高糖诱导人脐静脉内皮细胞中玻连蛋白的表达和细胞骨架重塑  

Vitronectin expression and cytoskeleton remodeling in human umbilical vein endothelial cells induced by high glucose

作　　者：王玉风[1,2] 杨侠[1,2] 董晓光[1,2] 

机构地区：[1]青岛大学 [2]山东省眼科研究所,山东省青岛市266071

出　　处：《眼科新进展》2012年第6期512-516,共5页Recent Advances in Ophthalmology

基　　金：山东省自然科学基金资助(编号:ZR2011HM012);青岛市科技计划基础研究项目(编号:10-3-4-13-3-JCH)~~

摘　　要：目的观察高糖培养的人脐静脉内皮细胞(human umbilical vein endothelial cells,HUVEC)中玻连蛋白(vitronectin,VN)表达的变化、细胞微丝骨架的改变以及细胞迁移能力、成管能力的变化。方法建立高糖培养的HUVEC体外模型,并常规培养HUVEC作为正常对照,Western blot方法检测VN蛋白的表达,免疫荧光细胞化学法观察HUVEC中微丝骨架F-actin的变化,划痕法检测HUVEC迁移能力的改变,Matrigel法检测HUVEC的成管能力。结果培养24h时,正常组和高糖组中VN蛋白表达量分别为0.893±0.119、1.295±0.080,48h时分别为0.874±0.115、1.724±0.098,差异均有统计学意义(t=-4.857、-9.710,均为P<0.05)。免疫荧光细胞化学结果显示培养24h、48h时高糖组中都伴有更加明显的细胞微丝骨架结构改变。划痕法检测培养24h时正常组和高糖组中迁移到划痕内的细胞数分别为(198.000±15.880)个、(242.000±11.130)个,48h时分别为(293.000±8.681)个、(334.000±10.863)个,差异均有统计学意义(t=-5.495、-7.193,均为P<0.05)。Matrigel法发现培养6h时高糖组中形成的闭合管腔数目(120.000±9.813)个多于正常组(91.000±9.867)个,12h时形成的闭合管腔数目(19.000±3.209)个也多于正常组(8.000±3.011)个,差异均有统计学意义(t=-5.163、-5.659,均为P<0.05)。结论 VN可能在糖尿病视网膜病变中起一定作用,且VN功能的发挥可能涉及到细胞骨架重塑,但需要进一步干扰VN的表达以证实其作用。Objective To observe the changes of vitronectin （VN） expression, cytoskeleton remodeling, cell migration and blood vessels forming in human umbilical vein endothelial cells （HUVEC） cultured in high glucose. Methods HUVEC were cul- tured respectively by normal and high glucose. VN protein expression was measured by Western blot, and the microfilament cytoskeleton F-actin in HUVEC was observed by immunofluorescence staining combined with fluorescence microscope. The cell migra- tion ability was evaluated with scratch wound assay, and blood vessels forming ability was observed with Matrigel assay. Results VN protein expressions in normal and high glucose were 0. 893 ± 0. 119,1. 295 ± 0. 080 at 24 hours, and 0. 874 ± 0. 115,1. 724 ± 0.098 at 48 hours,respectively, there were statistical differences（ t = -4. 857, -9.710, both P 〈 0.05 ）. Immunofluorescence showed that cytoskeleton remodeling was obvious in high glucose at 24 hours and 48 hours. In scratch wound assay, the number of migra- ting cells in normal and high glucose were 198.000 _± 15. 880,242.000 ± 11. 130 at 24 hours ,and 293.000 ± 8.681,334.000 ± 10.863 at 48 hours ,respectively,there were statis- tical differences （ t = - 5. 495, - 7.193, both P 〈 0.05 ）. In Matrigel assay, the number of blood vessels in high glucose （ 120. 000 ± 9. 813 ） was much more than that in normal （91. 000 ±9.867） at 6 hours,and the number of blood vessels in high glucose （ 19.000 ± 3. 209 ） was also much more than that in normal （ 8. 000 ± 3.011 ） at 12 hours ,there were statistical differences（t= -5. 163, -5.659,bothP〈0.05）. Conclusion VN may play the role in diabetic retinopathy, and the functions of VN may be involve the cytoskeleton remodeling, but its expression should be further interfered to confirm its role.

关 键 词：高糖 人脐静脉内皮细胞 玻连蛋白 细胞骨架 细胞迁移 

分 类 号：R285.5[医药卫生—中药学]