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  • 刁嘉乐,魏锐利*.穿透素3对转化生长因子β1诱导的眼眶成纤维细胞纤维化的影响[J].第二军医大学学报,2020,41(4):424-428    [点击复制]
  • DIAO Jia-le,WEI Rui-li*.Effect of pentraxin 3 on transforming growth factor β1-induced fibrosis in orbital fibroblasts[J].Acad J Sec Mil Med Univ,2020,41(4):424-428   [点击复制]
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穿透素3对转化生长因子β1诱导的眼眶成纤维细胞纤维化的影响
刁嘉乐,魏锐利*
0
(海军军医大学(第二军医大学)长征医院眼科, 上海 200003
*通信作者)
摘要:
目的 探索穿透素3(PTX3)对TGF-β诱导的眼眶成纤维细胞(OF)纤维化的作用。方法 以体外培养的甲状腺相关眼病(TAO)患者来源及健康来源OF为研究对象,在培养液中加入人重组PTX3(rhPTX3)蛋白(1 μg/mL)及重组TGF-β1(10 ng/mL)进行刺激,刺激完成后选择特异性抗体标记细胞中α-平滑肌肌动蛋白(α-SMA),通过免疫荧光显微镜观察OF的纤维化程度变化。给予细胞相同的刺激,刺激后通过qRT-PCR检测细胞中α-SMA、Ⅰ型胶原蛋白α1(ColIα1)、IL-6 mRNA表达量的变化。结果 TGF-β1可诱导两组OF中α-SMA蛋白表达量增加,细胞呈现纤维化改变,而rhPTX3对该效应未见明显抑制作用。TAO患者来源OF的纤维化效应较健康来源OF无明显差异。TGF-β1可诱导OF纤维化相关蛋白(α-SMA、ColIα1、IL-6)mRNA表达上调。rhPTX3对OF中α-SMA mRNA的表达无明显诱导作用,但共刺激时可增强TGF-β1对α-SMA mRNA的上调效应。rhPTX3可上调ColIα1IL-6 mRNA的表达,共刺激时可增强TGF-β1对IL-6 mRNA的诱导上调,但对ColIα1 mRNA的上调未见明显影响。结论 rhPTX3对TGF-β1诱导的OF纤维化过程具有潜在的促进作用,说明PTX3可能参与了TAO患者后期视物重影的发病过程,并发挥了正向作用。
关键词:  穿透素3  甲状腺相关眼病  眼眶成纤维细胞  转化生长因子β  纤维化
DOI:10.16781/j.0258-879x.2020.04.0424
投稿时间:2020-03-09修订日期:2020-04-07
基金项目:国家自然科学基金(81770959),国家科技重大专项(2017ZX09304010).
Effect of pentraxin 3 on transforming growth factor β1-induced fibrosis in orbital fibroblasts
DIAO Jia-le,WEI Rui-li*
(Department of Ophthalmology, Changzheng Hospital, Naval Medical University (Second Military Medical University), Shanghai 200003, China
*Corresponding author)
Abstract:
Objective To explore the role of pentraxin 3 (PTX3) in transforming growth factor β1 (TGF-β1)-induced fibrosis in orbital fibroblasts (OFs). Methods OFs from thyroid-associated ophthalmopathy (TAO) patients and healthy donors were cultured in vitro. The human recombinant PTX3 (rhPTX3) (1 μg/mL) and human recombinant TGF-β1 (10 ng/mL) were added to the culture medium for stimulation. After stimulation, specific antibody was selected to label the anti-α-smooth muscle actin (α-SMA), and the fibrosis of OFs was observed by fluorescence microscope. The mRNA expression levels of α-SMA, collagen typeⅠα1 (ColIα1) and interleukin 6 (IL-6) were detected by quantitative real-time polymerase chain reaction after the same stimulation. Results TGF-β1 induced the expression of α-SMA protein in the two groups of OFs, and the cells showed fibrotic changes, while the rhPTX3 had no significant inhibition. There was no significant difference in the fibrotic effect of OFs between the patients with TAO and healthy donors. The mRNA expression levels of fibrotic protein (α-SMA, ColIα1, and IL-6) were upregulated by TGF-β1 in OFs. The rhPTX3 had no significant induction for the expression of α-SMA mRNA in OFs, but co-stimulation could significantly enhance the upregulation of α-SMA. The rhPTX could upregulate the mRNA expression of ColIα1 and IL-6. Co-stimulation could enhance the induction of upregulation of IL-6 by TGF-β1, but had no significant effect on the upregulation of ColIα1. Conclusion The rhPTX3 has a potential role in promoting TGF-β1-induced fibrosis in OFs, which indicates that PTX3 may be involved in the later stage of the pathogenesis of diplopia in TAO patients and play a positive role.
Key words:  pentraxin 3  thyroid associated ophthalmopathy  orbital fibroblasts  transforming growth factor β  fibrosis