The Effect Of Artemether On The Expression Of Fas And Bcl-2 In HaCaT Cells After Exposure To UV

OBJECTIVE In order to approach the mechanism of artemether treated photodermatoses, to observe the effect of treated with different concentrations of artemether on the expression of Fas and Bcl-2 in HaCaT cells after exposure to UV which is invariably dosage and different wavelength.METHODS Subconfluent HaCaT cells were shammed or irradiated with 4．00J/cm~2 of UVA or 45．00mJ/cm~2 of UVB respectively. The cytoactive and the level of Fas and Bcl-2 protein in HaCaT cells, which were irradiated by Ultraviolet and treated with hydroxychloroquine and different concentrations of artemether ,were detected by MTT method,immunohistochemistry and flow cytometry. Group as follow: Subconfluent HaCaT cell was divided into no UV irradiation groups,UVA irradiation groups,UVA+Q groups (dishes were added 50μg/mL hydroxychloroquine of final concentration after UVA irradiation ),UVA+H1 groups,UVA+H2 groups,UVA+H3 groups,UVA+H4 groups (dishes were added 25μg/mL,50μg/mL,100μg/mL,150μg/mL artemether of final concentration after UVA irradiation),UVB groups,UVB+Q groups,UVB+H1 groups,UVB+H2 groups,UVB+H3 groups,UVB+H4 groups(dishes were added corresponding final concentration of drug after UVB irradiation).RESULTS MTT results: The cytoactive didn' t change obviously in HaCaT cells which were treated with hydroxychloroquine and low dosage (≤50μg/ml) of artemether, but above 100μg/ml of artemether can increase. When concentrations exceeded 200μg/ml, a large number of cells suspended on culture solution.Flow cytometry results: 1．Compared to cells without UV irradiation, the expression of Fas was upregulated in HaCaT cells after UVA and UVB irradiation. 2．Compared to cells with UV irradiation, UVA+Qgroups,UVB+H1groups and UVA+H2groups downregulated the expression of Fas. 3．Compared to cells without UV irradiation, the expression of Bcl-2 was downregulated in HaCaT cells after UVA and UVB irradiation. 4．Compared to cells with UV irradiation, UVA+H1 groups and UVB+H3 groups upregulated the expression of Bcl-2 in HaCaT cells.Immunohistochemistry results: 1．Compared to cells without UV irradiation, the expression of Fas was upregulated in HaCaT cells after UVA and UVB irradiation. 2．Compared to cells with UV irradiation, UVA+Q groups,UVA+H1 groups,UVA+H2 groups and UVA+H3 groups downregulated the expression of Fas, UVB+Q groups,UVB+H1 groups,UVB+H2 groups and UVB+H3 groups changed uniformly. 3．Compared to cells without UV irradiation, the expression of Bcl-2 was downregulated in HaCaT cells after UVA and UVB irradiation. 4．Compared to cells with UV irradiation, UVA+Q groups,UVB+Q groups,UVB+H1 groups,UVB+H2 groups and UVB+H3 groups upregulated the expression of Bcl-2 in HaCaT cells.CONCLUSION Low dosage of artemether could prevent apoptosis of HaCaT cells , it has protective effect on HaCaT cells damaged from UV irradiation. Artemether might affect the expression of Fas and Bcl-2 protein, then reduce UV-induced apoptosis of HaCaT cells.