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ENGLISH ABSTRACT
IL-17A和IFN-γ对甲状腺相关眼病患者CD34 -眼眶成纤维细胞纤维化的作用
鲁奕
黄雅琢
李寅炜
吴钰
方思捷
孙静
周慧芳
作者及单位信息
·
DOI： 10.3760/cma.j.cn115989-20200719-00513
Effects of IL-17A and IFN-γ on the fibrosis of CD34 - orbital fibroblasts in Graves orbitopathy
Lu Yi
Huang Yazhuo
Li Yinwei
Wu Yu
Fang Sijie
Sun Jing
Zhou Huifang
Authors Info & Affiliations
Lu Yi
Department of Ophthalmology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China
Huang Yazhuo
Department of Ophthalmology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China
Li Yinwei
Department of Ophthalmology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China
Wu Yu
Department of Ophthalmology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China
Fang Sijie
Department of Ophthalmology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China
Sun Jing
Department of Ophthalmology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China
Zhou Huifang
Department of Ophthalmology, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China
·
DOI： 10.3760/cma.j.cn115989-20200719-00513
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摘要

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CD34 -眼眶成纤维细胞(OFs)的作用，探讨GO的发病机制。

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CD34 - 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Western blot法检测各组眼眶脂肪结缔组织和泪腺组织来源的CD34 - 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。

结果在眼眶脂肪结缔组织来源的CD34 - 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 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 P<0.05)。在泪腺组织来源的CD34 - 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 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 P<0.05)。

结论高质量浓度IL-17A处理可促进TGF-β诱导的CD34 - OFs纤维化，而高质量浓度的IFN-γ则抑制TGF-β诱导的CD34 - OFs纤维化。

Graves眼病;成纤维细胞;转化生长因子-β;白细胞介素-17A;γ干扰素;纤维化
ABSTRACT

ObjectiveTo observe the effects of interleukin-17A (IL-17A) and interferon-γ (IFN-γ) on CD34 - orbital fibroblasts (OFs) in patients with Graves ophthalmopathy (GO), and to explore the pathogenesis of GO.

MethodsOrbital 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.CD34 - 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 CD34 - OFs derived from both orbital adipose connective tissue and lacrimal gland tissue were detected by Western blot.This study protocol was approved by an Ethic Committee of Shanghai Ninth People's Hospital (SH9H-2020-TK195-1) and written informed consent was obtained from each patient.

ResultsFor CD34 - 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 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 at P<0.05). For CD34 - 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 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 at P<0.05).

ConclusionsHigh level of IL-17A can promote the fibrosis of TGF-β-induced CD34 - OFs, and high level of IFN-γ inhibits the fibrosis of TGF-β-induced CD34 - OFs.

Graves ophthalmopathy;Fibroblast;Transforming growth factor-β;Interleukin-17A;Interferon-γ;Fibrosis
Zhou Huifang, Email: mocdef.3ab61gnafzzgnaf
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引用本文

鲁奕,黄雅琢,李寅炜,等. IL-17A和IFN-γ对甲状腺相关眼病患者CD34 -眼眶成纤维细胞纤维化的作用 [J]. 中华实验眼科杂志,2020,38(11)：923-928.

DOI:10.3760/cma.j.cn115989-20200719-00513

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甲状腺相关眼病(Graves ophthalmopathy，GO)是一种由多因素导致的自身免疫性眼眶病，发病率高，是Graves病最重要和常见的甲状腺外表现 [ 1 ]。近年来，眼外肌纤维化研究逐渐引起关注，为探讨GO的发病机制开辟了新的方向；而眼眶成纤维细胞(orbital fibroblasts，OFs)激活导致的眼外肌和眼眶脂肪纤维化被认为是GO发生和发展的关键环节 [ 2 ]。研究证实，OFs是GO自身免疫反应的效应细胞，其表面存在甲状腺刺激激素受体(thyroid stimulating hormone receptor，TSHR)及胰岛素样生长因子-1受体的自身抗原，与GO的发病密切相关 [ 3 ]。GO的主要病理改变为眼眶组织的免疫炎症反应，疾病的活动期主要表现为眼睑、结膜的充血水肿，静止期则出现眶内组织的纤维化和脂肪化，导致眼球运动障碍、斜视和复视。GO患者眼眶脂肪结缔组织中分别存在着CD34 ＋OFs和CD34 - OFs，这2类OFs在眼眶纤维化的发生和发展中起着不同的作用，其中GO活动期以CD34 ＋OFs的作用为主，静止期则以CD34 - OFs的作用为主 [ 4 ]。传统观点认为，B细胞、CD4 ＋Th1细胞和CD4 ＋Th2细胞均参与GO眼眶炎症反应 [ 5 ]，能较好地解释眼眶水肿的机制，但无法解释静止期GO发生纤维化和脂肪化病变的原因。最新研究显示，GO患者球后组织中存在一群高表达白细胞介素(interleukin，IL)-23受体的Th17细胞，可同时分泌γ干扰素(interferon-γ，IFN-γ)和IL-17A等细胞因子 [ 6 ]。本研究探讨IFN-γ和IL-17A在静止期GO眼眶组织发生纤维化改变中的作用。
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参考文献
[1]
Smith TJ , Hegedüs L ．Graves' Disease[J]．N Engl J Med， 2016，375(16):1552-1565. DOI： 10.1056/NEJMra1510030 .
返回引文位置Google Scholar
百度学术
万方数据
[2]
Fang S , Huang Y , Wang S ，et al. IL-17A exacerbates fibrosis by promoting the proinflammatory and profibrotic function of orbital fibroblasts in TAO[J]．J Clin Endocrinol Metab， 2016，101(8):2955-2965. DOI： 10.1210/jc.2016-1882 .
返回引文位置Google Scholar
百度学术
万方数据
[3]
Taylor PN , Zhang L , Lee R ，et al. New insights into the pathogenesis and nonsurgical management of Graves orbitopathy[J]．Nat Rev Endocrinol， 2020，16(2):104-116. 10.1038/s41574-019-0305-4 .
返回引文位置Google Scholar
百度学术
万方数据
[4]
Smith TJ . Potential roles of CD34＋ fibrocytes masquerading as orbital fibroblasts in thyroid-associated ophthalmopathy[J]．J Clin Endocrinol Metab， 2019，104(2):581-594. DOI： 10.1210/jc.2018-01493 .
返回引文位置Google Scholar
百度学术
万方数据
[5]
Smith TJ . TSH-receptor-expressing fibrocytes and thyroid-associated ophthalmopathy[J]．Nat Rev Endocrinol， 2015，11(3):171-181. DOI： 10.1038/nrendo.2014.226 .
返回引文位置Google Scholar
百度学术
万方数据
[6]
Huang Y , Fang S , Li D ，et al. The involvement of T cell pathogenesis in thyroid-associated ophthalmopathy[J]．Eye (Lond)， 2019，33(2):176-182. DOI： 10.1038/s41433-018-0279-9 .
返回引文位置Google Scholar
百度学术
万方数据
[7]
Bahn RS . Current insights into the pathogenesis of Graves' ophthalmopathy[J]．Horm Metab Res， 2015，47(10):773-778. DOI： 10.1055/s-0035-1555762 .
返回引文位置Google Scholar
百度学术
万方数据
[8]
Dominici M , Le Blanc K , Mueller I ，et al. Minimal criteria for defining multipotent mesenchymal stromal cells.The International Society for Cellular Therapy position statement[J]．Cytotherapy， 2006，8(4):315-317. DOI： 10.1080/14653240600855905 .
返回引文位置Google Scholar
百度学术
万方数据
[9]
Fernando R , Atkins SJ , Smith TJ . Slit2 may underlie divergent induction by thyrotropin of Il-23 and Il-12 in human fibrocytes[J]．J Immunol， 2020，204(7):1724-1735. DOI： 10.4049/jimmunol.1900434 .
返回引文位置Google Scholar
百度学术
万方数据
[10]
Fernando R , Grisolia A , Lu Y ，et al. Slit2 modulates the inflammatory phenotype of orbit-infiltrating fibrocytes in Graves' disease[J]．J Immunol， 2018，200(12):3942-3949. DOI： 10.4049/jimmunol.1800259 .
返回引文位置Google Scholar
百度学术
万方数据
[11]
Isono F , Fujita-Sato S , Ito S ．Inhibiting RORγt/Th17 axis for autoimmune disorders[J]．Drug Discov Today， 2014，19(8):1205-1211. DOI： 10.1016/j.drudis.2014.04.012 .
返回引文位置Google Scholar
百度学术
万方数据
[12]
Shabgah AG , Fattahi E , Shahneh FZ . Interleukin-17 in human inflammatory diseases[J]．Postepy Dermatol Alergol， 2014，31(4):256-261. DOI： 10.5114/pdia.2014.40954 .
返回引文位置Google Scholar
百度学术
万方数据
[13]
Fang S , Huang Y , Liu X ，et al. Interaction between CCR6＋ Th17 cells and CD34＋ fibrocytes promotes i nflammation：implications in Graves' orbitopathy in Chinese population [J]．Invest Ophthalmol Vis Sci， 2018，59(6):2604-2614. DOI： 10.1167/iovs.18-24008 .
返回引文位置Google Scholar
百度学术
万方数据
[14]
Fang S , Huang Y , Wang N ，et al. Insights into local orbital immunity：evidence for the involvement of the Th17 cell pathway in thyroid-associated ophthalmopathy[J]．J Clin Endocrinol Metab， 2019，104(5):1697-1711. DOI： 10.1210/jc.2018-01626 .
返回引文位置Google Scholar
百度学术
万方数据
[15]
Huh HD , Kim JH , Kim SJ ，et al. The change of lacrimal gland volume in Korean patients with thyroid-associated ophthalmopathy[J]．Korean J Ophthalmol， 2016，30(5):319-325. DOI： 10.3341/kjo.2016.30.5.319 .
返回引文位置Google Scholar
百度学术
万方数据
[16]
Eckstein AK , Finkenrath A , Heiligenhaus A ，et al. Dry eye syndrome in thyroid-associated ophthalmopathy：lacrimal expression of TSH receptor suggests involvement of TSHR-specific autoantibodies[J]．Acta Ophthalmol Scand， 2004，82(3Pt 1):291-297. DOI： 10.1111/j.1395-3907.2004.00268.x .
返回引文位置Google Scholar
百度学术
万方数据
[17]
Huang D , Luo Q , Yang H ，et al. Changes of lacrimal gland and tear inflammatory cytokines in thyroid-associated ophthalmopathy[J]．Invest Ophthalmol Vis Sci， 2014，55(8):4935-4943. DOI： 10.1167/iovs.13-13704 .
返回引文位置Google Scholar
百度学术
万方数据
[18]
Fang S , Huang Y , Zhong S ，et al. Regulation of orbital fibrosis and adipogenesis by pathogenic Th17 cells in Graves orbitopathy[J]．J Clin Endocrinol Metab， 2017，102(11):4273-4283. DOI： 10.1210/jc.2017-01349 .
返回引文位置Google Scholar
百度学术
万方数据
[19]
Poosti F , Bansal R , Yazdani S ，et al. Selective delivery of IFN-γ to renal interstitial myofibroblasts：a novel strategy for the treatment of renal fibrosis[J]．FASEB J， 2015，29(3):1029-1042. DOI： 10.1096/fj.14-258459 .
返回引文位置Google Scholar
百度学术
万方数据
备注信息
A
周慧芳，Email： mocdef.3ab61gnafzzgnaf
B
所有作者均声明不存在利益冲突
C
国家重点研发计划项目 (2018YFC1106100)
国家自然科学基金面上项目 (81770960)
国家自然科学基金国际（地区）合作与交流项目 (81761168037)
上海市科学技术委员会基金项目 (17DZ2260100)
上海市教委"高水平地方高校建设创新团队"眼肿瘤与眼眶修复协同创新团队 (SSMV-ZDCX20180401)
上海交通大学医学院附属第九人民医院院级基金项目 (JYHL2019MS04)
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