Departmet of Dermatology, Postgraduate Institute of Medical Education & Research, Chandigarh, India.
Department of Experimental Medicine & Biotechnology, Postgraduate Institute of Medical Education & Research, Chandigarh, India.
Exp Dermatol. 2019 Jun;28(6):682-689. doi: 10.1111/exd.13773. Epub 2018 Sep 19.
Cytokines regulate immune response and inflammation and play an important role in depigmentation process of an autoimmune disease, vitiligo. We sought to determine how inflammatory cytokines influence the progression of vitiligo, and based on that, we develop a logical therapeutic intervention using primary melanocyte culture. Melanocytes were cultured and exposed to IL-17A, IL-1β, IFN-γ and TGF-β for 4 days. Melanocytes proliferation, tyrosinase assay and melanin content were measured. Real-Time PCR was used to analyse mRNA expression of genes specific for melanocytes growth and pigmentation. Anti-IL-17A receptor antibody was used to block IL-17A receptors expressed on melanocytes. Protein expression of MITF and TYR was assessed by immunofluorescence and Western blotting. A gradual decline in the melanocyte population, melanin content and tyrosinase activity was observed after different cytokine treatment. The expression of MITF and its downstream genes after blocking with anti-IL-17RA, an increased melanin content, increased expression of TYR, MITF along with its downstream genes, and cell proliferation was observed.
细胞因子调节免疫反应和炎症,在自身免疫性疾病——白癜风的脱色过程中发挥重要作用。我们试图确定炎症细胞因子如何影响白癜风的进展,并在此基础上,通过原代黑素细胞培养来制定合理的治疗干预措施。培养黑素细胞并使其暴露于 IL-17A、IL-1β、IFN-γ 和 TGF-β 中 4 天。测量黑素细胞增殖、酪氨酸酶测定和黑色素含量。使用实时 PCR 分析特定于黑素细胞生长和色素沉着的基因的 mRNA 表达。使用抗 IL-17A 受体抗体阻断黑素细胞上表达的 IL-17A 受体。通过免疫荧光和 Western blot 评估 MITF 和 TYR 的蛋白表达。在不同细胞因子处理后,观察到黑素细胞群体、黑色素含量和酪氨酸酶活性逐渐下降。在用抗 IL-17RA 阻断后,观察到 MITF 及其下游基因的表达增加,黑色素含量增加,TYR、MITF 及其下游基因的表达增加,以及细胞增殖。