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Abstract：
Though the pathogenesis of choroidal neovascularization (CNV) is largely unknown in age-related macular degeneration (AMD), inflammasomes may contribute to CNV development and progression. To understand the role NLRP3 inflammasomes in CNV, we used Ccr2RFPCx3cr1GFP dual-reporter mice to characterize migration of Ccr2RFP positive monocytes and Cx3cr1GFP positive microglial cells into CNV lesions after laser-induced rupture of Bruch\'s membrane. MCC950 was used as NLRP3 inhibitor. Immunostaining was used to confirm localization of NLRP3 inflammasomes in the LCNV lesions. Confocal microscopy was used to image and quantify LCNV volumes. ELISA and qRT-PCR were used to confirm the activation of NLRP3 by monitoring the expression of IL-1β protein and mRNA in choroidal tissues from LCNV mice. In addition, NLRP3 (-/-) LCNV mice were used to investigate whether NLRP3 inflammasomes contribute to the development of LCNV lesions. We observed that RFP positive monocyte-derived macrophages and GFP positive microglia-derived macrophages, in addition to other cell types, were localized in LCNV lesions at day 7 post-laser injury. In addition, NLRP3 inflammasomes are associated with LCNV lesions. Inhibition of NLRP3 inflammasomes, using MCC950, caused an increased Ccr2RFP positive macrophages, Cx3cr1GFP positive microglia, and other cells resulting in an increase in total lesion size. NLRP3 (-/-) LCNV mice, showed significantly increased lesion size compared to age-matched controls. Inhibition of NLRP3, resulted in decreased IL-1β mRNA and protein expression in the choroidal tissues, suggesting that increased lesion size may not be directly related to IL-1β.
摘要：
尽管年龄相关性黄斑变性（AMD）中脉络膜新生血管（CNV）的发病机制在很大程度上是未知的，炎症小体可能有助于CNV的发生和发展。为了了解NLRP3炎性体在CNV中的作用，我们使用Ccr2RFPCx3cr1GFP双报告小鼠来表征激光诱导的布鲁赫膜破裂后Ccr2RFP阳性单核细胞和Cx3cr1GFP阳性小胶质细胞向CNV病变的迁移。MCC950用作NLRP3抑制剂。免疫染色用于确认NLRP3炎性体在LCNV病变中的定位。使用共聚焦显微镜对LCNV体积进行成像和定量。ELISA和qRT-PCR通过监测LCNV小鼠脉络膜组织中IL-1β蛋白和mRNA的表达来确认NLRP3的激活。此外,使用NLRP3(-/-)LCNV小鼠来研究NLRP3炎性体是否有助于LCNV病变的发展。我们观察到RFP阳性单核细胞衍生的巨噬细胞和GFP阳性小胶质细胞衍生的巨噬细胞，除了其他细胞类型，在激光损伤后第7天定位在LCNV病变中。此外,NLRP3炎性体与LCNV病变相关。抑制NLRP3炎性体，使用MCC950，导致Ccr2RFP阳性巨噬细胞增加，Cx3cr1GFP阳性小胶质细胞，和其他细胞导致总病变大小增加。NLRP3(-/-)LCNV小鼠,显示与年龄匹配的对照组相比，病变大小显着增加。抑制NLRP3，导致脉络膜组织中IL-1βmRNA和蛋白表达降低，提示病变大小增加可能与IL-1β没有直接关系。
