本研究的目的是探讨 P2X7 受体拮抗剂 A740003 对慢性高眼压 (COH) 实验性青光眼小鼠模型视网膜神经节细胞 (RGC) 的神经保护作用。利用生物信息学分析青光眼相关基因。采用Western blot、实时荧光定量PCR和免疫荧光染色技术探讨A740003对COH视网膜RGC的神经保护作用机制。生物信息分析显示，氧化应激、神经炎症、细胞凋亡与青光眼的发病密切相关。在 COH 视网膜中，眼压升高显着增加了易位蛋白的水平，这是小胶质细胞激活的标志物，可以通过玻璃体内预注射 A740003 来逆转这种情况。 A740003 还抑制 COH 视网膜中促炎细胞因子白细胞介素 (IL) 1β 和肿瘤坏死因子 α mRNA 水平的增加。此外，虽然COH视网膜中抗炎细胞因子IL-4和IL-10的mRNA水平保持不变，但施用A740003可以增加它们的水平。 COH 视网膜中 Bax 和 cleaved caspase-3 的 mRNA 和蛋白水平增加，A740003 可以部分逆转这种情况，而注射或不注射 A740003 的 COH 视网膜中 Bcl-2 水平保持不变。此外，A740003 部分减弱了 COH 小鼠中 Brn-3a 阳性 RGC 数量的减少。 A740003 可以通过抑制小胶质细胞激活、减弱视网膜炎症反应、减少 RGC 凋亡以及增强 COH 实验性青光眼中 RGC 的存活来对 RGC 提供神经保护作用。版权所有 © 2024 Wolters Kluwer Health, Inc. 保留所有权利。

The aim of this study was to explore the neuroprotective effects of the P2X7 receptor antagonist A740003 on retinal ganglion cells (RGCs) in chronic intraocular hypertension (COH) experimental glaucoma mouse model. Bioinformatics was used to analyze the glaucoma-related genes. Western blot, real-time fluorescence quantitative PCR, and immunofluorescence staining techniques were employed to explore the mechanisms underlying the neuroprotective effects of A740003 on RGCs in COH retinas. Bioinformatic analysis revealed that oxidative stress, neuroinflammation, and cell apoptosis were highly related to the pathogenesis of glaucoma. In COH retinas, intraocular pressure elevation significantly increased the levels of translocator protein, a marker of microglial activation, which could be reversed by intravitreal preinjection of A740003. A740003 also suppressed the increased mRNA levels of proinflammatory cytokines interleukin (IL) 1β and tumor necrosis factor α in COH retinas. In addition, although the mRNA levels of anti-inflammatory cytokine IL-4 and IL-10 were kept unchanged in COH retinas, administration of A740003 could increase their levels. The mRNA and protein levels of Bax and cleaved caspase-3 were increased in COH retinas, which could be partially reversed by A740003, while the levels of Bcl-2 kept unchanged in COH retinas with or without the injections of A740003. Furthermore, A740003 partially attenuated the reduction in the numbers of Brn-3a-positive RGCs in COH mice. A740003 could provide neuroprotective roles on RGCs by inhibiting the microglia activation, attenuating the retinal inflammatory response, reducing the apoptosis of RGCs, and enhancing the survival of RGCs in COH experimental glaucoma.Copyright © 2024 Wolters Kluwer Health, Inc. All rights reserved.