姜黄素对黑素的抑制作用及机制

The inhibitory effect of curcumin on B16F10 cells and its mechanism

目的姜黄素调节B16F10细胞微环境影响黑素合成的机制研究。方法(1)不同浓度姜黄素分别作用于B16F10及HaCaT细胞,MTT法检测该两种细胞活力;NaOH裂解法检测B16F10细胞黑素含量;L-DOPA氧化法检测B16F10细胞酪氨酸酶活性;Western blot法检测B16F10细胞黑素生成(gp100、Mitf、GPNMB、Tyr、TRP2)及转运(Rab27a)关键蛋白表达水平;HaCaT细胞:ISG15蛋白表达水平。(2)不同浓度ISG15作用于B16F10细胞,NaOH裂解法检测黑素含量;L-DOPA氧化法检测酪氨酸酶活性;结果姜黄素在一定浓度范围内,直接抑制酪氨酸酶活性及黑素生成,且抑制黑素细胞迁移;ISG15蛋白作用于黑素细胞,明显促进gp100蛋白表达,酪氨酸酶活力,黑素含量。姜黄素抑制HaCaT细胞ISG15蛋白表达,改变黑素细胞微环境,通过ISG15间接发挥抑制黑素合成作用。结论姜黄素除直接抑制黑素合成外,还可以通过抑制ISG15蛋白的表达,改变黑素细胞微环境,间接发挥抑制黑素合成作用。

Aim To investigate the effect of curcumin extract on melanin production and melanosome transport,and to explore the possible mechanism of the curcumin extract on microenvironment.Methods(1)B16F10 and HaCaT cells were cultured with different concentrations of curcumin.The proliferation ability was detected by MTT method.Melanin synthesis and tyrosinase activity in B16F10 cells were detected by NaOH pyrolysis method and Oxidation dopamine response in vitro.The expression levels of key proteins were detected by Western blot.(2)B16F10 cells were cultured with different concentrations of ISG15 protein.NaOH pyrolysis method and Oxidation dopamine response in vitro were used to detect melanin synthesis and tyrosinase activity in B16F10 cells.Results Curcumin could directly inhibited tyrosinase activity and melanin production,and inhibit melanocyte migration within a certain concentration range.ISG15 protein could enhance the melanin production,tyrosinase activity.Curcumin could reduce the expression of the ISG15 in HaCaT cell,change the microenvironment of melanocyte,and indirectly inhibit melanin synthesis through ISG15.Conclusions In addition to directly inhibiting melanin synthesis,curcumin can also play an indirect role in inhibiting melanin synthesis by inhibiting the expression of ISG15 protein and altering the microenvironment of melanocytes.