Han Han, Zhao Xiao, Liao Mengyu, Song Yinting, You Caiyun, Dong Xue, Yang Xueli, Wang Xiaohong, Huang Bo, Du Mei, Yan Hua
Department of Ophthalmology, Tianjin Medical University General Hospital, Tianjin, China.
Laboratory of Molecular Ophthalmology, Department of Pharmacology and Tianjin Key Laboratory of Inflammation Biology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin, China.
Invest Ophthalmol Vis Sci. 2021 Jul 1;62(9):29. doi: 10.1167/iovs.62.9.29.
Uncontrolled coagulation reactions contribute to pathological fibroproliferation in several organs, and yet their role in proliferative vitreoretinopathy (PVR) remains to be elucidated. In this study, we evaluated the profibrotic effects of FXa in RPE cells and in a mouse model of PVR.
FXa levels in the eyes of traumatic PVR patients and rabbit models of mechanical ocular trauma was measured by ELISA and immunohistochemistry. FXa-induced RPE EMT was assessed by examining cell proliferation, migration, tight junction changes, and expression of fibrotic markers. For in vivo study, FXa was injected into dispase-injured eyes, then intraocular fibrosis was evaluated by histological analysis and Western blotting. The therapeutic effect of FXa inhibitor was also examined in PVR mouse models.
Vitreous FXa were higher in patients with traumatic PVR compared to patients with macular hole. Moreover, expressions of FXa and PAR1 were found in the epiretinal membranes from traumatic PVR patients. Vitreous FXa were markedly increased after mechanical ocular trauma in rabbits. In vitro, FXa stimulated RPE EMT characterized as ZO-1 disruption, compromised cell polarity, and increased fibronectin expressions. Co-injection of FXa and dispase in mice induced more severely damaged retinal structures, and increased α-SMA expressions than FXa or dispase treatment alone. Oral FXa or thrombin inhibitors significantly blocked intraocular fibrosis in PVR mouse models. FXa promoted phospho-activation of p38 in ARPE19 cells, which was dependent on PAR1. Moreover, TGF-βR inhibitor also significantly alleviated FXa-induced intraocular fibrosis in mice.
FXa promotes intraocular fibrosis in mice via mechanisms involving RPE activation.
失控的凝血反应会导致多个器官发生病理性纤维增生，但其在增殖性玻璃体视网膜病变（PVR）中的作用仍有待阐明。在本研究中，我们评估了因子Xa（FXa）在视网膜色素上皮（RPE）细胞和PVR小鼠模型中的促纤维化作用。
通过酶联免疫吸附测定（ELISA）和免疫组织化学法检测外伤性PVR患者及机械性眼外伤兔模型眼中的FXa水平。通过检测细胞增殖、迁移、紧密连接变化及纤维化标志物的表达来评估FXa诱导的RPE上皮-间质转化（EMT）。在体内研究中，将FXa注入经dispase损伤的眼中，然后通过组织学分析和蛋白质免疫印迹法评估眼内纤维化情况。还在PVR小鼠模型中检测了FXa抑制剂的治疗效果。
与黄斑裂孔患者相比，外伤性PVR患者玻璃体内的FXa水平更高。此外，在外伤性PVR患者的视网膜前膜中发现了FXa和蛋白酶激活受体1（PAR1）的表达。兔机械性眼外伤后玻璃体内的FXa明显增加。在体外，FXa刺激RPE发生EMT，表现为紧密连接蛋白1（ZO-1）破坏、细胞极性受损及纤连蛋白表达增加。在小鼠中联合注射FXa和dispase比单独注射FXa或dispase诱导的视网膜结构损伤更严重，α-平滑肌肌动蛋白（α-SMA）表达增加。口服FXa或凝血酶抑制剂可显著阻断PVR小鼠模型中的眼内纤维化。FXa促进ARPE19细胞中p38的磷酸化激活，这依赖于PAR1。此外，转化生长因子-β受体（TGF-βR）抑制剂也可显著减轻FXa诱导的小鼠眼内纤维化。
FXa通过涉及RPE激活的机制促进小鼠眼内纤维化。
