인간 표피 멜라닌세포/각질세포 공배양 시스템에서 유세포 분석법을 응용한 코직산의 효능 연구
Evaluation of Effect of Kojic acid by a Novel Flow Cytometric Method in Human Epidermal Melanocyte/keratinocyte Co-culture System
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Kojic acid is a fungal metabolite widely used in medicinal and cosmetic formulations as a skin whiteningagent. The activity of kojic acid is known to arise primarily from suppressing tyrosinase activity. Althoughkojic acid is effective in treating hyperpigmentation disorders in human clinical studies, it only shows weakantimelanogenic activity in mono-cultured human primary melanocytes, suggesting that there exists anotherpharmacological mechanism for in vivo activity of kojic acid. In the present study, we used primary humanepidermal melanocytes and keratinocytes co-culture system to elucidate the mechanisms of skin whiteningactivity of kojic acid. We used a newly developed flow cytometric method to analyze the counts of livemelanocytes and keratinocytes separately. When cultured without kojic acid, the proliferation of melanocytesco-cultured with keratinocytes was greatly enhanced compared to that of mono-cultured melanocytes. Onthe other hand, we found that kojic acid significantly reduced the proliferation of both melanocytes andkeratinocytes at 100~200 μM in co-culture system, although kojic acid did not reduce the proliferation ofmono-cultured melanocytes nor mono-cultured keratinocytes at the same concentration. These resultssuggest that kojic acid might suppress the expression of some growth factors produced by melanocytesor keratinocytes in co-culture system, resulting in reduced proliferation of melanocytes. In summary, themechanisms of the clinical skin whitening activity of kojic acid might include the suppression of melanocyteproliferation as well as the already known activity of tyrosinase suppression. It suggests that a cross-talkbetween melanocytes and keratinocytes is indispensable to the pharmacological mechanisms of clinicallyeffective dermatological drugs.
Abstract
Kojic acid is a fungal metabolite widely used in medicinal and cosmetic formulations as a skin whiteningagent. The activity of kojic acid is known to arise primarily from suppressing tyrosinase activity. Althoughkojic acid is effective in treating hyperpigmentation disorders in human clinical studies, it only shows weakantimelanogenic activity in mono-cultured human primary melanocytes, suggesting that there exists anotherpharmacological mechanism for in vivo activity of kojic acid. In the present study, we used primary humanepidermal melanocytes and keratinocytes co-culture system to elucidate the mechanisms of skin whiteningactivity of kojic acid. We used a newly developed flow cytometric method to analyze the counts of livemelanocytes and keratinocytes separately. When cultured without kojic acid, the proliferation of melanocytesco-cultured with keratinocytes was greatly enhanced compared to that of mono-cultured melanocytes. Onthe other hand, we found that kojic acid significantly reduced the proliferation of both melanocytes andkeratinocytes at 100~200 μM in co-culture system, although kojic acid did not reduce the proliferation ofmono-cultured melanocytes nor mono-cultured keratinocytes at the same concentration. These resultssuggest that kojic acid might suppress the expression of some growth factors produced by melanocytesor keratinocytes in co-culture system, resulting in reduced proliferation of melanocytes. In summary, themechanisms of the clinical skin whitening activity of kojic acid might include the suppression of melanocyteproliferation as well as the already known activity of tyrosinase suppression. It suggests that a cross-talkbetween melanocytes and keratinocytes is indispensable to the pharmacological mechanisms of clinicallyeffective dermatological drugs.
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