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目的:观察白细胞介素-17A(IL-17A)与γ干扰素(IFN-γ)对Graves眼病(GO)患者CD34n -眼眶成纤维细胞(OFs)的作用，探讨GO的发病机制。n 方法:收集5例GO患者眼眶减压术中获取的眼眶脂肪结缔组织以及泪腺组织，采用组织块消化法进行培养和传代。采用免疫磁珠富集其中的CD34n - OFs。将培养的OFs分为转化生长因子-β(TGF-β)组、TGF-β＋10 ng/ml IL-17A组和TGF-β＋100 ng/ml IL-17A组、TGF-β＋1 ng/ml IFN-γ组和TGF-β＋5 ng/ml IFN-γ组，在各组细胞培养液中均添加5 ng/ml TGF-β以诱导细胞发生纤维化，按照分组方法分别加入不同质量浓度的IL-17A或IFN-γ。采用Weste blot法检测各组眼眶脂肪结缔组织和泪腺组织来源的CD34n - OFs中纤连蛋白、Ⅰ型胶原、ɑ-平滑肌肌动蛋白(ɑ-SMA)和组织金属基质蛋白酶抑制剂1(TIMP-1)等纤维化相关蛋白表达。n 结果:在眼眶脂肪结缔组织来源的CD34n - OFs中，TGF-β＋100 ng/ml IL-17A组细胞中纤连蛋白、Ⅰ型胶原、α-SMA和TIMP-1蛋白相对表达量明显高于TGF-β组和TGF-β＋10 ng/ml IL-17A组，差异均有统计学意义(均n P<0.05)；TGF-β＋5 ng/ml IFN-γ组细胞中纤连蛋白、Ⅰ型胶原和α-SMA相对表达量均明显低于TGF-β组和TGF-β＋1 ng/ml IFN-γ组，差异均有统计学意义(均n P<0.05)。在泪腺组织来源的CD34n - OFs中，TGF-β＋100 ng/ml IL-17A组细胞中纤连蛋白、Ⅰ型胶原、α-SMA和TIMP-1蛋白相对表达量明显高于TGF-β组和TGF-β＋10 ng/ml IL-17A组，差异均有统计学意义(均n P<0.05)；TGF-β＋1 ng/ml IFN-γ组细胞中纤连蛋白、Ⅰ型胶原和TIMP-1蛋白相对表达量明显高于TGF-β组和TGF-β＋5 ng/ml IFN-γ组，而α-SMA蛋白相对表达量明显高于TGF-β＋5 ng/ml IFN-γ组，差异均有统计学意义(均n P<0.05)。n 结论:高质量浓度IL-17A处理可促进TGF-β诱导的CD34n - OFs纤维化，而高质量浓度的IFN-γ则抑制TGF-β诱导的CD34n - OFs纤维化。n ","Objective:To observe the effects of interleukin-17A (IL-17A) and interferon-γ (IFN-γ) on CD34n - orbital fibroblasts (OFs) in patients with Graves ophthalmopathy (GO), and to explore the pathogenesis of GO.n Methods:Orbital adipose connective tissue and lacrimal gland tissue of 5 patients with GO were collected during orbital decompression surgery.These tissue was cultured by tissue explant culture method.CD34n - OFs were enriched by immunomagnetic beads after passage and expansion.The cultured cells were divided into transforming growth factor-β (TGF-β) group, TGF-β+ 10 ng/ml IL-17A group, TGF-β+ 100 ng/ml IL-17A group, TGF-β+ 1 ng/ml IFN-γ group and TGF-β+ 5 ng/ml IFN-γ group.The fibrosis of the cells was induced with 5 ng/ml TGF-β and then was treated with different concentrations of IL-17A or IFN-γ according to grouping.The expression of fibronectin, collagen type Ⅰ, α-smooth muscle actin (α-SMA) and tissue metalloproteinase inhibitor-1 (TIMP-1) in CD34n - OFs derived from both orbital adipose connective tissue and lacrimal gland tissue were detected by Weste blot.This study protocol was approved by an Ethic Committee of Shanghai Ninth Peoples Hospital (SH9H-2020-TK195-1) and written informed consent was obtained from each patient.n Results:For CD34n - OFs derived from orbital adipose connective tissue, the relative expressions of fibronectin, type Ⅰ collagen, α-SMA and TIMP-1 protein were significantly higher in the TGF-β+ 100 ng/ml IL-17A group than those in the TGF-β group and TGF-β+ 10 ng/ml IL-17A group (all at n P<0.05); the relative expressions of fibronectin, type Ⅰ collagen and α-SMA in the cells in the TGF-β+ 5 ng/ml IFN-γ group were significantly lower than those in the TGF-β group and the TGF-β+ 1 ng/ml IFN-γ group (all atn P<0.05). For CD34n - OFs derived from lacrimal gland, the relative expressions of fibronectin, type Ⅰ collagen, α-SMA and TIMP-1 protein in the TGF-β+ 100 ng/ml IL-17A group were significantly higher than those in the TGF-β group and the TGF-β+ 10 ng/ml IL-17A group (all at n P<0.05); the relative expressions of fibronectin, type Ⅰ collagen and TIMP-1 protein in the TGF-β+ 1 ng/ml IFN-γ group were significantly higher than those in the TGF-β group and TGF-β+ 5 ng/ml IFN-γ group, and the relative expression of α-SMA protein was significantly higher than that in the TGF-β+ 5 ng/ml IFN-γ group (all atn P<0.05).n Conclusions:High level of IL-17A can promote the fibrosis of TGF-β-induced CD34n - OFs, and high level of IFN-γ inhibits the fibrosis of TGF-β-induced CD34n - OFs.n