越橘提取物对RPE细胞氧化损伤和血管生成的抑制作用  

Inhibitory effects of bilberry extract on oxidative damage and angiogenesis in RPE cells

作　　者：刘晓雯[1] 曹琛[1] 苏映雪[1] 王延东[1] LIU Xiaowen;CAO Chen;SU Yingxue;WANG Yandong(State Key Laboratory of Ophthalmology,Zhongshan Ophthalmic Center,Sun Yat-sen University,Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science,Guangzhou 510060,China)

机构地区：[1]中山大学中山眼科中心,眼病防治全国重点实验室,广东省眼科视觉科学重点实验室,广州510060

出　　处：《眼科学报》2024年第2期84-90,共7页Eye Science

基　　金：广东省自然科学基金面上项目(2023A1515012521);广东省中医药局中医药科研项目(20231079);广州市科技重点研发计划项目(202103000050)。

摘　　要：目的:探讨越橘提取物对视网膜色素上皮(retinal pigment epithelial,RPE)细胞氧化损伤和血管生成的抑制作用。方法:①将人RPE细胞(ARPE-19)分为对照组、模型组、越橘提取物组,模型组给予0.5 mmol/L3-吗啉代亚胺(3-morpholinosydnonimine,SIN-1)处理24 h,越橘提取物组给予10ng/mL越橘提取物处理1h后,再给予0.5 mmol/L SIN-1处理24 h。细胞活性检测法(Cell Counting Kit-8,CCK-8)测定各组细胞活性,流式细胞术检测细胞活性氧(reactive oxygen species,ROS)水平。②将人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)分为对照组、模型组、越橘提取物组,模型组给予10 ng/mLVEGF处理8 h,越橘提取物组给予10 ng/mL越橘提取物处理12 h后,再给予10 ng/mLVEGF处理8 h。划痕实验和Transwell实验检测细胞迁移和侵袭,管腔形成实验检测细胞血管生成情况。结果:①越橘提取物在质量浓度≤10 ng/mL对ARPE-19细胞无明显毒性。给予SIN-1处理后,ARPE-19细胞活性明显降低,而预孵10 ng/mL越橘提取物可使细胞活性恢复接近至正常(P<0.001),后续实验均选用该浓度。同时,越橘提取物可降低SIN-1诱导的ARPE-19细胞内ROS水平(P<0.0001)。②给予VEGF处理后,HUVEC细胞迁移和侵袭能力增强,而预孵越橘提取物可使细胞迁移和侵袭能力减弱(P<0.0010)。同时,越橘提取物可抑制VEGF诱导的HUVEC细胞小管形成(P<0.0001)。结论:越橘提取物具有较强的抗氧化能力和抗血管生成活性,为其作为潜在治疗年龄相关性黄斑变性治疗药物提供了科学依据。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.

关 键 词：越橘提取物 氧化损伤 血管生成 年龄相关性黄斑变性 

分 类 号：R285.5[医药卫生—中药学]