Ang-1对高糖环境中猴脉络膜-视网膜内皮细胞单层通透性的影响  

Effects of angiopoietin-1 on rhesus macaque choroids-retinal endothelial cells monolayer permeability in high glucose environment

作　　者：萧鸿[1] 左中夫[2] 冯闯[3] 刘学政[4] 

机构地区：[1]辽宁医学院附属第一医院教务科,辽宁锦州121001 [2]辽宁医学院病理生理教研室,辽宁锦州121001 [3]辽宁医学院研究生学院,辽宁锦州121001 [4]辽宁医学院人体解剖学教研室,辽宁锦州121001

出　　处：《第四军医大学学报》2009年第3期195-198,共4页Journal of the Fourth Military Medical University

基　　金：辽宁省优秀青年科研人才培养资金资助项目(3050007);辽宁省科技计划资助项目(2003225007)

摘　　要：目的:利用猴脉络膜-视网膜内皮细胞建立内皮细胞(EC)单层模型,观察高浓度葡萄糖及血管生成素-1(Ang-1)对该模型通透性的影响.方法:待EC于孔径0.8μm的聚碳酸酯微孔滤膜上形成致密单层后分3组进行培养:对照组(DMEM培养基含25 mmol/L葡萄糖)、高糖组(DMEM培养基含30 mmol/L葡萄糖)、Ang-高糖组(DMEM培养基含30mmol/L葡萄糖+0.25 mg/L Ang-1).于1,3,5,7 d 4个时间点,以含白蛋白1 g/L的D-Hanks液灌注滤膜,检测蛋白质渗透压反射系数(δ)及滤过系数(Kf);并于5 d和7 d利用琼脂糖凝胶电泳检测DNA ladder.结果:3,5,7 d时,高糖组EC单层的通透性高于对照组(P<0.01);Ang-高糖组与对照组之间EC单层的通透性无差异(P>0.05).高糖组EC的琼脂糖凝胶电泳在5 d及7 d均出现DNA ladder,对照组及Ang-高糖组未见DNA ladder.结论:高糖能够促进细胞凋亡,增加EC单层通透性;Ang-1能抑制EC的凋亡,对抗高糖所致的EC单层通透性增高.AIM： To explore the effects of high concentration glucose and angiopoietin-1 （ Ang-1 ） on monolayer permeability of rhesus macaque choroids-retinal endothelial cells （ECs）. METHODS ： Confluent ECs monolayer was developed on polyearbonate mieropore filter. The cells were divided into 3 groups： control group （ DMEM containing 25 mmol/L glucose） , high glucose group （ DMEM containing 30 mmol/L glucose） and Ang-1 ＋ high glucose group （ DMEM containing both 30 mmol/L glucose and 0.25 mg/L Ang-1 ）. The ECs were perfused with plain D- Hank＇s balanced salt solution containing 1 g/L albumin to detect the fluid filtration coefficient （Kf） and osmotic inflation coefficient （8） to the albumin at the 1st, 3rd, 5th and 7th day, respectively. Additionally, DNA agarose gel eleetrophoresis was performed at the 5th and 7th day, respectively. RESULTS ： The monolayer permeability of ECs in the high glucose group was higher than that in control group at the 3rd, 5th and 7th day （P 〈 0.01 ）. But no significant difference was observed in the permeability between control group and Ang-glucose group （ P 〉 0.05 ）. DNA ladder was only presented in high glucose group at the 5th and 7th day. CONCLUSION： High concentration glucose increases the EC monolayer permeability by accelerating the speed of apoptosis of ECs. Ang-1 may prevent ECs from apoptosis and oppose the effects of high glucose on EC monolayer permeability.

关 键 词：脉络膜 视网膜内皮细胞 血管生成素-1 高糖 通透性 猴 

分 类 号：R774.1[医药卫生—眼科]