粒细胞-单核巨噬细胞集落刺激因子对人皮肤成纤维细胞血管内皮细胞生长因子表达的影响  

Effect of granulocyte/macrophage cohmy-stimulating factor on expression of vascular endotllelial growth factor in human dermal fibroblasts

作　　者：李晓光[1] 方勇[1] 姚敏[1] 徐鹏[1] 俞为荣[1] 倪涛[1] 王莹[1] 

机构地区：[1]上海交通大学医学院第三人民医院烧伤整形科,201900

出　　处：《中华医学美学美容杂志》2011年第4期301-305,共5页Chinese Journal of Medical Aesthetics and Cosmetology

基　　金：国家自然科学基金资助项目（编号：30872682）

摘　　要：目的探讨粒细胞-单核巨噬细胞集落刺激因子（granulocyte-macrophage colony stimulating factor，GM—CSF）对人皮肤成纤维细胞血管内皮细胞生长因子（vascular endothelial cell growth factor，VEGF）表达的影响。方法分别用含GM—CSF（GM—CSF组）和不含GM—CSF（对照组）培养液，孵育离体培养的人皮肤成纤维细胞，作用不同时间后，采用逆转录-聚合酶链反应（RTPCR）、蛋白质印迹法（Western印迹法）、酶联免疫吸附试验（EI，1SA）分别检测人皮肤成纤维细胞VEGF mRNA表达和蛋白表达。结果GM—CSF作用1、3、6、12h后，人皮肤成纤维细胞VEGF mRNA表达均较对照组显著增强（P〈0．05），其中GM—CSF作用6h后VEGFmRNA表达最强；与对照组比较，GM—CSF作用12、24、48h后。成纤维细胞内VEGF蛋白表达旺著增强，其中24h表达最强；细胞培养上清液中VEGF蛋白分泌堪随时间延长逐渐增加，各时相点GM-CSF组VEGF蛋白分泌量高于对照组（P〈0．01）。结论GM—CSF可促进人皮肤成纤维细胞VEGF的表达，这可能是GM-CSF加速创面血管新生化的机制之一。Objective To study the effects of granulocyte-macrophage colony stimulating factor GM-CSF） on the expression of vascular endotllelial growth factor （VEGF） in human dermal libroblast. Methods In vitro human dermal fibroblasts in good status were incubated with GM-CSF （GM-CSF group） or non-GM-CSF （control group） culture medium for different periods of time. The mRNA, protein expression of VEGF in derma fibroblast were determined by reverse transcription-pol- ymerase chain reaction （RT-PCR） and Western blotting, respectively, and the secretion of VEGF in supernatant was measured by enzyme linked immunosorbant assay （EIASA）. Results The expression of VEGF mRNA from dermal fihroblasts was increased significantly after 1 or more hours of incubation with GM CSF comparing with the control （P〈0.05）. 6 hours of stimulation by GM-CSF caused maxi real expression of VEGF mRNA. The expression of VEGF protein in dermal fibrobiasts was increased from 12 hours and was peaked at 24 hour after stimulation by GM-CSF. VEGF protein from the supernatant of the dermal flhroblasts was also raised persistently from 12 hour after stimulation by GM-CSF and was improved remarkably compared with the control. Conclusions GM-CSF can up-reg ulate directly the expression of VEGF in human derma fibroid）last, which may he one of the mechanisms that GM-CSF accelerates neovascularization in wound healing.

关 键 词：粒细胞-单核巨噬细胞集落刺激因子 血管内皮细胞生长因子 创面愈合 成纤 维细胞 

分 类 号：R96[医药卫生—药理学]