Effect Of SEB On The Expression Of S100A8 And S100A9 In HaCaT Cells

Objective:To detect the expression of S100A8、S100A9 proteins and extracellular cytokines such as Interleukin-la(IL-1α)、Interleukin-17(IL-17)、Tumor Necrosis Factor-a(TNF-a)、Interleukin-4(IL-4)staphylococcus aureus enterotoxin B(SEB)treated HaCaT cells.To further clarify the effect of damage associated molecular patterns(DAMP)molecules and innate immunity on the pathogenesis of AD.Methods:HaCaT cells from 3 to 5 generations were used for the experiments.MTT assay was used to determine the influence of cytoxicity in SEB stimulated HaCaT cells.The expression of DAMP molecules S100A8 and S100A9 proteins were detected by Western blot.The expression of IL-la,IL-17,TNF-α,IL-4 were measured by enzyme linked immunosorbent assay(ELISA).Results:1.MTT assay showed that there is no cytoxicity in HaCaT cells which were stimulated by different concentration of SEB at 24 hours(P>0.05).Similar results showed at 48 hours and 72 hours(P>0.05).2.Western blot analysis showed that compared with control group,the expression of intracellular S100A8 protein was significantly increased in 20 ng/ml SEB treated group;50 ng/ml SEB treated group and 100 ng/ml SEB treated group(P<0.01).The expression of extracellular S100A8 protein was detected by Western blot.The results showed that the expression of extracellular S100A8 protein was increased in 20 ng/ml SEB treated group compared with the control group(P<0.01).The expression of S100A9 protein was detected by Western blot.The expression of intracellular S100A9 protein was increased in the 20 ng/ml SEB group compared with the control group(P<0.01).The expression of extracellular S100A9 protein was also increased(P<0.01).3.The expression of IL-la was increased in HaCaT cells teated with different concentrations of SEB(P<0.01).The expression of IL-17 was significantly increased in 100 ng/ml SEB teated HaCaT cells,and the difference was statistically significant(P<0.01).TNF-α,IL-4 were not detected in HaCaT cells.Conclusion:1.SEB stimulates HaCaT cells to promote the expression of S100A8 and S100A9 proteins in AD pathogenesis,which indicates that changes in DAMP molecules S100A8 and S100 A9 proteins may affect the pathogenesis of AD.2.SEB stimulates HaCaT cells to promote the IL-la and IL-17 secretion,indicating that SEB may trigger symptoms of AD via increase of inflammatory cytokines IL-1α and IL-17.