芹菜素-7-葡萄糖苷、根皮素和柚皮素对 B16F10 黑素瘤细胞酪氨酸酶活性和黑色素合成的影响。

Effect of apigenin-7-glucoside, genkwanin and naringenin on tyrosinase activity and melanin synthesis in B16F10 melanoma cells.

机构信息

Unit of Bioactive and Natural Substances and Biotechnology UR12ES12, Faculty of Pharmacy, University of Monastir, Avicenne Street, Monastir 5000, Tunisia; Laboratory of Cellular and Molecular Biology, Faculty of Dental Medicine, University of Monastir, Avicenne Street, Monastir 5000, Tunisia.

出版信息

Life Sci. 2016 Jan 1;144:80-5. doi: 10.1016/j.lfs.2015.11.030. Epub 2015 Nov 30.

DOI:10.1016/j.lfs.2015.11.030

PMID:26656314

Abstract

AIMS

In this study, we have investigated the effects of apigenin-7-glucoside, genkwanin and naringenin, on mouse melanoma B16F10 cell proliferation. Influence of these natural products on percentage cell distribution in cycle phases and melanogenesis was also studied.

MAIN METHODS

Cell viability was determined at various periods using the MTT assay, whereas effects of tested compounds on progression through the cell cycle were analyzed by flow cytometry. In addition, amounts of melanin and tyrosinase were measured spectrophotometrically at 475 nm. Besides, the mechanism involved on the death route induced by the tested molecules was evaluated using the bis-benzimide trihydrochloride coloration method (Hoechst 33258).

KEY FINDINGS

Apigenin-7-glucoside, genkwanin and naringenin exhibited significant anti-proliferative activity against B16F10 melanoma cells after 24 and 48 h of incubation. Furthermore, apigenin-7-glucoside, genkwanin and naringenin provoked an increase of subG0/G1, S and G2/M phase cell proportion with a significant decrease of cell proportion in G0/G1 phases. The results evaluated using Hoechst 33,258, confirm that the percentage of B16F10 cells observed in the sub G0/G1 phase were undergoing apoptosis. Moreover, apigenin-7-glucoside and naringenin revealed an ability to enhance melanogenesis synthesis and tyrosinase activity of B16F10 melanoma cells. Whereas genkwanin induces a decrease of melanin synthesis by inhibiting tyrosinase activity.

SIGNIFICANCE

Our results promote the introduction of genkwanin in cosmetic preparations, as skin whitening agent, whereas apigenin-7-glucoside and naringenin should be introduced into cosmetic products as natural tanning agents.

摘要

目的

本研究旨在探讨芹菜素-7-葡萄糖苷、金纳多和柚皮苷对小鼠黑素瘤 B16F10 细胞增殖的影响。还研究了这些天然产物对细胞周期各时相分布和黑色素生成的影响。

方法

采用 MTT 法在不同时间点测定细胞活力，用流式细胞术分析受试化合物对细胞周期进程的影响。此外，用分光光度法在 475nm 处测量黑色素和酪氨酸酶的含量。此外，还通过双苯并咪唑三盐酸盐染色法（Hoechst 33258）评估了测试分子诱导的死亡途径的机制。

主要发现

经过 24 和 48 小时孵育，芹菜素-7-葡萄糖苷、金纳多和柚皮苷对 B16F10 黑色素瘤细胞表现出显著的抗增殖活性。此外，芹菜素-7-葡萄糖苷、金纳多和柚皮苷可增加亚 G0/G1、S 和 G2/M 期细胞比例，显著降低 G0/G1 期细胞比例。Hoechst 33258 的结果证实，观察到的 B16F10 细胞亚 G0/G1 期比例正在经历细胞凋亡。此外，芹菜素-7-葡萄糖苷和柚皮苷增强了 B16F10 黑色素瘤细胞的黑色素合成和酪氨酸酶活性。而金纳多通过抑制酪氨酸酶活性，导致黑色素合成减少。