The Effect Of Genistein In The UA-induced Damage In HaCat Cells

Objective:Establishig a photoaging model in vitro experiments to investigate the relationship between the dose of UVA irradiation and the degree of Hacat cell injury,to explore the changes of cellular senescence-associated proteins during the injury process,to explore the protective effect of genistein on Hacat cell injury,and to explore the effect of genistein on transcription factor Bach1 in the cellular UVA-related injury model.Methods:(1)0 J/cm~2,3.6 J/cm~2,7.2 J/cm~2,10.8 J/cm~2,14.4 J/cm~2dose of UVA was irradiated to Ha Cat cells respectively,and cells were observed 24h after irradiation,then testing the proliferation activity of the cells by CCK8 technology.(2)Using 0 umol/L、0.01umol/L、0.1 umol/L、1 umol/L、10 umol/L、20 umol/L、40 umol/L、60 umol/L、80 umol/L、100 umol/L genistein to culture Ha Cat cells respectively,and the proliferation activity of the cells was tested by CCK8 technology 48h after.This study set the concentration of genistein at 20μM for the following experiments.(3)There were three groups of Ha Cat cells.Group A was a control group.Group B was a UVA irradiation with genistein group.Groups C was a UVA irradiation group.Western blot was used to determine the expression of the Bach1 protein and aging-related protein p53 in group A,B and C.Analyze the results.Results:(1)After UVA irradiation at different doses,the cells in each group showed damage manifestation,and cell activity was reduced compared to the control group.(2)Cells treated with media of different concentrations of genistein,low concentration of dynein had little effect on cell growth,and high concentration of genistein treatment inhibited cell proliferation.(3)The expression of senescence-related protein p53 increased in group C compared with group A,and group B was lower than group C.The expression of transcription factor Bach1 increased in group C compared with group A,and group B was lower than group C.Conclusion:Long-wave UV irradiation can cause a decrease in the activity of human cells,and the stronger the irradiation dose,the more severe the damage to cell activity;7.2 J/cm~2 dose of UVA irradiation can cause cell damage,cell morphological changes,and increased expression of senescence protein p53 and transcription factor Bach1 in damaged cells;high concentration of dynein can reduce the proliferation activity of human immortalized keratinocytes in culture;A certain concentration of genistein(20umol/L)followed by 7.2 J/cm~2 long-wave UV irradiation could reduce the damage caused by UV irradiation,a specific concentration of genistein could play a protective role in enhancing the resistance of human immortalized keratinocytes to UV irradiation.