The Role Of Caveolae And Caveolin-1 On IL-1?/IFN-?/MIP-2 Production In Alveolar Macrophages

Objective:To investigate the possible effection of caveolae structure and cav-1 expression on IL-1?,IFN-y,MIP-2 production in alveolar macrophages and to discover the role of CSD peptide(cav-1 scaffolding domain peptide)on IL-1?expression in alveolar macrophages.Methods:Alveolar macrophages(AM)were isolated from bronchoalveolar lavage fluid.Immunofluorescence Microscopy and Western blot were used to identify cav-1 expression in Balb/c mice AM.The concentration of IL-1?,IFN-? and MIP-2 in culture medium of RAW264.7 cells treated with Filipin?(2.5?g/ml)for 30min were measured by ELISA.The amount of IL-1? mRNA was measured by Real-Time PCR(RT PCR).The levels of IL-1?,IFN-? and MIP-2 were measured by ELISA.Proteins of RAW264.7 cells treated with LPS(1?g/ml)for 15 min,30 min,1 h,2 h,4 h,6 h were isolated to analyze IL-1? by western blot.The possible effection of CSD peptide in different dose and treatment before or after LPS administration)on IL-1?expression were detected by Western blot.Results:1.Cav-1 is expressed in Balb/c mice AM.2.Filipin? treatment upregulated the production of pro-inflammatory cytokines IL-1?,IFN-? and chemotactic factor MIP-2 in RAW264.7 cells(P<0.05).3.LPS-challenge decreased cav-1 and increased IL-1?,IFN-y and MIP-2(P<0.05).The present of Filipin? enhanced upregulation of these three cytokines induced by LPS(P<0.05).4.CSD peptide pretreatment for 6 hours attenuated the IL-1? production induced by LPS,while 30-hour-treatment with CSD peptide increased the IL-1? production induced by LPS(P<0.05).Conclusions:1.Caveolae and cav-1 help to regulate inflammation development.2.The caveolae structure and cav-1 expression could modulate the production of IL-1?,IFN-? and MIP-2 in macrophages.3.CSD peptide treatment in right time could attenuate the IL-1? production induced by LPS.