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 0.5mmol/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 10ng/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: 1. Bilberry extract had no obvious toxicity to ARPE-19 cells(≤10 ng/mL). AfterSIN-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, whil epretreatment 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.

目的：观察弱内向整流相关及花生四烯酸激活的弱内向整流相关双孔钾离子通道（tandem of pore domains in a weak inward rectifying related-tandem of pore domains in a weak inward rectifying related arachidonic acid activated two pore-domain potassium channels，TREK-TRAAK K2P）激活对氧化损伤的人视网膜色素上皮细胞（retinal pigment epithelial, RPE）吞噬功能的影响。方法：免疫荧光法检测TREK-1、TREK-2及TRAAK通道蛋白在人RPE细胞的表达，以不同时间和浓度梯度的叔丁基过氧化氢（tert-butyl hydroperoxide, t-BHP）诱导人RPE细胞氧化损伤。分对照组、t-BHP组、利鲁唑加t-BHP组，利鲁唑组，向各组加入2×107/mL的荧光微球或FITC标记猪感光细胞外节膜盘孵育6 h，固定染色后荧光显微镜拍照，Image-Pro Plus 6.0软件分析计算RPE细胞吞噬率和吞噬指数。结果：TREK-1、TREK-2、TREEK通道蛋白亚型在人RPE细胞质中均高表达。人RPE细胞在t-BHP干预后存活率呈浓度和时间依赖性，200 μmol/L t-BHP干预6 h与对照组存活率比较差异无统计学意义（P＞0.05），400μmol/L t-BHP干预6 h致半数死亡。特异性吞噬指数：t-BHP组低于其他各组（P＜0.001），而利鲁唑组高于对照组，差异无统计学意义（P＞0.05）。特异性吞噬率：t-BHP组低于其他各组，差异无统计学意义（P＞0.05）。非特异性吞噬指数：t-BHP组低于其他各组（P＜0.001）；非特异性吞噬率：t-BHP组低于利鲁唑加t-BHP组，差异无统计学意义（P＞0.05），两两比较，仅利鲁唑组高于t-BHP组（P＜0.05）。结论：激活TREK-TRAAK K2P通道可保护氧化损伤人RPE细胞的吞噬功能。  

Abstract Objective: To observe the effect of TREK-TRAAK K2P activation on the phagocytic function of oxidative damaged human retinal pigment epithelial cells (RPE). Methods: Immunofluorescence staining was used to detect the expression of TREK-1, TREK-2 and TRAAK channel proteins in human RPE cells, and the RPE cell oxidative damage model was induced by tert-butyl hydroperoxide (t-BHP) at different time and concentration gradients. The study was divided into control group, t-BHP model group, riluzole plus t-BHP group, and riluzole group. 2×107/mL fluorescent microspheres or FITC-labeled porcine photoreceptor outer segment membrane discs were added to each group for 6h incubation, fluorescence photos were obtained after fixation and staining, and the phagocytosis rate and phagocytosis index of RPE cells was analyzed and calculated by Image-Pro Plus 6.0 software. Results: TREK-1, TREK-2, and TREEK channel subtype proteins were all highly expressed in the human RPE cytoplasm. The survival rate of human RPE cells after t-BHP intervention was concentration- and time-dependent. There was no statistical difference in the survival rate between 200 μmol/L t-BHP intervention for 6 hours and the control group (P>0.05), and 400 μmol/L t-BHP intervention for 6 hours induced half death. Specific phagocytic index: The t-BHP group was lower than other groups (P<0.001). Specific phagocytosis rate: the t-BHP group was lower than other groups, with no statistical difference (P>0.05). Nonspecific phagocytic index: the t-BHP group was lower than other groups (P<0.001); nonspecific phagocytic rate: the t-BHP model group was lower than the riluzole plus t-BHP group, with no statistical difference (P>0.05), and only the riluzole group was higher than the t-BHP model group (P＜0.05) in pairwise comparison. Conclusions: Activation of TREK-TRAAK K2P can protect the phagocytic function of human RPE cells damaged by oxidative stress.