Chen Wanjing, Iizuka Yoko, Mabuchi Fumihiko, Kashiwagi Kenji
Department of Ophthalmology, School of Medicine, University of Yamanashi, Yamanashi 409-3898, Japan.
J Clin Med. 2025 Jul 29;14(15):5344. doi: 10.3390/jcm14155344.
: To investigate the neuroprotective effects of a Rho-associated kinase (ROCK) inhibitor on retinal ganglion cells (RGCs) in vitro and in vivo. : For in vivo studies, a unilateral optic nerve crush mouse model was established. Then, 100 mM Y-27632 (a ROCK inhibitor) or saline was applied to the experimental eyes once a day for 14 days. The effects of the ROCK inhibitor were evaluated by counting the surviving RGCs in the enucleated flat retina tissues and measuring the inner retinal thickness using optical coherence tomography (OCT), the amplitude of the electroretinogram (ERG), and the change in intraocular pressure (IOP). For the in vitro study, RGCs were isolated from five-day-old mice using a modified immunopanning method with magnetic beads. The isolated RGCs were incubated for 72 h with various concentrations of Y-27632, after which TUNEL assays were performed to determine the number of surviving RGCs. : Y-27632 has neuroprotective effects, as it significantly increased the number of surviving RGCs by approximately 6.3%. OCT and ERG data also revealed that Y-27632 induced neuroprotective effects in vivo; furthermore, Y-27632 reduced IOP by approximately 18.3%. The in vitro study revealed the dose-dependent neuroprotective effects of Y-27632, with the highest dose of Y-27632 (1000 nM) increasing the RGC survival rate after 72 h of incubation compared with that of the control. : The ROCK inhibitor Y-27632 may exert some neuroprotective effects on RGCs when it is used as an eye drop through an IOP-independent mechanism.
研究Rho相关激酶（ROCK）抑制剂对视网膜神经节细胞（RGCs）的体内外神经保护作用。
体内研究采用单侧视神经挤压小鼠模型。然后，每天一次向实验眼应用100 mM Y-27632（一种ROCK抑制剂）或生理盐水，持续14天。通过计数摘除眼球的扁平视网膜组织中存活的RGCs数量、使用光学相干断层扫描（OCT）测量视网膜内层厚度、视网膜电图（ERG）的振幅以及眼内压（IOP）的变化来评估ROCK抑制剂的作用。体外研究中，采用改良的磁珠免疫淘选法从5日龄小鼠分离RGCs。将分离的RGCs与不同浓度的Y-27632孵育72小时，之后进行TUNEL检测以确定存活的RGCs数量。
Y-27632具有神经保护作用，因为它使存活的RGCs数量显著增加了约6.3%。OCT和ERG数据还显示Y-27632在体内具有神经保护作用；此外，Y-27632使IOP降低了约18.3%。体外研究揭示了Y-27632的剂量依赖性神经保护作用，与对照组相比，最高剂量的Y-27632（1000 nM）在孵育72小时后提高了RGCs的存活率。
ROCK抑制剂Y-27632作为滴眼液使用时，可能通过与眼压无关的机制对RGCs发挥一定的神经保护作用。
