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：探讨血管生成拟态(vasculogenic mimicry，VM)与翼状胬肉初发型及复发型的相关性。方法：采用血小板-内皮细胞黏附分子/过碘酸雪夫(platelet endothelial cell adhesion molecule-1/periodic acid-schiff，CD31/PAS)免疫组织化学双重染色法检测139例翼状胬肉组织(初发型105例；复发型34例)和10例正常结膜中VM的表达，分析VM与初发型及复发型翼状胬肉的相关性及其与患者性别、年龄等因素的关系。原代培养人翼状胬肉成纤维细胞(human pterygium fibroblasts，HPFs)，免疫细胞化学染色法鉴定，利用三维培养及PAS染色观察初发型和复发型HPFs构成VM管腔个数的差异。结果：10例正常结膜均未见VM结构，初发型翼状胬肉VM阳性率43.81%，复发型翼状胬肉VM阳性率82.35%，差异具有统计学意义(P<0.001)。相关性分析显示VM与复发型翼状胬肉呈显著正相关(r=0.332)。不同性别、年龄及病程的翼状胬肉患者VM的表达差异均无统计学意义(均P>0.05)。原代培养的HPFs Vimentin表达阳性，符合成纤维细胞特性。细胞三维培养及PAS染色结果提示HPFs具有构建体外VM模型的能力，且复发型HPFs构成的VM管腔数明显高于初发型，差异具有统计学意义(P<0.01)。结论：翼状胬肉组织中存在VM结构，可作为其血供途径之一。VM与翼状胬肉的复发具有密切关系。

Objective: The purpose of this study was to investigate the correlation of vasculogenic mimicry in the primary and recurrent pterygium. Methods: Platelet endothelial cell adhesion molecule-1/periodic acid-schiff (CD31/PAS)immunohistochemical double staining method was adopted to detect the expression of VM in 139 cases of pterygium (105 cases of primary pterygium and 34 cases of recurrent pterygium)and 10 cases of normal conjunctival tissues. The correlation between VM and primary pterygium, recurrent pterygium and the factors such as gender and age of patients were analyzed. Human pterygium fibroblasts (HPFs) were primary cultured and identified by immunocytochemical staining. The differences in the number of VM channels between primary HPFs and recurrent HPFs were observed by three-dimensional culture and PAS staining. Results: There was no VM structure in 10 normal conjunctiva and the positive rate of VM was 43.81% in primary pterygium and 82.35% in recurrent pterygium with a significantly difference (P<0.001). Correlation analysis showed a significant positive correlation between VM and recurrent pterygium (r=0.332). There was no significant difference in the expression of VM in pterygium patients with different sex, age and course (all P>0.05). Vimentin was positive in the primary cultured cells, which was consistent with the characteristics of fibroblasts. The results of three-dimensional culture and PAS staining indicated that HPFs had the ability to construct VM model in vitro, and the number of VM channels constituted by recurrent HPFs was significantly higher than that by primary HPFs, the difference was statistically significant (P<0.01). Conclusion: VM exists in pterygium tissues, and it can be used as one of the blood supply routes, which is closely related to the recurrence of pterygium.