Impacts of store-operated calcium entry on the function of epithelial barrier in atopic dermatitis
Objective:To explore the impacts of store-operated calcium entry(SOCE)on the epithelial barrier of atopic dermatitis(AD).Methods:AD model in 6-week BALB/c mouse was built by tape stripping and 1%DNCB challenging and mice were divided into 4 groups:control group,AD group,AD-BTP2 group,and AD-TCS group.The effects of inhibiting SOCE in vivo were evaluated by trans-epidermal water loss(TEWL)and histology of the epidermis.HaCaT cells were cultured in vi-tro and cell proliferation was tested by MTT assay after HDM and BTP2 stimulation.RT-PCR and Western-Blot of SOCE and epithelial barrier-related genes in HaCaT cells proceeded for expression changes and carried out FITC-dextran assay for cell barrier function evaluation.Results:TEWL and epidermis thickness significantly increased in AD group mice than in the control group(P<0.000 1).However,the BTP2-treated group showed a more significant increase in TEWL(P<0.05)and epi-dermis thickness(P<0.000 1)as compared with the AD group.HDM stimulation promotes HaCaT cell proliferation,but BTP2 showed no effects.The mRNA expression levels of SOCE-related genes had no significant change after HDM stimulation in HaCaT cells.The mRNA expression of SOCE-related proteins Orai1-Orai3,STIM1,STIM1,and SARAF in HaCaT cells after HDM stimulation did not differ from those in control cells,but the protein expression of Orai1 was significantly higher(P<0.05).Occludin and Loricrin,two barrier-related proteins,decreased after HDM stimulation,and BTP2 treatment could restore the expression of Occludin and elevate E-cadherin expression.HaCaT cells in the HDM group showed higher permeability of FITC-dextran than the control group,and BTP2(1 μmol/L)treatment significantly decreased the HDM-induced permeability changes.Conclu-sion:BTP2,the inhibitor of SOCE,shows opposite influences in the AD mouse model and HaCaT cell,which suggests the complexity of SOCE in type 2 inflammation,and further study of the underly-ing mechanisms needs to be conducted.
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