小鼠急性皮肤缺损创面粒-巨噬细胞集落刺激因子表达特点及其作用机制  

Expression and mechanism of granulocyte-macrophage colony stimulating factor in acute skin defect of the mice

作　　者：郭敏[1] 崔文慧[1] 徐祥[2] 简宇[1] 代卉[1] 杨永华 蒋建新[2] 黄宏[2] 简华刚[1] 

机构地区：[1]重庆医科大学第二附属医院创伤烧伤科,400016 [2]第三军医大学附属大坪医院野战外科研究所 创伤、烧伤与复合伤国家重点实验室 [3]长春金赛药业公司

出　　处：《中华创伤杂志》2011年第7期648-653,共6页Chinese Journal of Trauma

基　　金：国家重点基础研究发展规划资助项目（2005CB522603）;金扶宁（外用rhGM-CSF凝胶）外用吸收率及促进创而新生血管形成分横向课题资助项目（20095-20125）

摘　　要：目的探讨创面愈合过程中粒-巨噬细胞集落刺激因子（granulocyte—macrophage colony stimulating factor，GM—CSF）的表达变化及其在创面愈合中的作用和机制。方法采用小鼠全层皮肤缺损伤模型。肌肉麻醉后在背部中线近颈侧制作1．0cm×1．0cm大小皮肤缺损伤创面。50只雄性小鼠创面造模成功后，单笼饲养，将每笼编号，依次为1～50号，按完全随机原则将小鼠分为对照组（25只）和GM—CSF治疗组（25只），每组分为5个时相点，分别为5只。治疗组创面用重组GM—CSF（rhGM—CSF）凝胶（10μg／cm^2），对照组创面用凝胶基质。于伤后第3，5，7，10和14天，观察创面愈合时间；测定创面愈合率；切取创面组织，检测病理组织学变化；通过CD31免疫组化染色结果分析，计算创面微血管密度；应用RT—PCR检测创面GM—CSF、血小板源性生长因子（platelet—derived growth factor，PDGF）、血管内皮生长因子（vascular endothelial growth factor，VEGF）和基质细胞衍生因子-1（stromal cell derivedfactor-1，SDF-1）基因表达变化。结果RT—PCR检测结果显示创面GM—CSF基因表达伤后3d达峰值（P〈0．01），直到伤后10d均维持较高水平（P〈0．05），伤后14d其表达显著下降接近正常；应用rhGM—CSF凝胶后，创面愈合时间较对照组提前（2．4±0．3）d，其创面愈合率于伤后7～14d显著升高（P〈0．05）；组织学显示创伤早期创面中性粒细胞数量较少，创沿上皮细胞增殖数量较多，创面肉芽组织增生明显，并日．细胞密度大，以梭形细胞和卯圆形细胞为主，新生血管数量较多；创面微血管密度在伤后7～14d显著增加（P〈0．05）；VEGF和SDF-1基因表达，分别于伤后7d和10d内显著上调表达（P〈0．05），促愈生长因子PDGF基因于伤后各时相点均显著上调表达（P〈0．05）。结论GM—CSF在创面愈合早期表达增高，GM—CSF可促进�Objective To investigate the expression of granulocyte-macrophage colony-stimulating factor （GM-CSF） and its associated mechanism during the wound healing. Methods The animal model with the full-thickness skin injury was used in the study. Fifty male mice were involved in the study and divided randomly into control group （ n = 25 ） and GM-CSF group （ n = 25 ）. Each group had five time points （5 mice per time point）. All the mice received full-thickness skin defect （ 1 cm × 1 cm） through the panniculus carnosus on the midline of the back near the neck after anesthesia. Recombinant human granulocyte- macrophage colony stimulating factor （RhGM-CSF） gel （ 10 μg/cm2 ） were applied in the GM-CSF group and gel matrix without RhGM-CSF applied in the control group. The wound healing time and rate were observed at days 3, 5, 7, 10 and 14 after injury. The wound specimens were collected to detect the histopathological change. The microvessel density of the wound was counted based on the results of CD31 immunohistochemistry. RT-PCR was employed to detect the expression changes of GM- CSF, platelet-derived growth factor （ PDGF）, vascular endothelial,growth factor （VEGF） and stromal cell derived factor-1 （SDF-1）. Results RT-PCR results showed that the gene expression of GM-CSF reached the peak at day 3 after injury （ P 〈 0.01 ） and kept the high level at days 3-10 after injury （ P 〈 0.05 ） , followed by a sharp decrease to a normal level at day 14 after wound. The wound healing time was average （ 2.4 ± 0.3 ） days earlier than the control mice after application of rhGM-CSF, with significant in- crease of the wound healing rate during 7-14 days after injury （P 〈 0.05）. In the GM-CSF group, the early histology of trauma wound showed a small number of neutrophils, obvious epithelial cell proliferation in the wound margin, marked hyperplasia of the granulation tissue, high cell density with quantity of spin- dle-shaped and oval-shaped cells and increased number of new

关 键 词：伤口愈合 粒-巨噬细胞集落刺激因子 生长因子 基因表达 

分 类 号：R686[医药卫生—骨科学]