目的:探讨咖啡因在体外培养人毛乳头细胞中抗雄激素作用的机制。方法:体外培养人毛乳头细胞经0.0005%咖啡因及10nmol/L睾酮单独及联合处理48h后,通过碱性磷酸酶(AKP)染色、四甲基偶氮唑蓝(MTT)、流式细胞仪检测,并筛选10个基因通过RealtimePCR检测mRNA水平表达量。结果:0.0005%咖啡因能对抗睾酮对毛乳头细胞的凋亡,促进增殖;顶部毛乳头细胞经睾酮处理后雄激素受体(AR)、Ⅱ型5α-还原酶(SRDSA2)、p53基因、凋亡信号受体FasR、糖原合成酶激酶(GSK)-3β、转化生长因子(TGF)-β2表达上调,加入咖啡因后可部分逆转睾酮的生理作用。阴部毛乳头细胞睾酮处理后p53、FasR等凋亡因子的表达下降,加入咖啡因可以进一步抑制细胞凋亡。结论:咖啡因在体外培养人毛乳头细胞中可能通过多条信号转导通路发挥抗雄激素作用。
Objective: To investigate the possible antiandrogenic mechanisms of caffeine on human dermal papilla cells in vitro. Methods: Human dermal papilla cells, taken from vertex, occiput and pubis, were cultivated for 48 h in vitro with 0.0005% caffeine and 10 nmol/L testostero respectively or combination. Measured the relative growth and apoptotic rate by MTT assay and flowcytometry, and Realtime PCR was applied to analysis mRNA of 10 candidate genes connecting to possible signaling pathway of AGA. Results: 0.000 5% caffeine stimulated the proliferation and inhibited the apoptosis of human DPCs in vitro. AR, SRDSA2, p53, FasR, GSK-3β TGF-β2 show significant regulations in vertex DPC culture treatment concentra- tions of 10 nmol/L testostero. In pubic cell culture p53 and FasR were down- regulated with testostero and further apoptosis suppression can be achieved by caffeine treatment. Conclusion: Caffeine probable play a role in antiandrogenic effect on hu- man dermal papilla cellss in vitro by acting on different signalling pathway.