白茶、玫瑰花及玳玳花提取物对体外培养黑素细胞功能的影响

Effects of the extracts of white tea,rose and bitter citrus immature flower on the function of melanocytes in vitro

关键词:  祛斑因子A、B  黑素细胞  角质形成细胞  作用机制

Key words:  freckle factor A and B  melanocyte  keratinocyte  mechanism

摘要:

[目的]观察白茶、玫瑰花及玳玳花提取物（祛斑因子A、B）对体外培养的正常人表皮黑素细胞功能及角质形成细胞和黑素细胞共培养体系中黑素小体转运的影响。[方法]制备含药血清，建立正常人表皮黑素细胞体外培养体系，采用4-甲基偶氮唑蓝（MTT）法测定黑素细胞增殖，酪氨酸酶多巴速率氧化法测定酪氨酸酶活性，NaOH裂解法测定黑素生成量，逆转录-聚合酶链反应（RT-PCR）检测黑素细胞酪氨酸酶mRNA的表达，Pull-down法检测黑素细胞GTP-RhoA和GTP-Rac1蛋白的表达；建立正常人角质形成细胞和黑素细胞的体外共培养体系，流式细胞术检测共培养细胞体系中黑素转运的情况。[结果]与空白血清对照组相比，祛斑因子A、B可显著抑制酪氨酸酶活性；祛斑因子A可显著降低酪氨酸酶mRNA的表达。祛斑因子A中、低剂量可抑制树突结构蛋白的表达。祛斑因子A高、中、低剂量及祛斑因子B均可抑制黑素小体的转运。[结论]为祛斑因子用于色素性疾病的保健防治提供一定的实验依据。

Abstract:

[Objective] To observe the effect of extracts of white tea,rose and bitter citrus immature flower (freckle factor A and B)on the function of normal human epidermal melanocytes and the transport of melanosomes in the co-culture system of epidermal keratinocytes and melanocytes.[Methods] Human melanocytes was cultured in vitro and the medicated serum of mice was prepared from freckle factor A and B. The proliferation of melanocytes,the tyrosinase activety,melanin synthesis,the expression of tyrosinase mRNA and the expression of GTP-RhoA and GTP-Rac1 proteinswere measured by MTT,dopaoxidase,NaOH methods respectively,RT-PCR and Pull-down assay. Flow cytometry was used to detect the melanin transport in vitro co-cultured cell system of melanocyte and epidermal keratinocytes. All of the results were compared to those treated with Taitai oral liquid and vitamin C.[Results] In freckle factor A,low dose could inhibit the expression of dendritic structural protein,medium and low dose of freckle factor A and B could inhibit the transport of melanosomes. The extracts of white tea,rose and hawksdale have inhibitory effect on tyrosinase activity of melanocytes cultured in vitro. The activity of tyrosinase was significantly inhibited by freckle factor A and B compared with the control group. Freckle factor A could significantly reduce the expression of tyrosinase mRNA.[Conclusion] This study can provide some experimental basis for the application of freckle factor A and B in the prevention and treatment of pigmented diseases.