小鼠急性皮肤缺损创面GMCSF表达特点及其作用机制

发布时间：2018-05-09 20:45

本文选题：伤口愈合 + 粒-巨噬细胞集落刺激因子(GMCSF)　；参考：《重庆医科大学》2011年硕士论文

【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。局部麻醉后起效后在背部中线近颈侧制作1.0cm×1.0 cm大小全层皮肤缺损伤创面。50只雄性昆明小鼠随机分为2组:对照组(25只)和GM-CSF实验组(25只),每组分为5个时间点(每个时相点5只)。实验组创面用rhGM-CSF(recombinant human granulocyte-macrophage colony stimulating factor)凝胶(10μg/cm2)涂抹,对照组创面用凝胶基质涂抹。于伤后第3,5,7,10和14天,观察创面愈合时间;计算创面愈合率;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 derived factor-1,SDF-1)基因表达变化。结果:RT-PCR检测结果显示小鼠致创后创面GM-CSF基因表达伤后3d达峰值(P0.01),直到伤后10d均维持较高水平(P0.05),伤后14d其表达显著下调接近正常;应用rhGM-CSF凝胶后,创面愈合时间较对照组平均提前(2.4±0.27)d,其创面愈合率于伤后7～14d显著升高( P0.05);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,并且细胞密度大以梭形细胞和卵圆形细胞为主,新生血管数量较多;创面微血管密度在伤后7~14d显著增加(P0.05);VEGF和SDF-1基因分别于伤后7d和10d内表达显著上调(P 0.05),促愈生长因子PDGF基因于致伤后14d内各时相点都明显上调表达(5d,7d,10d,P0.05;14d,P0.01)。结论:GM-CSF在创面愈合早期表达增高,GM-CSF可促进创面愈合,其作用机制与PGDF、VEGF、SDF-1基因表达上调有关。
[Abstract]:Objective: to investigate the expression and mechanism of granulocyte-macrophage colony stimulating factor GM-CSF in wound healing. Methods: the model of skin deficiency injury in mice was used. After local anesthesia, 50 male Kunming mice were randomly divided into two groups: control group (n = 25) and GM-CSF group (n = 25). Each group was divided into 5 time points. Each time point is 5. The wounds in the experimental group were smeared with rhGM-CSF(recombinant human granulocyte-macrophage colony stimulating factor (10 渭 g / cm ~ 2), while the wounds in the control group were smeared with gel matrix. The wound healing time was observed, the wound healing rate was calculated, the histopathological changes were detected by excision of wound tissue, and the microvessel density of the wound was calculated by CD31 immunohistochemical staining. The expression of GM-CSF, platelet-derived growth factor-1 (PDGF), vascular endothelial growth factor (VEGF) and stromal cell derived factor-1 (SDF-1) were detected by RT-PCR. Results the results of RT-PCR showed that the expression of GM-CSF gene reached a peak value 3 days after injury in mice wound, and remained at a high level until 10 days after injury, and the expression of P0.05 mRNA was significantly down-regulated at 14 days after injury, and the expression of rhGM-CSF gel was similar to that of normal. The wound healing time was 2.4 卤0.27 days earlier than that in the control group, and the wound healing rate increased significantly at 7d after injury (P0.05). Histology showed that the number of neutrophils was less, the number of wound epithelial cells was more, and the granulation tissue proliferation was obvious in the early stage of trauma. The cell density was mainly fusiform cells and oval cells, and the number of neovascularization was more. The microvessel density of wound increased significantly within 7 and 10 days after injury, respectively, and the expression of PDGF gene increased significantly within 7 days and 10 days after injury, respectively, and the expression of PDGF gene was significantly up-regulated at all time points within 14 days after injury, including P0.0514d P0.0514d, P0.0514d, P0.0514d, P0.0514d, P0.0514d, P0.0514d, P0.0514d, P0.0514d, P0.0514d, P0.0514d, P0.0514d, respectively. Conclusion the increased expression of GM-CSF in the early stage of wound healing may promote wound healing, and the mechanism is related to the up-regulation of PGDFV EGF-1 SDF-1 gene expression.
【学位授予单位】：重庆医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R363

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