Institute of Neuroscience and Third Affiliated Hospital, Zhengzhou University.
Academy of Medical Sciences, Zhengzhou University, Zhengzhou.
Neuroreport. 2024 Sep 4;35(13):822-831. doi: 10.1097/WNR.0000000000002071. Epub 2024 Jul 8.
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.
本研究旨在探讨 P2X7 受体拮抗剂 A740003 对慢性眼内高压（COH）实验性青光眼小鼠模型中视网膜神经节细胞（RGC）的神经保护作用。采用生物信息学方法分析与青光眼相关的基因。运用 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 神经保护作用的机制。生物信息学分析表明，氧化应激、神经炎症和细胞凋亡与青光眼的发病机制密切相关。在 COH 视网膜中，眼压升高显著增加了转位蛋白的水平，这是小胶质细胞激活的标志物，而 A740003 可以逆转这种情况。A740003 还抑制了 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的 mRNA 水平升高。此外，尽管 COH 视网膜中抗炎细胞因子 IL-4 和 IL-10 的 mRNA 水平保持不变，但给予 A740003 可以增加它们的水平。Bax 和 cleaved caspase-3 的 mRNA 和蛋白水平在 COH 视网膜中升高，A740003 可部分逆转这种情况，而 Bcl-2 的水平在 COH 视网膜中保持不变，无论是否注射 A740003。此外，A740003 部分减轻了 COH 小鼠中 Brn-3a 阳性 RGC 数量的减少。A740003 通过抑制小胶质细胞激活、减轻视网膜炎症反应、减少 RGC 凋亡以及增强 COH 实验性青光眼的 RGC 存活，对 RGC 发挥神经保护作用。
