Fibrocytes from burn patients regulate the activities of fibroblasts
Fibrocytes from burn patients regulate the activities of fibroblasts
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DOI：
10.1111/j.1524-475x.2006.00192.x
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发表时间：
2007-01-01
影响因子：
2.9
通讯作者：
Tredget, Edward E.
中科院分区：
文献类型：
--
作者：
Wang, Jian Fei;Jiao, Haiyan;Tredget, Edward E.
伤口愈合需要多种细胞类型之间精心的相互作用，从而协调一系列受调控和重叠的事件。纤维细胞是参与此过程的独特白细胞亚群。这些细胞在伤口愈合中的作用之一是合成细胞外基质。有趣的是，利用质谱法定量羟脯氨酸，我们发现正常受试者或烧伤患者的纤维细胞产生胶原蛋白的能力远低于真皮成纤维细胞。因此，我们研究了纤维细胞是否可以通过影响真皮成纤维细胞的功能在烧伤创面的愈合中发挥间接的调节作用。用烧伤患者纤维细胞而不是来自正常受试者的纤维细胞调节的培养基处理真皮成纤维细胞，显示出细胞增殖和迁移的增加。使用共聚焦显微镜、流式细胞术和免疫印迹，我们发现这些处理过的真皮成纤维细胞中α-平滑肌肌动蛋白（α-SMA）的表达水平增加，这也表明收缩胶原蛋白晶格的能力增强。为了确定这些影响是否归因于转化生长因子β（TGF-β1）或结缔组织生长因子（CTGF），我们通过酶联免疫吸附测定测量了条件培养基中的总TGF-β1水平，并通过逆转录聚合酶链反应和蛋白质印迹评估了成纤维细胞和纤维细胞中CTGF mRNA和蛋白质的水平。结果显示，烧伤患者纤维细胞产生的 TGF-β1 和 CTGF 水平显着升高。此外，TGF-β1中和抗体的应用显着降低了烧伤患者纤维细胞培养基对真皮成纤维细胞增殖、迁移和胶原晶格收缩的影响。我们的结果表明，在愈合烧伤创面时，纤维细胞可以调节局部成纤维细胞的活性。
Wound healing requires an elaborate interplay between numerous cell types that orchestrate a series of regulated and overlapping events. Fibrocytes are a unique leukocyte subpopulation implicated in this process. One role proposed for these cells in wound healing is to synthesize extracellular matrix. Interestingly, using mass spectrometry to quantify hydroxyproline, we discovered that the capacity of fibrocytes from normal subjects or from burn patients to produce collagen is much less than that of dermal fibroblasts. Therefore, we investigated whether fibrocytes could play an indirect, regulatory, role in the healing of burn wounds by affecting the functions of dermal fibroblasts. Dermal fibroblasts treated with medium conditioned by burn patient fibrocytes, but not by those derived from normal subjects, showed an increase in cell proliferation and migration. Using confocal microscopy, flow cytometry, and immunoblotting, we found the level of alpha-smooth muscle actin (alpha-SMA) expression to be increased in these treated dermal fibroblasts, which also showed an enhanced ability to contract collagen lattices. To determine whether these effects could be attributed to transforming growth factor beta (TGF-beta 1) or to connective tissue growth factor (CTGF), we measured total TGF-beta 1 levels in the conditioned medium by an enzyme-linked immunosorbtion assay and assessed levels of CTGF mRNA and protein in fibroblasts and fibrocytes by reverse transcription-polymerase chain reaction and Western blotting. The results showed significantly higher levels of TGF-beta 1 and CTGF produced by burn patient fibrocytes. In addition, the application of a TGF-beta 1 neutralizing antibody significantly reduced the effect of burn patient fibrocyte medium on dermal fibroblast proliferation, migration, and collagen lattice contraction. Our results suggest that in healing burn wounds, fibrocytes could regulate the activities of local fibroblasts.