Abstract

Chlorogenic acid (CGA), the ester formed between caffeic acid and l-quinic acid, is a widespread phenolic compound. It is part of the human diet, found in foods such as coffee, apples, pears, etc. CGA is also was widely used in cosmetics, but the effects of CGA on melanogenesis are unknown. In this study, we analyzed the effects of CGA on cell proliferation, melanin content and tyrosinase of B16 murine melanoma cells. Additionally, the enzymatic reactions of CGA in B16 melanoma cells lytic solution were detected by UV spectrophotometry. Results showed CGA at 30 and 60 μM significantly suppresses cell proliferation. 8-MOP at 100 μM significantly promotes cell proliferation, but CGA can counter this. Incubated for 24 h, CGA (500 μM) improves melanogenesis while suppressing tyrosinase activity in B16 melanoma cells or 8-methoxypsoralen (8-MOP) co-incubated B16 melanoma cells. After 12 h, B16 melanoma cell treatment with CGA leads to an increase in melanin accumulation, however, after 48 h there is a decrease in melanin production which correlates broadly with a decrease in tyrosinase activity. CGA incubated with lytic solution 24 h turned brown at 37 °C. The formation of new products (with a maximum absorption at 295 nm) is associated with reduction of CGA (maximum absorption at 326 nm). Therefore, CGA has its two sidesroles in melanogenesis of B16 melanoma cells. CGA is a likely a substrate of melanin, but the metabolic product(s) of CGA may suppress melanogenesis in B16 melanoma cells by inhibiting tyrosinase activity.

Highlights

  • Phenolic compounds are common in all fruits and vegetables

  • We evaluated the effects of Chlorogenic acid (CGA) on the growth of B16 melanoma and 8-MOP-treated B16 melanoma cells

  • The cell viability with 8-MOP at 100 μM was significantly different from the control

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Summary

Introduction

Phenolic compounds are common in all fruits and vegetables. Chlorogenic acid (CGA) is the most important cinnamic acid derivative and accumulates to high levels in some crop plants, such as coffee, apples, pears, etc. CGA is an important bioactive compound, abundant in some Traditional Chinese. The antioxidant activities of CGA are due to inhibition of the formation of reactive oxygen species (ROS) or by scavenging them. UV light activates tyrosinase by microphthalmia-associated transcription factor (MITF) [12]. 8-MOP stimulates levels of microphthalmia-associated transcription factor (MITF) expression via PKA pathway, which in turn stimulates tyrosinase expression [15]. CGA is a substrate of PPO in browning reactions, but there is little data about the effect of CGA on melanocytes. This prompted us to investigate the potential of CGA to treat hypopigmentation (characterized by a loss of skin pigmentation) or hyperpigmentation (recognized by the presence of dark plaques on the skin)

Effects of CGA on the Proliferation of B16 Melanoma Cells
Effects of CGA on Melanin Production and Tyrosinase Activity
Effects of CGA on Cells Lysis Solution Enzymatic Reaction Assay
Materials
Cell Culture and MTT Assay
Tyrosinase Activity
Melanin Content
Enzymatic Reaction
General Procedures
Conclusions
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