基线表达，并确定过氧化物酶增殖物激活物-伽玛结扎后脂肪生成是否会影响该蛋白的水平。方法：眼眶脂肪/结缔组织标本来自接受眼眶减压手术的重度GO患者(n=9)和正常人(n=9)。用RT-PCR、定量免疫荧光染色和使用Thy-1鼠抗人CD90/Thy-1单抗的流式细胞仪检测组织标本和眼眶成纤维细胞培养物中THY-1mRNA和蛋白的水平。结果：未培养的GO结缔组织和脂肪组织中Thy-1mRNA和蛋白的表达水平(3.8倍；0.835+/-0.116相对表达)高于正常(0.22+/-0.062；p=0.002)和培养的GO患者眼眶成纤维细胞(3.3倍；9.28+/-1.82相对表达)；P=0.013)。脂肪细胞分化对Thy-1表达无影响。流式细胞仪和免疫荧光染色显示GO中Thy-1阳性细胞数量(平均77.9±4.09%；范围66.5~84.8%)高于正常成纤维细胞培养(66.8±1.6%；63.3~71.0%；p=0.046)，且阳性细胞表达水平较高。结论：GO眼眶组织和培养物中Thy-1表达增加可能是眼眶疾病过程的结果，反映了Thy-1阳性细胞数量的增加和这些细胞的高表达。脂肪生成本身似乎不会影响Thy-1的表达。这种蛋白在GO中的表达增加可能代表着对细胞损伤的适应性反应，从而有效地限制了眼眶脂肪/结缔组织内的疾病进展。
Background: Thy-1 is a surface protein that defines functionally distinct subpopulations of fibroblasts, with those lacking the antigen being capable of adipogenesis. Because increased fat cell development is a hallmark of the orbit in Graves' ophthalmopathy (GO), we wished to compare baseline Thy-1 expression in orbital fibroblasts from GO patients and normal individuals, and determine whether levels of the protein might be impacted by adipogenesis following peroxisome proliferator activator-gamma ligation.Methods: Orbital adipose/connective tissue specimens were obtained from euthyroid patients undergoing orbital decompression surgery for severe GO (n = 9) and from normal individuals (n = 9). Thy-1 mRNA and protein levels were assessed in tissue specimens and in orbital fibroblast cultures at baseline using RT-PCR, quantitative immunofluorescent staining, and flow cytometry using a specific Thy-1 mouse anti-human CD90/Thy-1 monoclonal antibody. In addition, some orbital fibroblast cultures were treated with rosiglitazone (1 mu L/mL; 2 nM) or control for 10 days in culture.Results: We found that Thy-1 mRNA and protein expression was higher in uncultured GO connective/adipose tissue specimens (3.8-fold; 0.835 +/- 0.116 relative expression) compared with normal (0.22 +/- 0.062; p = 0.002) and in cultured orbital fibroblasts from GO patients (3.3-fold; 9.28 +/- 1.82 relative expression) compared with normal cultures (2.80 +/- 0.42; p = 0.013). Adipocyte differentiation had no effect on Thy-1 expression. Flow cytometry and immunofluorescent staining showed increased numbers of Thy-1-positive cells in the GO (mean 77.9 + 4.09%; range 66.5-84.8%) compared with the normal fibroblast cultures (66.8 + 1.6%; range 63.3-71.0% positive; p = 0.046), as well as higher levels of expression on the positive cells.Conclusions: Increased Thy-1 expression in GO orbital tissues and cultures is likely a consequence of the orbital disease process, reflecting both the presence of increased numbers of Thy-1-positive cells and higher expression on those cells. Adipogenesis itself does not appear to impact Thy-1 expression. Increased expression of this protein in GO could represent an adaptive response to cell injury, in effect limiting disease progression within the orbital adipose/connective tissues.