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目的研究成纤维细胞产生的角质细胞生长因子(KGF)对表皮细胞增殖的影响。方法自包皮分离培养成纤维细胞和表皮细胞,采用逆转录-聚合酶链反应法(RT-PCR)、酶联免疫吸附法(ELISA)、MTT法和免疫荧光等方法分别检测成纤维细胞KGF基因的转录水平、培养液中KGF的浓度、成纤维细胞培养液对表皮细胞增殖的促进作用。结果RT-PCR结果显示,成纤维细胞中KGF转录水平显著,ELISA法可检测到成纤维细胞培养液中存在较高水平的KGF,细胞计数结果和MTT结果表明,含有KGF的成纤维细胞培养液能够促进表皮细胞增殖,保持表皮细胞的活性,免疫荧光观察到KGF与表皮细胞表面的特异性受体结合。结论成纤维细胞能够合成和分泌较高水平的KGF,成为其促进表皮细胞增殖的重要因素。
Objective To study the effect of keratinocyte growth factor (KGF) produced by fibroblasts on the proliferation of epidermal cells. Methods Fibroblasts and epidermal cells were isolated and cultured from the foreskin. The expression of KGF gene in fibroblasts was detected by reverse transcription-polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), MTT and immunofluorescence The level of KGF in culture medium and the promotion effect of fibroblast culture medium on the proliferation of epidermal cells. Results The results of RT-PCR showed that the level of KGF was significantly increased in fibroblasts. The higher levels of KGF in fibroblast culture medium could be detected by ELISA. The cell counting results and MTT results showed that KGF-containing fibroblast culture medium Can promote epidermal cell proliferation, maintain the activity of epidermal cells, immunofluorescence observed KGF and epidermal cell surface receptor-specific binding. Conclusion Fibroblasts can synthesize and secrete higher levels of KGF, which is an important factor in promoting the proliferation of epidermal cells.