Comparative Study of High-throughput Screening Models for Anti-hyperpigmentation Compounds

Abstract

The mushroom tyrosinase assay, B16-F10 mouse melanoma cell model, and zebrafish model are frequently used for high-throughput screening and are widely used for developing anti-hyperpigmentation compounds, although these systems cannot be compared. We used each of these three systems to evaluate the seven anti-hyperpigmentation compounds. We investigated 1. tyrosinase activity using a mushroom tyrosinase assay, 2. viability, tyrosinase activity, and melanin content in B16-F10 cells, and 3. embryonic toxicity, tyrosinase activity, and melanin content in zebrafish. α-Arbutin, raspberry ketone (RK), raspberry ketone glucoside (RKG), glabridin (GLA), and 3-o-ethyl-ascorbic (EA), inhibited the activity of mushroom tyrosinase; dipotassium glycyrrhizinate (DG) did not inhibit mushroom tyrosinase activity, and glycyrrhetic acid (GA) promoted tyrosinase activity. Tyrosinase activity was inhibited by α-arbutin, GLA, GA and DG in B16-F10 cells; RK, RKG and EA did not inhibit tyrosinase activity. α-Arbutin, RK, RKG, EA, and GA, inhibited tyrosinase activity in zebrafish; GLA and DG did not inhibit tyrosinase activity. α-arbutin, RK, RKG, EA, GLA, and DG reduced melanin synthesis in B16-F10 cells in a dose-dependent manner without significant toxicity; GA did not inhibit melanin synthesis. α-arbutin, RK, RKG and GA significantly reduced melanin content on the zebrafish body surface. Mushroom tyrosinase analysis was the most practical assay among the three HIGHLIGHTS• It is the first time to compare mushroom tyrosinase model, B16 mouse melanoma cell model and zebrafish model.• A high-throughput method for the quantification of melanin in zebrafish skin based on the Phyton for the first time.