| ?-glucan had excellent biological effect,and had been widely used in the fields of medicine,food,skin care and so on.In this study,the structure of ?-glucan from Agaricus bisporus was analyzed.And its effect on cytotoxicity and the expression of cell-binding receptors of HaCat cells were investigated.Furthermore,the mechanism of UV-protection and autophagy regulation function of such ?-glucan were also addressed,thus clarify the skin care effect of ?-glucan.Which provided a scientific theoretical data for application of ?-glucan within skin care products better.The experiment was divided into three parts:?-glucan was extracted from Agaricus bisporus using alkali method,the study confirmed that ?-glucan was composed of single glucose by means of infrared spectroscopy,high performance liquid chromatography and gas chromatography.The relative molecular weight of ?-glucan was 575610.4 Da,?-glucan were glucose polymers linked together by ?-1,4glucoside bond mostly and the percentage was 86.7%,in addition,the percentage of ?-1,2glucoside bond and ?-1,6 glucoside bond was 8.38% and 4.92%.?-glucan receptor(CR3,TLR2,TLR4,TLR6 and dectin-1)gene expression of HaCat cells was tested by RT-PCR.The high concentration of ?-glucan could make the receptor gene express increasely.We had already found that HaCat cells exposed to FITC-?-glucan(?-glucan marked by FITC)for 2 h,there were 99.5% of FITC-?-glucan positive HaCat cells,in which 70.5% of the FITC-?-glucan translated from cytoplasms to nucleus by flow cytometry instrument.The results showed that the ?-glucan from Agaricus bisporus could enter HaCat cells through ?-glucan receptors on HaCat cells.UV radiation was the trigger of skin diseases,and can damage the cells.We explored the effect of ?-glucan on cell viability,cell morphology and the production of endothelin-1 of normal cells treated by UV radiation.The results showed that ?-glucan could reduce endothelin-1 production of HaCat cells,block melanin from B16 cells transfer to HaCat cells.At the same time,?-glucan had protective effects on B16 and HaCat cells treated by UV radiation,and could repair the cell injury induced by low intensity of UV radiation.We studied the effect of ?-glucan on autophagy of HaCat cells by western blot,lysosomal probe label,and immunofluorescence.The results showed that ?-glucan could promote the autophagy of normal HaCat cells,and inhibit the autophagy of HaCat cells when they are nutrient deficiency.In summary,?-glucan from agaricus bisporus could enter HaCat cells through ?-glucan receptors on cells,effectively prevent HaCat cells damaged by UV radiation,reduce the production of endothelin-1,inhibit melanin transfer,and regulate autophagy,which could provide some certain theoretical basis for ?-glucan from agaricus bisporus as ingredient of sunscreen and whitening skincare products. |
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