摘要
目的探讨高浓度葡萄糖对内皮细胞(endothelial cell,EC)单层通透性模型通透性的影响以及血管生成素1(angiopoietin-1,Ang-1)对此的保护作用。方法将猴脉络膜-视网膜内皮细胞(RF/6A)接种于孔径0.8μm聚碳酸酯微孔滤膜中,形成致密单层后分三组培养。正常对照组:DMEM高糖培养基,含葡萄糖25mmol/L;;高糖组:DMEM培养基,含葡萄糖40mmol/L;;Ang-高糖组:DMEM培养基,含葡萄糖40mmol/L+Ang-1250ng/ml。于第1天、第3天、第5天、第7天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)检测第5天时survivin的表达。结果第3天、第5天及第7天时,高糖组EC单层通透性增高(P<0.01),Ang-高糖组在第5天及第7天时,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。它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。
Objective To explore the effects of angiopoietin-1 （Ang-1） and high-concentration glucose on rhesus macaque choroid-retinal endothelial cells （RF/6A） with a model for detecting EC monolayer permeability. Methods RF/6A cultured on polycarbonate micropore filter membranes were divided into 3 groups （with 25 and 40 mmol/L glucose and 40 mmol/L glucose together with 250 ng/ml Ang-1）. After the confluent EC monolayers were developed, they were used to detect the fluid filtration coefficient （Kf） and osmotic inflation coefficient （δ） in proteins perfused with plain Hank＇s balanced salt solution, which contains 1 g/L albumin. Testing was taken on the 1st day, 3rd day, 5th day and 7th day. Western blot was also used to determine the expression of survivin on the 5th day. Results Groups responding purely to high glucose showed an increased EC monolayer permeability on the 3rd, 5th and 7th days, compared to the control group and the Ang-glucose group. This increased tendency could also be seen in the Ang-glucose group on the 5th and 7th days （P〈0.01）. Western blot showed a significant difference between the group with Ang-1 and the group without Ang-1. Conclusion High-concentration glucose increases EC monolayer permeability. The beneficial effect of Ang-1 limits this damage and may be a method for up regulating the expression of survivin to prevent RF/6A from apoptosis.
出处
《眼视光学杂志》
2009年第2期119-122,共4页
Chinese Journal of Optometry & Ophthalmology
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)
