胶原海绵促进人脐静脉血管内皮细胞生长的体外实验研究

目的探讨胶原海绵促进人脐静脉血管内皮细胞生长的疗效与机制。方法实验组在细胞培养孔中加入含8 U胶原酶的0.2 ml PBS 2 mm×2 mm×2 mm正方体胶原海绵;对照组仅加入含8 U胶原酶的0.2 ml无菌的PBS液体。培养24 h后测血管内皮细胞增殖(MTT实验)、迁移(划痕实验)、及免疫蛋白印迹检测血管生成素1(Ang1)、血管内皮生长因子(VEGF)与整合素(Integraαv)。结果细胞培养后24 h,实验组可以显著促进HUVEC的增殖,增殖率达86%;实验组血管内皮细胞细胞迁移加快,12 h细胞迁移完成,显著高于对照组(P0.01);实验组VEGF、Ang1及Integraαv蛋白表达明显高于控制组,其中以整合素аv蛋白表达增加尤为显著,增加了80%。结论胶原海绵促进血管内皮细胞增殖与迁移,其中促进迁移的作用较强,机制之一为提高了血管内皮细胞中VEGF、ANG1、Integraαv蛋白表达。

Study on enhancing growth of human umbilical vascular endothelial cells with collagen sponge

Objective To explore the effect and mechanism of collagen sponge on the growth of human umbilical vascular endothelial cells. Methods All specimens were divided into experimental group and control group,specimens in experimental group were collagen sponge with cube size of 2 mm × 2 mm × 2 mm incubated with 1 ml of 8 units of collagenase loading inside indulged in the incubation fluid of HUVEC,whereas specimens in control group were indulged with only 1 ml of 8 units of collagenase. The vascular endothelial cell proliferation experiment（ MTT）,migration assay and protein expression of angiogenin 1（ Ang1）,vascular endothelial growth factor（ VEGF） and integrin integra α v with western blotting were applied on the time of 0,6,12 and 24 hours of incubation. Results The proliferation rate of HUVEC in experimental group had been increased by 86%,as compared with that of control group. The cell migration rates at 12 hours and 24 hours in experimental group were significantly higher than those of control group（ P〈0. 01）. The protein expression of VEGF,Ang1 and integra α v in experiment group was significantly higher than the results of control group with much higher protein expression of integra α v which increased by 80% in experimental group as compared with others. Conclusion It shows that positive effect on vascular endothelial cell migration and proliferation by collagen sponge with up- regulation of protein expression of VEGF,Ang1 and integra α v.