Department of Immunology, Erasmus MC, University Medical Center, Rotterdam, The Netherlands.
Department of Ophthalmology, Erasmus MC, University Medical Center, Rotterdam, The Netherlands The Rotterdam Eye Hospital, Rotterdam, The Netherlands.
Invest Ophthalmol Vis Sci. 2014 Jul 11;55(7):4659-66. doi: 10.1167/iovs.14-14818.
To determine the role of thrombin in the development of proliferative vitreoretinopathy (PVR).
Vitreous was collected from patients undergoing a vitrectomy (macular holes and puckers, n = 11 [controls]; retinal detachment without PVR development following vitrectomy, n = 15 [RRD1]; retinal detachment with PVR development within 6 months after vitrectomy, n = 11 [RRD2]; and established PVR, n = 14 [PVR]). Thrombin activity in vitreous was determined using a thrombin-specific chromogenic substrate. ARPE-19 cells were stimulated with 8× diluted vitreous samples in the presence and absence of hirudin. The samples were analyzed at t = 0 and t = 24 hours for the presence of 27 cytokines/chemokines and growth factors using a multiplex approach. In comparable studies, ARPE-19 cells were stimulated for 2 hours, and mRNA expression levels for CCL2, CXCL8, GMCSF, IL6, and PDGFB were determined by real-time quantitative (RQ)-PCR.
Thrombin activity was significantly (P < 0.05) higher in vitreous of the PVR group compared to the other groups. Proliferative vitreoretinopathy vitreous stimulated the production of chemokine (C-C motif) ligand (CCL)2, chemokine (C-X-C motif) ligand (CXCL)8, granulocyte-macrophage colony-stimulating factor (GM-CSF), IL-6, and platelet-derived growth factor (PDGF)-BB by ARPE-19 to significantly (P < 0.05) higher levels than vitreous from the RRD1 and RRD2 groups. These effects of PVR vitreous were significantly (P < 0.05) reduced by hirudin. These data were confirmed by mRNA studies.
Thrombin activity is increased in vitreous of patients with established PVR and is involved in the activation of proinflammatory and profibrotic pathways in RPE cells. Inhibition of thrombin activity may therefore represent a potential treatment option for proliferative vitreoretinopathy.
确定凝血酶在增生性玻璃体视网膜病变（PVR）发展中的作用。
收集行玻璃体切除术患者的玻璃体（黄斑裂孔和皱褶，n = 11[对照组]；玻璃体切除术后无 PVR 发展的视网膜脱离，n = 15[RRD1]；玻璃体切除术后 6 个月内发生 PVR 发展的视网膜脱离，n = 11[RRD2]；以及已建立的 PVR，n = 14[PVR]）。使用凝血酶特异性显色底物测定玻璃体中的凝血酶活性。用 8 倍稀释的玻璃体样本刺激 ARPE-19 细胞，并在存在和不存在水蛭素的情况下进行刺激。在 t = 0 和 t = 24 小时时，使用多重方法分析样本中 27 种细胞因子/趋化因子和生长因子的存在情况。在可比的研究中，用凝血酶刺激 ARPE-19 细胞 2 小时，并通过实时定量（RQ）-PCR 测定 CCL2、CXCL8、GMCSF、IL6 和 PDGFB 的 mRNA 表达水平。
与其他组相比，PVR 组玻璃体中的凝血酶活性显著升高（P < 0.05）。增生性玻璃体视网膜病变玻璃体刺激 ARPE-19 产生趋化因子（C-C 基序）配体（CCL）2、趋化因子（C-X-C 基序）配体（CXCL）8、粒细胞-巨噬细胞集落刺激因子（GM-CSF）、白细胞介素 6（IL-6）和血小板衍生生长因子（PDGF）-BB 的产生水平显著升高（P < 0.05）比 RRD1 和 RRD2 组的玻璃体高。水蛭素可显著降低（P < 0.05）PVR 玻璃体的这些作用。mRNA 研究证实了这些数据。
在已建立的 PVR 患者的玻璃体中，凝血酶活性增加，并参与 RPE 细胞中促炎和促纤维化途径的激活。因此，抑制凝血酶活性可能是治疗增生性玻璃体视网膜病变的一种潜在治疗选择。
