| ?-glucan as an important material for modern cosmetics,commonly used in moisturizing and anti-wrinkling.In this study,B16 / HaCat was used as the cell model to research the whitening effect and whitening mechanism of the ?-glucan from Agaricus bisporus.In vitro chemical experiments,the ?-glucan from Agaricus bisporus had a free radical scavenging effect in a certain concentration range but not significant,suggesting that it is meaningless to study the whitening effect from antioxidant pathway.In the inhibition experiment of mushroom tyrosinase activity,?-glucan and arbutin had a significant inhibitory effect on mushroom tyrosinase activity in the same concentration range(0 ? 100 ?g / ml)and positively correlated with the concentration of action.The results showed that the ?-glucan had no significant effect on the cell proliferation,cell morphology and cell cycle of B16 cells at 0 ? 2000 ?g / ml,while the high concentration of arbutin(100 ?g / ml)had significant effect(P < 0.05);When the ?-glucan's concentration was 500 ?g / ml or 2000 ?g / ml,the activity of tyrosinase in B16 cells was significantly lower than that in the control group(P < 0.001)and better than arbutin's(100 ?g / ml)effect;and it was found that ?-glucan could inhibit the synthesis of melanin in B16 cells by 8-MOP.Through RT-PCR and Western Blot,?-glucan on down-regulation m RNA level of Tyrosinase,TRP 1 and TRP 2,which were not significantly different,but the protein expression level was significantly down-regulated.The results showed that the ?-glucan could down-regulation the protein expression level of B16 cells produce melanin-related enzymes Tyrosinase,TRP 1 and TRP 2,further explained the whitening mechanism of ?-glucan.B16 / HaCat co-culture cells showed that there was no significant effect on the growth sequence of co-cultured cells.When the quantity of B16 and HaCat cells was 1:4,the two kinds of cells were grown well.Then on the co-culture model we could research the effects of UV irradiation and the role of ?-glucan.The results showed that ?-glucan and arbutin had a certain inhibitory effect on tyrosinase activity and intracellular melanin content on co-cultured cells.It was also found that under high concentration of ?-glucan,the content of melanin in B16 cells was more than that in HaCat cells,while the content of melanin in HaCat cells was higher than that in B16 cells of arbutin.The results showed that ?-glucan had more obvious inhibition of melanin from B16 cells to HaCat cells metastasis than arbutin. |
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