摘要
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 prove that fibroblasts could produce and release the keratinocyte growth factor which can promote epidermal cell proliferation. Methods Fibroblasts and epidermal cells obtained from prepuces were separately cultured. The transcription level of KGF in fibroblasts was measured by RT-PCR. ELISA was used to detect the level of KGF in the supernatant of fibroblast culture solution,which was used to culture epidermal cells. And then the cells were measured by cytometry,MTT assays and the immunofluorescence method. Results The results of RT-PCR showed a high transcription level of KGF in fibroblasts. A high level of KGF was detected in the supernatant of fibroblast culture solution by ELISA. The results of cytometry and MTT assays showed that the supernatant of fibroblast culture solution could promote epidermal cell proliferation and keep their activities. The test of immunofluorescence proved that KGF in the supernatant of fibroblast culture solution could combine with the specific receptors on the surface of epidermal cells. Conclusion Fibroblasts could synthesize and release the keratinocyte growth factor,which could be one of the main factors to promote epidermal cell proliferation.
出处
《山东大学学报（医学版）》
CAS
北大核心
2010年第4期18-22,共5页
Journal of Shandong University:Health Sciences
基金
国家自然科学基金资助课题(30670571
30772258)
山东省教育厅科研发展计划资助项目(J07WD03)
国家重点基础研究发展计划(973计划)(2005CB522603)
关键词
成纤维细胞
表皮细胞
角质细胞生长因子
增殖
Fibroblasts
Epidermal cells
Keratinocyte growth factor
Proliferation
