摘要
目的探讨骨形成蛋白-2(BMP-2),成纤维细胞生长因子-2(FGF-2)单独及联合作用对毛乳头细胞增殖和毛发诱导能力的影响。方法采用显微机械法分离大鼠触须毛乳头并培养,行免疫荧光鉴定。CCK-8法检测毛乳头细胞的增殖活性。毛发诱导能力通过检测碱性磷酸酶(ALP)活性确定。采用ELISA法检测胰岛素样生长因子-1(IGF-1)的含量。结果毛乳头细胞的ALP免疫荧光染色阳性,确定培养的细胞是毛乳头细胞。BMP-2对毛乳头细胞增殖无影响,但可提高毛乳头细胞的ALP活性并呈剂量依赖性。FGF-2促进毛乳头细胞增殖,但会降低毛乳头细胞的ALP活性并呈剂量依赖性。联合BMP-2和FGF-2既可以促进毛乳头细胞的增殖,又可以增强ALP活性,并呈时间依赖性。BMP-2可上调毛乳头细胞培养上清液中IGF-1的含量,并呈剂量依赖性。结论 BMP-2可能通过促进毛乳头细胞分泌IGF-1以保持毛乳头细胞的毛发诱导能力,按适当浓度比例联合BMP-2和FGF-2可形成互补作用,既促进毛乳头细胞的增殖,又可以保持其毛发诱导能力。
[Abstract] Objective To investigate the effects of bone morphogenetic protein-2 (BMP-2), fibroblast growth factor-2 (FGF-2) and the combination of BMP-2 and FGF-2 on the proliferation and hair inductivity of dermal papilla (DP) cells. Methods Microdissection was employed to isolate rat DP. Identification of cultured DP cells was completed by immunofluore scence staining of alkaline phosphatase (ALP). The proliferative activity of DP cells was determined by cell counting Kit-8 (CCK-8). The hair inductivity of DP cells was determined by ALP activity assay kit using microplate reader. IGF-I was assayed by enzyme linked immunosorbent assay (ELISA). Results DP cells were isolated and cultivated successfully as confirmed by cellular positive staining of ALP. BMP-2 stimulation showed no effects on DP cells proliferation but increased ALP activity of DP cells in a dose-dependent manner, while FGF-2 promoted DP cells proliferation but decreased ALP activity of DP cells. Combined stimulation of BMP-2 and FGF-2 not only promoted DP cells proliferation but also enhanced ALP activity time-dependently. BMP-2 upregulated the level of IGF-1 in culture supernates of DP cells in a dose-dependent way. Conclusion BMP-2 may maintain hair inductivity of DP cells by upregulating IGF-1, and combination of BMP-2 and FGF-2 at proper concentrations may display a mutual complement effect on the proliferation and hair inductivity of cultured DP cells.
出处
《中华临床医师杂志(电子版)》
CAS
2015年第11期105-108,共4页
Chinese Journal of Clinicians(Electronic Edition)