Zhang Hui-Ming, Li Xiao-Hua, Chen Mingjiazi, Luo Jing
Department of Dermatology, the Second Xiangya Hospital, Central South University, Changsha 410011, Hunan Province, China.
Hunan Key Laboratory of Medical Epigenomics, Changsha 410011, Hunan Province, China.
Int J Ophthalmol. 2020 Jun 18;13(6):886-892. doi: 10.18240/ijo.2020.06.05. eCollection 2020.
To determine the effects of intravitreal resveratrol (RSV) on murine laser-induced choroidal neovascularization (CNV).
The toxicity of RSV to choroidal endothelial cell (CEC) was measured using thiazolyl blue tetrazolium bromide (MTT) assay. Effects of RSV on choroidal endothelial cell (CEC) migration were evaluated with a modified Boyden chamber assay, while tube formation was evaluated in a 2-D gel assay. CNV was induced by laser photocoagulation in mice. The effects of intravitreal injection of RSV on CNV development were evaluated by fluorescein angiography (FA), confocal analysis of isolectin B4 labeled choroidal flat mounts, and histologic examination of CNV membranes. Immunostaining was used to analyze the expression and phosphorylation of vascular endothelial growth factor receptor 2 (VEGFR2).
No significant cell toxicity was observed in CEC if the concentration of RSV was less than 200 µmol/L (>0.05). RSV inhibited vascular endothelial growth factor (VEGF)-induced CEC migration (<0.05) and tube formation (<0.05) . Furthermore, intravitreal injection of RSV significantly inhibited laser induced CNV formation in mice. The FA leakage, CNV volume and CNV area analysis revealed that there were 41%, 45%, and 58% reduction in RSV-treated eyes (1.691±0.1032, 178 163±78 623 µm and 6508±619.0 µm, respectively) compared with those in control (2.724±0.08447, 379 676±98 382 µm and 16 576±2646 µm, respectively; <0.05). Phospho-VEGFR2 expression was much weaker in the sections of CNV lesions in RSV injected mice compared with that in control (<0.05).
Intravitreal injection of RSV exerts an inhibitory effect on CNV, which may through suppressing endothelial cell migration, tube formation and VEGFR2 phosphorylation.
确定玻璃体内注射白藜芦醇（RSV）对小鼠激光诱导脉络膜新生血管形成（CNV）的影响。
采用噻唑蓝四唑溴盐（MTT）法检测RSV对脉络膜内皮细胞（CEC）的毒性。用改良的Boyden小室法评估RSV对脉络膜内皮细胞（CEC）迁移的影响，同时在二维凝胶试验中评估其对血管生成的影响。通过激光光凝诱导小鼠形成CNV。通过荧光素血管造影（FA）、异凝集素B4标记的脉络膜扁平标本的共聚焦分析以及CNV膜的组织学检查，评估玻璃体内注射RSV对CNV发展的影响。采用免疫染色分析血管内皮生长因子受体2（VEGFR2）的表达和磷酸化情况。
当RSV浓度低于200µmol/L时，未观察到对CEC有明显的细胞毒性（>0.05）。RSV抑制血管内皮生长因子（VEGF）诱导的CEC迁移（<0.05）和血管生成（<0.05）。此外，玻璃体内注射RSV可显著抑制小鼠激光诱导的CNV形成。FA渗漏、CNV体积和CNV面积分析显示，与对照组相比，RSV治疗组的眼睛分别减少了41%、45%和58%（分别为1.691±0.1032、178 163±78 623µm和6508±619.0µm）（对照组分别为2.724±0.08447、379 676±98 382µm和16 576±2646µm；<0.05）。与对照组相比，RSV注射小鼠CNV病变切片中磷酸化VEGFR2的表达明显减弱（<0.05）。
玻璃体内注射RSV对CNV具有抑制作用，其机制可能是通过抑制内皮细胞迁移、血管生成和VEGFR2磷酸化来实现。
