氨甲环酸纳米乳通过诱导细胞自噬抑制黄褐斑的作用机制研究

Mechanism of Tranexamic Acid Nanoemulsion in the treatment of Melasma through Autophagy pathway

关键词:  氨甲环酸  细胞自噬  纳米乳  黄褐斑  黑色素

Key words:  tranexamic acid  autophagy  nanoemulsion  melasma  melanin

摘要:

目的 验证氨甲环酸（TXA）纳米乳是否通过细胞自噬途径影响黄褐斑的发生与发展。方法 采用不同浓度的TXA纳米乳处理B16细胞（黑色素瘤），以MTT法检测细胞增殖情况，NaOH裂解法检测蛋白质黑色素含量，L-Dopa氧化法测定细胞内酪氨酸酶活性，并通过免疫荧光法和Western blot法检测细胞相关蛋白的表达情况。结果 MTT结果显示不同浓度TXA纳米乳对B16细胞具有增殖抑制作用，且TXA纳米乳可显著降低黑色素的生成、抑制细胞内酪氨酸酶活性。Western blot结果显示，TXA纳米乳可显著下调黑色素生成蛋白（MIFF、TRP1/2）的表达，上调自噬相关标记蛋白（Beclin1、ATG12、LC3）的表达。免疫荧光法显示TXA纳米乳可以上调LC3蛋白在B16细胞胞质中的表达。结论 TXA纳米乳通过激活细胞自噬系统影响相关蛋白的表达，减少黑色素沉着，从而实现黄褐斑的有效治疗。

Abstract:

OBJECTIVE To verify whether Tranexamic acid(TXA) nano-emulsion affects the occurrence and development of melasma through autophagy pathway . METHODS B16 cell were treated with TXA nanoemulsion at different concentrations. The cell proliferation, melanin content, tyrosinase activity and the expression of cell-related proteins were detected by the MTT assay, NaOH lysis method, L-Dopa oxidation method, and immunofluorescence and Western blotting. RESULTS The results of MTT showed that TXA nanoemulsion at different concentrations could inhibit the growth of B16 cells, and significantly reduce melanin production and inhibit intracellular tyrosinase activity. Western blotting results showed that TXA nanoemulsion could down-regulate the expression of melanin-producing proteins (MIFF, TRP1/2) and up-regulate the expression of autophagy-related marker proteins (Beclin1, ATG12, LC3). Immunofluorescence observation showed that TXA nanoemulsion could up-regulate the expression of LC3 protein in the cytoplasm of B16 cells. CONCLUSION TXA nanoemulsion can affect the expression of related proteins by activating the autophagy system of cells and reduce melanosis, so as to achieve effective treatment of melasma.