目的:白癜风是一种以黑色素细胞进行性丧失为特征的慢性皮肤病,传统疗法的疗效有限。本研究旨在建立一种易于操作的白癜风模型,高重复性,为研究白癜风的发病机制和开发新的治疗方法奠定基础。
方法:(1)建立白癜风模型:通过皮内注射将B16F10细胞输送到C57BL/6J的背部皮肤(第0天),并在第4天和第10天腹膜内注射CD4耗竭抗体。其次,在第12天手术切除黑色素瘤.第三,每第四天腹膜内施用CD8抗体直至第30天。(2)白癜风模型的鉴定:H&E染色,免疫组织化学,免疫荧光法检测黑素细胞。通过透射电子显微镜(TEM)检测黑色素,Lillie硫酸亚铁染色和L-DOPA染色。
结果:(1)背部皮肤和毛发在第30天开始出现白色。黑色素丢失在第60天达到峰值;(2)苏木精和伊红(H&E)染色,免疫组织化学和免疫荧光结果显示黑素细胞减少。L-DOPA染色,Lillie硫酸亚铁染色和TEM结果显示表皮中黑色素减少。
结论:成功建立的白癜风小鼠模型能够较好地模拟人类白癜风的发病机制,为白癜风的发病机制和治疗研究提供了重要依据。
OBJECTIVE: Vitiligo is a chronic dermatological condition characterized by the progressive loss of melanocytes, for which traditional therapy has shown limited efficacy. This study aimed to establish a vitiligo model with easy operability, high repeatability, and stable depigmentation to provide a foundation for studying the pathogenesis and developing novel therapies for vitiligo.
METHODS: (1) Establishing vitiligo model: Firstly, deliver B16F10 cells to the back skin of C57BL/6 J via intradermal injection (day 0), and the CD4 depletion antibody was injected intraperitoneally on day 4 and 10. Secondly, the melanoma was surgically removed on day 12. Thirdly, CD8 antibody was administered intraperitoneally every fourth day till day 30. (2) Identification of vitiligo model: H&E staining, immunohistochemistry, and immunofluorescence were used to detect the melanocytes. The melanin was detected by transmission electron microscopy (TEM), Lillie ferrous sulfate staining and L-DOPA staining.
RESULTS: (1) The back skin and hair began to appear white on day 30. Melanin loss reached peak on day 60; (2) Hematoxylin and eosin (H&E) staining, immunohistochemistry and immunofluorescence results showed melanocytes were reduced. L-DOPA staining, Lillie ferrous sulfate staining and TEM results showed that melanin decreased in the epidermis.
CONCLUSIONS: We successfully establishment a vitiligo mouse model which can be more capable to simulate the pathogenesis of human vitiligo and provide an important basis for the study of pathogenesis and therapy of vitiligo.