Ang-1对高糖环境中RF/6A单层的保护作用  

The protection of angiopoietin-1 on monolayer of endothelial cells exposed to high glucose

作　　者：刘学政[1] 左中夫[1] 冯闯[1] 王轶晶[2] 

机构地区：[1]辽宁医学院人体解剖学教研室,锦州121001 [2]辽宁医学院生物技术研究所

出　　处：《眼科研究》2009年第5期380-384,共5页Chinese Ophthalmic Research

摘　　要：目的探讨高浓度葡萄糖及血管生成素-1(Ang-1)对内皮细胞(EC)单层通透性的影响。方法将猴脉络膜-视网膜内皮细胞(RF/6A)接种于0.8μm聚碳酸酯微孔滤膜,形成致密单层后分3个组培养。正常对照组:含葡萄糖25mmol/L;高糖组:含葡萄糖40mmol/L;Ang-高糖组:含葡萄糖40mmol/L+Ang-1(250ng/mL)。于1、3、5、7d,4个时间点建立EC单层通透性模型,检测蛋白质渗透压反射系数(δ)及滤过系数(Kf),以监测EC单层通透性的变化;利用Westernblot(NBT/BCIP显色法)检测5d时survivin的表达。结果3、5、7d时,高糖组EC单层通透性增高(P<0.01),Ang-高糖组在5d及7d时,EC单层通透性升高(P<0.01),但仍低于高糖组;Westernblot结果显示,Ang-高糖组survivin表达明显高于其他各组(P<0.01)。结论高糖能够增加EC单层通透性,Ang-1能对抗高糖的这种生物学作用。可能是通过上调凋亡抑制基因survivin的表达,抑制RF/6A的凋亡而发挥生物学作用的。Objective This study aimed to explore the effects of angiopoietin-1 （ Aug-1 ） and high glucose on rhesus macaque choroids-retinal endothelial cells （RF/6A）. Methods RF/6A cells cultured on polycarbonate mieropore filter membrane were treated with DMEM containing 25,40 mmol/L glucose or 40 mmol/L glucose ＋ 0.25 ng/L Ang-1 respectively （control group,high glucose group and Ang-glueose group）. After the formation of monolayer of RF/6A cells, the fluid filtration coefficient（Kf） and osmotic inflation coefficient （3） to protein were detected by perfusion of plain Hank＇ s balanced saline solution containing 1 g/L albumin at the 1st, 3rd, 5th and 7th day respectively. Western blot was performed to determine the expression of survivin in the dense monolayer of RF/6A at the 5th day. Results The osmotic inflation coefficient（δ value） of dense monolayer of RF/6A was significantly different in various groups, and the δ value was significantly decreased in the high glucose group compared with control group and Aug-glucose group at the 3rd,5th and 7th day（ P 〈 0. 01 ,P 〈 0.05）. The 8 value was obviously reduced in Ang-glucose group compared with normal control group at the 5th and 7th day（ P 〈 0.05 ）. The Kf value of dense monolayer of RF/6A was considerably different in various groups. In high glucose group compared with normal group and Ang-glucose group,the Kf value was higher （P 〈 0. 01 , P 〈 0. 05 ） . Western blot showed significant difference among the four groups and that of Aug-glucose group was obviously elevated in comparison with normal control group（ P 〈 0. 01 ）. Conclusion High glucose increase the permeability of monolayer epithelial cells, and Aug-1 plays the beneficial effect in limiting this damage by up-regulating the expression of surviving and therefore preventing RF/6A from apoptosis.

关 键 词：猴脉络膜-视网膜内皮细胞 血管生成素-1 高糖 内皮细胞单层 通透性 SURVIVIN 

分 类 号：R587.1[医药卫生—内分泌] R774.1[医药卫生—内科学]