Zhang Ya-Ni, Dong Yan-Ling, Hao Wen-Pei, Bai Xiao-Fei, Qi Xia, Liu Ting, Sun Xiao-Tong, Wei Chao, Qi Xiao-Lin
Xi'an Children's Hospital, Xi'an 710004, Shaanxi Province, China.
Shandong Eye Institute, Shandong First Medical University & Shandong Academy of Medical Sciences, Qingdao 266071, Shandong Province, China.
Int J Ophthalmol. 2021 Nov 18;14(11):1660-1665. doi: 10.18240/ijo.2021.11.03. eCollection 2021.
To explore the expression of cGAS/STING signaling components in Mooren's ulcer (MU).
Samples were obtained from ten MU patients, and eight residual corneal-scleral rings of healthy donor corneas for controls. Human corneal epithelial cells (HCECs) were used to evaluate the effect of cGAS/STING signaling pathway. Immunohistochemistry (IHC) and Western blot were used to examine the expression of cGAS, STING, and phosphorylated interferon regulatory factor 3 (p-IRF3) in MU tissues. The expression of interferon-β (IFN-β) and interferon-stimulated genes (ISGs) was quantified by real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA).
The protein levels of cGAS and STING in MU samples were significantly elevated when compared with the healthy controls by Western blot and IHC. After stimulation with cGAMP, real-time PCR and ELISA showed a dramatic increase of IFN-β and ISGs (containing CXCL10, IFIT1, and IL-6) in HCECs. Moreover, HCECs treated with cGAMP was characterized by increased phosphorylation and more nuclear translocation of IRF3. Meanwhile, increased p-IRF3 was observed in MU samples IHC and Western blot.
The pronounced expression of cGAS/STING signaling components in the patients with MU and probably contribute to the onset and development of MU.
探讨cGAS/STING信号通路成分在蚕蚀性角膜溃疡(MU)中的表达情况。
收集10例MU患者的样本,并选取8个健康供体角膜的残余角膜巩膜环作为对照。用人角膜上皮细胞(HCECs)评估cGAS/STING信号通路的作用。采用免疫组织化学(IHC)和蛋白质印迹法检测MU组织中cGAS、STING和磷酸化干扰素调节因子3(p-IRF3)的表达。通过实时聚合酶链反应(PCR)和酶联免疫吸附测定(ELISA)对干扰素-β(IFN-β)和干扰素刺激基因(ISGs)的表达进行定量分析。
通过蛋白质印迹法和IHC检测发现,与健康对照相比,MU样本中cGAS和STING的蛋白水平显著升高。用2'3'-环鸟苷酸-腺苷酸(cGAMP)刺激后,实时PCR和ELISA结果显示HCECs中IFN-β和ISGs(包括CXCL10、IFIT1和IL-6)显著增加。此外,用cGAMP处理的HCECs表现为IRF3磷酸化增加和更多的核转位。同时,在MU样本的IHC和蛋白质印迹检测中观察到p-IRF3增加。
MU患者中cGAS/STING信号通路成分表达明显,可能参与了MU的发生和发展。