Abstract
UVB light can generate potentially harmful hydrogen peroxide (H(2)O(2)) in vivo, but it can also promote the beneficial proliferation and migration of melanocytes. The successful use of UVB monotherapy for treatment of vitiligo suggests that H(2)O(2) may have a biphasic effect on melanin synthesis and melanosome transfer. To study the beneficial role of H(2)O(2) on melanogenesis and melanosome transport in living melanocytes and keratinocytes. A co-culture system model was constructed using the primary human melanocytes and keratinocytes. The MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was used to determine cell proliferation, NaOH was used to determine the melanin content, and real-time PCR was used to determine tyrosinase expression. Western blot was used to determine Rab-27A and protease-activated receptor 2 (PAR-2) expression. This study demonstrated that tyrosinase was activated by low concentrations of H(2)O(2) (≤0.3 mM); however, this activity was downregulated by high concentrations of H(2)O(2) (>0.3 mM). Activation of high levels of melanin synthesis was induced when cells were treated with low concentrations of H(2)O(2) (0.3 mM). Further observation using an in vitro co-culture system of f...Continue Reading
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