Department of Dermatology, Affiliated Hospital of Jiangnan University, Wuxi, 214062, Jiangsu, China.
Department of Pathology and Laboratory Medicine, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, 19104, USA.
Lasers Med Sci. 2021 Feb;36(1):139-146. doi: 10.1007/s10103-020-03047-6. Epub 2020 Jun 30.
Phototherapy is an effective therapeutic option in the treatment of vitiligo; however, responses varied among the different types. The underlying mechanism has scarcely been investigated. To investigate and compare the effects of phototherapy on the mutation of melanocyte lineage differentiated from human scalp-derived neural crest stem cells (HS-NCSCs) with p75 neurotrophin receptor expression positive and p75 neurotrophin receptor expression negative group in vitro, the HS-NCSCs were isolated from fetal scalp tissue, which is identified by immunofluorescent staining. The p75(+) and p75(-) cells from HS-NCSCs were isolated by magnetic cell sorting, respectively. The embryonic neural crest stem cell biomarkers were detected by RT-PCR. Narrow-band UVB (NB-UVB) was used to irradiate the cells. Cell proliferation was evaluated by cell count. Tyrosinase, Tyrp1, and Tyrp2 gene expression were measured by quantitative RT-PCR. Tyrosinase and GRCR protein levels were investigated by Western blot analysis. The electrophoretic strip showed that Sox2, Oct4, Sox10, and Nestin of p75(+) HS-NCSCs were brighter than the p75(-) HS-NCSCs. After the same dose radiation with NB-UVB, the cell proliferation of p75(+) group showed less inhibitory rate compared with the p75(-) HS-NCSCs. The tyrosinase mRNA and protein expression of differentiated melanocytes increased significantly in the group of p75(+) HS-NCSCs compared with the p75(-) group. The melanocytic mutation of p75(+) HS-NCSCs increased significantly compared with the p75(-) HS-NCSCs under NB-UVB, which indicated there were more melanocyte precursors in the differentiated cells from p75(+) HS-NCSCs. This may provide new insights for the different repigmentation efficacy of segmental and non-segmental vitiligo.
光疗是治疗白癜风的有效治疗选择；然而，不同类型的反应有所不同。其潜在机制尚未得到充分研究。为了研究和比较光疗对体外表达 p75 神经生长因子受体阳性和 p75 神经生长因子受体阴性人头皮源性神经嵴干细胞（HS-NCSCs）黑素细胞系分化突变的影响，从胎头皮组织中分离 HS-NCSCs ，通过免疫荧光染色进行鉴定。通过磁细胞分选分别分离 p75(+)和 p75(-)细胞。通过 RT-PCR 检测胚胎神经嵴干细胞标志物。窄谱 UVB（NB-UVB）用于照射细胞。通过细胞计数评估细胞增殖。通过定量 RT-PCR 测量酪氨酸酶、Tyrp1 和 Tyrp2 基因表达。通过 Western blot 分析研究酪氨酸酶和 GRCR 蛋白水平。电泳条带显示 p75(+) HS-NCSCs 的 Sox2、Oct4、Sox10 和 Nestin 比 p75(-) HS-NCSCs 更亮。经相同剂量 NB-UVB 辐射后，p75(+)组细胞增殖抑制率较 p75(-) HS-NCSCs 降低。与 p75(-)组相比，p75(+) HS-NCSCs 分化的黑素细胞酪氨酸酶 mRNA 和蛋白表达显著增加。与 p75(-) HS-NCSCs 相比，p75(+) HS-NCSCs 在 NB-UVB 下的黑素细胞突变明显增加，这表明 p75(+) HS-NCSCs 分化细胞中的黑素细胞前体更多。这可能为节段性和非节段性白癜风不同的复色效果提供新的见解。
