Effects of IL-17A and IFN-γ on the fibrosis of CD34- orbital fibroblasts in Graves orbitopathy

Authors: Lu Yi,  Huang Yazhuo,  Li Yinwei,  Wu Yu,  Fang Sijie,  Sun Jing,  Zhou Huifang
DOI: 10.3760/cma.j.cn115989-20200719-00513
Published 2020-11-10
Cite as Chin J Exp Ophthalmol, 2020,38(11): 923-928.

Abstract

Objective

To 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.

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.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.

Results

For 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).

Conclusions

High 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.

Key words:

Graves ophthalmopathy; Fibroblast; Transforming growth factor-β; Interleukin-17A; Interferon-γ; Fibrosis

Contributor Information

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
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Updated: November 19, 2020 — 3:56 am