罗氟司特通过抑制 NLRP3 炎症体减轻视网膜缺血再灌注损伤后的小胶质细胞衰老和视网膜炎症性神经退行性变

IF 5 2区 医学 Q1 OPHTHALMOLOGY
Chunlian Ou, Yiwei Lin, Jing Wen, Hongyang Zhang, Ying Xu, Naiyuan Zhang, Qiong Liu, Yingzi Wu, Jing Xu, Jing Wu
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引用次数: 0

摘要

目的视网膜缺血再灌注(RIR)损伤与多种视网膜疾病有关,会导致视网膜神经节细胞(RGC)变性。小胶质细胞的衰老会加剧炎症,导致神经变性。本研究旨在探讨罗氟司特(Roflumilast,Roflu)在改善RIR损伤后小胶质细胞衰老和神经炎症方面的潜在治疗作用:方法:C57BL/6J小鼠接受RIR手术,腹腔注射罗氟司特治疗。对 BV2 小神经胶质细胞进行氧-葡萄糖剥夺和再氧合(OGD/R)以模拟体外缺血条件。SA-β-gal 染色用于检测细胞衰老。定量 PCR 和 ELISA 用于检测衰老相关分泌表型(SASP)因子的水平。对视网膜切片进行苏木精和伊红(H&E)染色,以评估视网膜的形态和厚度。使用免疫荧光(IF)对视网膜全切片中存活的 RGC 进行标记和量化。此外,还使用了 Western 印迹和 IF 染色来量化与细胞周期和 NLRP3 炎症小体相关的蛋白质:结果:罗福治疗减少了小胶质细胞的衰老、ROS的产生以及OGD/R-暴露的BV2细胞中促炎细胞因子的分泌。它还能恢复细胞增殖能力,逆转 OGD/R 诱导的细胞周期停滞。在 RIR 小鼠模型中,罗弗卢减轻了视网膜衰老,保护了视网膜厚度,并防止了 RGCs 的死亡。从机理上讲,罗非鲁抑制了小胶质细胞中NLRP3炎性体的激活,并抑制了DNA损伤信号通路:结论:在 RIR 损伤中,Roflu 可通过抑制 NLRP3 炎性体减轻小胶质细胞的衰老和炎症反应,从而发挥神经保护作用。这些研究结果表明,Roflu 可通过靶向小胶质细胞衰老,作为一种治疗缺血性损伤相关视网膜疾病的有效策略。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Roflumilast Attenuates Microglial Senescence and Retinal Inflammatory Neurodegeneration Post Retinal Ischemia Reperfusion Injury Through Inhibiting NLRP3 Inflammasome.

Purpose: Retinal ischemia-reperfusion (RIR) injury is implicated in various retinal diseases, leading to retinal ganglion cells (RGCs) degeneration. Microglial senescence exacerbates inflammation, contributing to neurodegeneration. This study aimed to investigate the potential therapeutic role of Roflumilast (Roflu) in ameliorating microglial senescence and neuroinflammation following RIR injury.

Methods: C57BL/6J mice underwent RIR surgery, and Roflu treatment was administered intraperitoneally. BV2 microglial cells were subjected to oxygen-glucose deprivation and reoxygenation (OGD/R) to simulate ischemic conditions in vitro. SA-β-gal staining was used to detect cellular senescence. Quantitative PCR and ELISA were used to examine the levels of senescence-associated secretory phenotype (SASP) factors. Hematoxylin and eosin (H&E) staining was performed on retinal sections to assess retinal morphology and thickness. Surviving RGCs were labeled and quantified in retinal whole-mounts using immunofluorescence (IF). Furthermore, Western blot and IF staining were used to quantify the proteins associated with the cell cycle and NLRP3 inflammasomes.

Results: Roflu treatment reduced microglial senescence, ROS production, and secretion of pro-inflammatory cytokines in OGD/R-exposed BV2 cells. It also restored cell proliferation capacity and reversed OGD/R-induced cell cycle arrest. In vivo, Roflu alleviated retinal senescence, preserved retinal thickness, and protected against RGCs death in the RIR mouse model. Mechanistically, Roflu inhibited the NLRP3 inflammasome activation and suppressed DNA damage signaling pathway in microglia.

Conclusions: Roflu exerts neuroprotective effects by mitigating microglial senescence and inflammation via inhibition of the NLRP3 inflammasome in RIR injury. These findings suggest that Roflu may serve as a promising therapeutic strategy for retinal diseases associated with ischemic injury by targeting microglial senescence.

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来源期刊
CiteScore
6.90
自引率
4.50%
发文量
339
审稿时长
1 months
期刊介绍: Investigative Ophthalmology & Visual Science (IOVS), published as ready online, is a peer-reviewed academic journal of the Association for Research in Vision and Ophthalmology (ARVO). IOVS features original research, mostly pertaining to clinical and laboratory ophthalmology and vision research in general.
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