N-tert-Butylhydroxylamine promotes melanin production in oxidative stress conditions through the MITF signaling pathway.

During the aging process, melanogenesis is downregulated, leading to hair graying. Various agents, including α-melanocyte-stimulating hormone (α-MSH), have been evaluated for their potential to enhance melanogenesis. Among them, N-tert-Butylhydroxylamine (NtBHA) has been reported to reverse cellular aging, enhance mitochondrial function, and exhibit antioxidant properties. The effect of NtBHA on melanogenesis remains unclear. This study aimed to explore the impact of NtBHA on melanogenesis in B16F1 cells. In this study, NtBHA exhibited strong antioxidant activity, as demonstrated by antioxidant activity, lipid peroxidation assay, and hydroxyl radical assay. Importantly, NtBHA did not show any cytotoxic effects in the MTT assay. Additionally, in a melanin production experiment in the presence of H₂O₂, NtBHA increased melanin production in cells exposed to oxidative stress induced by H₂O₂. Additionally, NtBHA enhanced the expression of crucial proteins related to melanogenesis, such as MITF, TRP-2, and TYR. Therefore, NtBHA may promote melanogenesis in H₂O₂-stressed melanoma cells, which are aged, potentially supporting melanogenesis through the MITF signaling pathway. In addition to its potential role in restoring pigmentation, the robust antioxidant properties of NtBHA underscore its promise as a therapeutic agent for managing pigmentation disorders and mitigating oxidative stress-related skin aging. This study establishes a foundation for further research into the clinical application of NtBHA in anti-aging and dermatological treatments.