Department of Ophthalmology, People's Hospital of Guangxi Zhuang Autonomous Region, Nanning, 530021, Guangxi, People's Republic of China.
Department of Ophthalmology, Renmin Hospital of Wuhan University, Wuhan, 430060, Hubei, People's Republic of China.
J Mol Neurosci. 2018 Feb;64(2):200-210. doi: 10.1007/s12031-017-1020-z. Epub 2017 Dec 26.
To explore the hypothesis that CD200Fc, a CD200R1 agonist with anti-inflammatory properties, will inhibit retinal glial cells hyperactivation and retinal ganglion cells (RGCs) apoptosis after optic nerve injury. CD200Fc was immediately administered after optic nerve crush (ONC) once by intravitreal injection. Rats were euthanized at 5 days after ONC. The density of RGCs was counted by immunostaining of retina flat mounts for Brn3a. TUNEL assay, immunoblotting analysis of ionized calcium-binding adapter molecule 1(iba1) (microglia marker) and glial fibrillary acidic protein (GFAP) (astrocytes and Müller cells marker), RT-PCR analysis of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS), monocyte chemotactic protein 1 (MCP-1), tumor necrosis factor-α (TNF-α), interleukin (IL)-8 and IL-10, ELISA measure protein levels of inflammatory cytokines and western blot analysis of CD200 and CD200R1 were evaluated. CD200Fc treatment suppressed ONC-induced RGCs loss through inhibition of RGCs apoptosis. Additionally, expression of glial cells activation markers GFAP and iba1 and production of pro-inflammatory cytokines (COX-2, iNOS, MCP-1, TNF-α, IL-8) were decreased in CD200Fc treated animals after ONC. Meanwhile, anti-inflammatory cytokine IL-10 was increased by CD200Fc treatment in ONC-induced rat retina. Finally, we found that CD200Fc significantly inhibited ONC-induced increased in expression of CD200 and raised the already high basal CD200R1 expression in the rat retina after ONC. Our results demonstrated that the anti-inflammatory effects of CD200Fc in ONC rats model through inhibited the activation of retinal glial cells via the interaction between CD200 and CD200R1, and the neuroprotective effects of CD200Fc on RGCs thought inhibited its apoptosis.
CD200Fc(一种具有抗炎特性的 CD200R1 激动剂)可抑制视神经损伤后视网膜神经胶质细胞的过度激活和视网膜神经节细胞(RGCs)的凋亡。视神经挤压(ONC)后,通过玻璃体内注射立即给予 CD200Fc。ONC 后 5 天处死大鼠。通过视网膜铺片 Brn3a 的免疫染色计数 RGCs 的密度。TUNEL 检测、离子钙结合接头分子 1(iba1,小胶质细胞标志物)和胶质纤维酸性蛋白(GFAP,星形胶质细胞和 Müller 细胞标志物)的免疫印迹分析、环氧化酶-2(COX-2)、诱导型一氧化氮合酶(iNOS)、单核细胞趋化蛋白 1(MCP-1)、肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-8 和 IL-10 的 RT-PCR 分析、炎症细胞因子蛋白水平的 ELISA 测定以及 CD200 和 CD200R1 的 Western blot 分析。CD200Fc 治疗通过抑制 RGCs 凋亡抑制了 ONC 诱导的 RGCs 丢失。此外,在 ONC 后,CD200Fc 处理的动物中,GFAP 和 iba1 等神经胶质细胞激活标志物的表达以及促炎细胞因子(COX-2、iNOS、MCP-1、TNF-α、IL-8)的产生减少。同时,在 ONC 诱导的大鼠视网膜中,CD200Fc 治疗增加了抗炎细胞因子 IL-10 的表达。最后,我们发现 CD200Fc 显著抑制了 ONC 诱导的 CD200 表达增加,并在 ONC 后提高了大鼠视网膜中已经很高的基础 CD200R1 表达。我们的研究结果表明,CD200Fc 通过抑制 CD200 与 CD200R1 的相互作用抑制视网膜神经胶质细胞的激活,在 ONC 大鼠模型中发挥抗炎作用,通过抑制其凋亡对 RGCs 发挥神经保护作用。