Intravitreal injection of resveratrol inhibits laser-induced murine choroidal neovascularization

Hui-Ming Zhang; Xiao-Hua Li; Mingjiazi Chen; Jing Luo

doi:10.18240/ijo.2020.06.05

Intravitreal injection of resveratrol inhibits laser-induced murine choroidal neovascularization

Int J Ophthalmol. 2020 Jun 18;13(6):886-892. doi: 10.18240/ijo.2020.06.05. eCollection 2020.

Authors

Hui-Ming Zhang^{1

2

3}, Xiao-Hua Li⁴, Mingjiazi Chen⁵, Jing Luo⁵

Affiliations

¹ Department of Dermatology, the Second Xiangya Hospital, Central South University, Changsha 410011, Hunan Province, China.
² Hunan Key Laboratory of Medical Epigenomics, Changsha 410011, Hunan Province, China.
³ Department of Pathology and Ophthalmology, Keck School of Medicine at University of Southern California, Los Angeles, California 90033, USA.
⁴ Henan Provincial People's Hospital; Henan Eye Hospital, Henan Eye Institute, Henan Key Laboratory of Ophthalmology and Vision Science; Department of Ophthalmology, People's Hospital of Zhengzhou University; Department of Ophthalmology, People's Hospital of Henan University, Zhengzhou 450003, Henan Province, China.
⁵ Department of Ophthalmology, the Second Xiangya Hospital, Central South University, Changsha 410011, Hunan Province, China.

Abstract

Aim: To determine the effects of intravitreal resveratrol (RSV) on murine laser-induced choroidal neovascularization (CNV).

Methods: 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).

Results: No significant cell toxicity was observed in CEC if the concentration of RSV was less than 200 µmol/L (P>0.05). RSV inhibited vascular endothelial growth factor (VEGF)-induced CEC migration (P<0.05) and tube formation (P<0.05) in vitro. 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 µm³ and 6508±619.0 µm², respectively) compared with those in control (2.724±0.08447, 379 676±98 382 µm³ and 16 576±2646 µm², respectively; P<0.05). Phospho-VEGFR2 expression was much weaker in the sections of CNV lesions in RSV injected mice compared with that in control (P<0.05).

Conclusion: Intravitreal injection of RSV exerts an inhibitory effect on CNV, which may through suppressing endothelial cell migration, tube formation and VEGFR2 phosphorylation.

Keywords: choroidal neovascularization; intravitreal injection; murine; resveratrol; tube formation; vascular endothelial growth factor receptor 2.

International Journal of Ophthalmology Press.