单核巨噬细胞集落刺激因子对人皮肤成纤维细胞增殖及胶原合成的影响

Effect of Granulocyte/Macrophage Colony-Stimulating Factor(GMCSF)on cell proliferation and synthesis of collage in human derma fibroblast

目的:研究单核巨噬细胞集落刺激因子(granulocyte/macrophage colony-stimulating factor,GMCSF)对人皮肤成纤维细胞增殖及胶原合成的影响,探讨其促进创面愈合的机制并为临床准确定量应用该因子促进创面愈合提供理论依据。方法:分别应用浓度为0、25、50、75、100、150ng/mlGMCSF培养液孵育离体培养的人皮肤成纤维细胞,作用24h后四甲基偶氮唑盐比色法(MTT)检测细胞的活性。选择最适浓度GMCSF干预细胞,用流式细胞仪检测成纤维细胞的周期变化;应用ELISA检测上清液中Ⅰ、Ⅲ型胶原合成情况。结果:GMCSF在浓度为25～100ng/ml之间对人皮肤成纤维细胞有明显的促增殖作用,其中以50/ml最为显著;成纤维细胞在最适浓度GMCSF干预24h后,细胞大多处于DNA合成前期和合成期;与空白组比较,GMCSF组Ⅰ、Ⅲ型胶原分泌明显增加(P<0.01),且Ⅰ、Ⅲ型胶原比值下降(P<0.01)。结论:GMCSF在浓度为25～100ng/ml对人皮肤成纤维细胞有明显的促增殖作用,且能刺激成纤维细胞合成大量Ⅰ、Ⅲ型胶原,这可能是其加速创面愈合的机制之一。

Objective To investigate the effects of Granulocyte/Macrophage Colony-Stimulating Factor（GMCSF）on cell proliferation and synthesis ofcollage in cultured human derma fibroblasts.To explore the mechanism by which GMCSF promotes wound healing and provide a theory basis for application GMCSF at the accurate quantification in the clinical.Methods In vitro human derma fibroblasts in good status were incubated in different concentration（0、25、50、75、100、 150ng/ml） of GMCSF culture medium.After 24 hours,cel1 activity was detected with MTT.Cell cycle of fibroblasts interfered with the best concentration were measured by flow cytometry.And the secretion of typeⅠand Ⅲ collagen in supernatant was measured by enzyme linked immunosorbant assay（ELISA）.Results GMCSF is effective to induce the proliferation of fibroblasts between 25ng/ml to 100ng/ml,especially at the concentration 50ng/ml.The fibroblasts interfered with optimum concentration for 24 hours were in pre-and synthetic DNA synthesis period.Compared with the blank group,the supernatant of the type Ⅰand Ⅲ collagen were raised remarkably in GMCSF group（P〈0.01） and decreased ratio of the type Ⅰ to the type Ⅲ collagen（P〈0.01）.Conclusion GMCSF is effective to induce the proliferation of fibroblasts between 25ng/ml to 100ng/ml and can up-regulate directly the expression of typeⅠand Ⅲ collagen in human derma fibroblast,which may be one of the mechanisms that GMCSF accelerates the wound healing.