Inhibitory effects of bilberry extract on oxidative damage and angiogenesis in RPE cells
Objective:To investigate the inhibitory effects of bilberry extract on oxidative damage and angiogenesis in retinal pigment epithelial(RPE)cells.Methods:Human RPE cells(ARPE-19)were divided into control,model and bilberry extract groups.The model group was treated with 0.5 mmol/L SIN-1 for 24 h,and the bilberry extract group was treated with 10ng/bilberry extract for 1 hour,followed by 0.5 mmol/L SIN-1 for 24 h.Cell viability was measured by CCK-8.Level of reactive oxygen species(ROS)was determined using flow cytometry.Human umbilical vein endothelial cells(HUVEC)were divided into three groups,control group,model group and bilberry extract group.The model group was treated with 10 ng/mL VEGF for 8 h,and the bilberry extract group was treated with 10 ng/mL bilberry extract for 12 h,followed by 10ng/mLVEGF for 8 h.Cell migration and invasion were measured by wound healing assay and Transwell assay.Cell angiogenesis was determined by tube formation assay.Results:Bilberry extract had no obvious toxicity to ARPE-19 cells(≤10 ng/mL).After SIN-1 treatment,the the viability of ARPE-19 cells was significantly reduced,while incubation with 10 ng/mL bilberry extract could restore cell activity to the normal levels(P<0.001).This concentration was selected for subsequent experiments.Additionally,bilberry extract reduced the level of ROS in SIN-1-induced ARPE-19 cells(P<0.0001).VEGF treatment significantly enhanced the migration and invasion of HUVEC,while pretreatment with bilberry extract attenuated these effects(P<0.0010).Meanwhile,bilberry extract could significantly inhibit VEGF-induced tube formation in HUVEC(P<0.0001).Conclusion:Bilbery extract possess strongantioxidant and anti-angiogenetic activities,suggesting its potential as treatment agent for AMD.
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