Inducing effect of IL-1βabout the release of HMGB1 in human bronchial epithelial cells / 重庆医学

ABSTRACT

Objective To investigate the effect of IL‐1βon the expression and release of high mobility group box 1 (HMGB1) from normal human bronchiolar epithelial cell (HBE) .Methods HBE135‐E6E7 was developed and methyltetrazolium (MTT) as‐say was used to assess the viability of HBE cell line under different concentration of IL‐1β.The mRNA expression of HMGB1 in HBE after stimulating with IL‐1βwere determined by Real‐time PCR;the level and location of HMGB1 in the cytoplasm ,nucleus and culture medium of HBE after stimulating with IL‐1β were detected by Western blot and ELISA assay .The expression and translocation of HMGB1 of HBE after stimulating with IL‐1β were detected by Immunofluorescence .Results 0 .1 ,1 .0 ,10 .0 ng/mL IL‐1βdid not influence the cell viability of HBE ;IL‐1β increased the mRNA expression of HMGB1 in HBE in does‐and time‐dependent manner and increased the protein expression of HMGB1 in HBE .In comparison with control group ,the levels of HMGB1 in the culture medium significantly increased after stimulation with IL‐1βat 1 .0 ,10 .0 ng/mL for 24 h(P<0 .05)in the dose dependent experiments;and in time dependent experiments ,10 .0 ng/mL IL‐1βsignificantly increased HMGB1 level in the cul‐ture medium after stimulation for both 12 h and 24 h (P< 0 .05) .After stimulation with IL‐1β (10 .0 ng/mL ) for 24 h ,the HMGB1 expression in cytoplasm significantly increased in plasmosin and decreased in nucleus .HMGB1 translocated from nuclei to cytoplasm after stimulation with IL‐1β(10 .0 ng/mL ) for 24 h .Conclusion IL‐1βcould induce HMGB1 expression and release in human bronchial epithelial cells ,which indicates that HMGB1 may involve in IL‐1β‐mediated chronic airway inflammation .