白消一号方作用于黑素瘤细胞与角质形成细胞共培养模型中黑素小体转运的观察

Effects of Baixiao Yihao Fang on Melanosome Transfer in the Mixed Cultivating Model of Keratinocytes and Melanoma Cells

目的体外建立黑素瘤细胞与角质形成细胞直接接触的培养模型,观察白消一号方对此模型中黑素小体转运的影响,探讨其治疗白癜风的药理作用。方法体外培养黑素瘤细胞和角质形成细胞,用二乙酰羧基荧光素-琥珀酰亚胺酯(CFDA-SE)标记黑素瘤细胞内的黑素小体,然后将标记的黑素瘤细胞与角质形成细胞共同培养。大鼠中药灌胃后取血清加入此模型中,在倒置显微镜下观察混合细胞模型生长及共聚焦显微镜下观察共培养模型中黑素小体由黑素瘤细胞向角质形成细胞的转运情况。结果含药血清可使黑素小体由黑素瘤细胞向角质形成细胞的转运增加。结论在共聚焦显微镜下观察到白消一号方可以促进黑素小体转运,此途径可能为白消一号方治疗白癜风的作用机制之一。

Objective To construct the mixed cultivating model of keratinocytes and melanoma cells, and to observe the effect of Baixiao Yihao Fang on its melanosome transfer in this model. Methods Melanoma cells and keratinocytes were cultured respectively, then the melanosomes in the melanoma cells were fluorescent labeled with carboxyfluorescein diacetate succinimidyl ester （CFDA-SE）. Finally, keratinoeytes and the labeled melanoma were co-cultured. After mouse serum containing Chinese medicine was added to the model, the growth of mixed cultivating model was observed by inverted microscopy, and melanosome transfer from melanoma to keratinocytes were observed by confocal microscopy. Results The serum of containing Chinese medicine promoted melanosome transfer from melanoma to keratinocytes. Conclusion Baixiao Yihao Fang has a strong promotion on melanosome transfer from melanoma to keratinocytes, and these results shows that Baixiao Yihao Fang could treat vitiligo through this mechanisms.