| ?-glucan is an important active components of whitening skin products,while the mechanism of its inhibitory activity during melanogenesis remains unclear.This subject uses zebrafish as an animal model to explore the molecular mechanism of Agaricus bisporus ?-glucan on melanogenesis inhibition.In this study we firstly proved that ?-glucan treatment could obviously inhibit the deposition of melanin on the body surface in larval and adult zebrafish through calculating and analyzing the pigmentation area.Moreover,?-glucan can still significantly inhibit massive accumulation of melanin induced by 8-MOP in vitro and overexpressing of mitfa,tyr m RNA in vivo.Nextly,we detected the transcription level of key genes during melanin formation by semi-quantitative RT-PCR and in situ hybridization.The results showed that ?-glucan treatment could inhibit tyr,tyrp1 b,dct and silv m RNA levels.In addition,the protein levels of tyrosinase(TYR),tyrosinase related protein 2(TRP2),silver(SILV)and the enzyme activity of tyrosinase were also significantly downregulated by ?-glucan through Western Blot and tyrosinase activity detection.We furtherly explored the mechanism about how ?-glucan functions in inhibiting melanin formation.?-glucan can reduce the increasement of tyr,tyrp1 a,tyrp1b and dct expression caused by overexpression of mitfa.What's more,?-glucan treatment also inhibited the expression of mc1 r in different developmental stages,suggesting that ?-glucan maybe regulates the differentiation of melanocytes by inhibiting the c AMP/PKA signaling pathway and down-regulating the expression of mitfa.Finally,the reversible effect of ?-glucan on melanin formation was verified in this study.The results showed that such inhibitory effect could be reversed after the withdrawal of ?-glucan.Indicating that the inhibitory melanin effect of ?-glucan on zebrafish was reversible.Our results showed that ?-glucan can significantly inhibit zebrafish melanogenesis,supporting the notion that ?-glucan could potentially be used in the development and production of natural whitening products. |
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