Granulocyte/macrophage colony-stimulating factor treatment of human chronic ulcers promotes angiogenesis associated with de novo vascular endothelial growth factor transcription in the ulcer bed
Granulocyte/macrophage colony-stimulating factor treatment of human chronic ulcers promotes angiogenesis associated with de novo vascular endothelial growth factor transcription in the ulcer bed
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DOI：
10.1111/j.1365-2133.2005.06925.x
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发表时间：
2006-01-01
影响因子：
10.3
通讯作者：
Odorisio, T
中科院分区：
文献类型：
--
作者：
Cianfarani, F;Tommasi, R;Odorisio, T
背景粒细胞/巨噬细胞集落刺激因子（GM-CSF）是一种具有多效功能的细胞因子，已成功用于治疗慢性皮肤溃疡。 GM-CSF 对伤口愈合受损的生物学作用尚未得到部分阐明。 目的 探讨 GM-CSF 治疗慢性静脉性溃疡对病变血管化以及血管生成因子血管内皮生长因子 (VEGF) 和胎盘生长因子 (PlGF) 局部合成的影响。在给药前和给药后5天在溃疡边缘进行活组织检查。使用抗血小板内皮细胞粘附分子-1/CD31和抗α-平滑肌肌动蛋白抗体通过免疫组织化学分析伤口血管化。通过原位杂交评估 VEGF 和 PlGF 转录。为了鉴定溃疡床内转录 VEGF 的细胞群，将 VEGF 杂交信号与连续切片上不同细胞类型标记的免疫染色相关联。在经 GM-CSF 处理的培养巨噬细胞和角质形成细胞中研究了 GM-CSF 直接诱导 VEGF 转录。结果 GM-CSF 处理后溃疡床中的血管密度显着增加。在 GM-CSF 治疗之前和之后，VEGF 转录物都位于溃疡边缘的角质形成细胞中，而 VEGF 杂交信号仅在施用后在溃疡床内才明显。在溃疡边缘的角质形成细胞中几乎检测不到 PlGF mRNA，并且治疗后没有明显增加。与 VEGF 不同，在施用 GM-CSF 后，在溃疡内的细胞中无法检测到特定的 PlGF 杂交信号。单核细胞/巨噬细胞是GM-CSF处理后转录VEGF的主要细胞群。体外分析表明，VEGF 转录可以在分化的单核细胞系中被 GM-CSF 直接刺激，但在角质形成细胞中则不然。结论我们的数据表明，血管化增加与 GM-CSF 治疗慢性静脉溃疡相关，并表明炎症细胞衍生的 VEGF 可能作为 GM-CSF 在慢性溃疡中愈合作用的血管生成介质。
Background Granulocyte/macrophage colony-stimulating factor (GM-CSF), a cytokine with pleiotropic functions, has been successfully employed in the treatment of chronic skin ulcers. The biological effects underlying GM-CSF action in impaired wound healing have been only partly clarified.Objectives To investigate the effects of GM-CSF treatment of chronic venous ulcers on lesion vascularization and on the local synthesis of the angiogenic factors vascular endothelial growth factor (VEGF) and placenta growth factor (PlGF).Methods Patients with nonhealing venous leg ulcers were treated with intradermal injection of recombinant human GM-CSF, and biopsies were taken at the ulcer margin before and 5 days after administration. Wound vascularization was analysed by immunohistochemistry using antiplatelet endothelial cell adhesion molecule-1/CD31 and anti-alpha-smooth muscle actin antibodies. VEGF and PlGF transcription was assessed by in situ hybridization. To identify the cell populations transcribing VEGF within the ulcer bed, the VEGF hybridization signal was correlated with the immunostaining for different cell type markers on serial sections. Direct induction of VEGF transcription by GM-CSF was investigated in GM-CSF-treated cultured macrophages and keratinocytes.Results Blood vessel density was significantly increased in the ulcer bed following GM-CSF treatment. VEGF transcripts were localized in keratinocytes at the ulcer margin both before and after GM-CSF treatment, whereas a VEGF hybridization signal was evident within the ulcer bed only following administration. PlGF mRNA was barely detectable in keratinocytes at the ulcer margin and was not visibly increased after treatment. Unlike VEGF, a specific PlGF hybridization signal could not be detected in cells within the ulcer following GM-CSF administration. Monocytes/macrophages were the main cell population transcribing VEGF after GM-CSF treatment. In vitro analysis demonstrated that VEGF transcription can be directly stimulated by GM-CSF in a differentiated monocytic cell line, but not in keratinocytes.Conclusions Our data show that increased vascularization is associated with GM-CSF treatment of chronic venous ulcers and indicate that inflammatory cell-derived VEGF may act as an angiogenic mediator of the healing effect of GM-CSF in chronic ulcers.