Kavain Alleviates Choroidal Neovascularization Via Decreasing the Activity of the HIF-1α/VEGF-A/VEGFR2 Signaling Pathway and Inhibiting Inflammation

The abnormal choroidal blood vessels could lead to retinal or subretinal hemorrhage, macular edema and exudation, photoreceptor injury, fibrovascular scarring, and permanent vision loss if leftuntreated.4,5 Vascular endothelial growth factor A (VEGF) is a well-known potent growth factor in the development of ocular angiogenesis.6 Anti-VEGF therapy, including aflibercept, has been the gold standard for treating nAMD.7 However, anti-VEGF therapy faces various issues, such as drug resistance to postintravitreal injections and lack of response in certain individuals.8 The pathogenesis of nAMD is complex and remains largely unknown. Tube formation assay The tube formation assay was conducted to assess the angiogenesis of HUVECs.22 An equal density of HUVECs (5 × 104 cells per well), pretreated with drugs, was seeded in serum-free DMEM medium and cultured in a 24-well plate coated with growth-factor reduced matrigel (356230, Corning, USA) for 24 hours at 37 °C for 6 hours.23 The tube formation of HUVECs was observed and captured using a fluorescence microscope. Laser-induced CNV model After undergoing anesthesia and dilation of pupils, 8-week-old male C57BL/6J mice were subjected to laser photocoagulation treatment. The laser photocoagulation was performed using the following parameters (OcuLight GL, Iridex, Mountain View, CA): a wavelength of 532 nm, a duration of 0.1 s, power set at 120 mW, and a spot size of 50 µm. Following the laser photocoagulation procedure, the presence of a white subretinal bubble indicates the successful disruption of Bruch's membrane.