Signaling molecules involved in production and regulation of IL-1beta by murine peritoneal macrophages in vitro on treatment with Concanavalin A.

摘要

In the present study we report the activation of murine peritoneal macrophages in vitro on treatment with Concanavalin A (ConA). ConA (10 microg/ml) treatment of macrophages resulted in the transcription of IL-1beta gene at 16 h and maximum production of IL-1beta at 24 h. To investigate the signaling molecules involved in the production of IL-1beta different pharmacological inhibitors were used. It was observed that genestein, wortmannin, H-7, TMB-8, PD98059, SB202190, and tyrophostin (AG490) down regulated the expression of IL-1beta. These observations suggested the involvement of tyrosine kinase, PI3 kinase, protein kinase C, p42/44, p38, Ca(++) and JAK2 signaling molecules in ConA induced production of IL-1beta by macrophages. Maximum protein tyrosine kinase activity and expression of PI3K in macrophages was seen at 5 min, PKC activity and Ca(++) release was found at 10 min after ConA treatment. Maximum expression of phospho-JAK2 at 2.5-5 min, phospho-p42/44 at 5-60 min, phospho-p38 at 15-30 min, phospho-IkappaB and phospho-Stat1 at 30-60 min and phospho-ELK1, c-Fos, phospho-Stat3 at 60 min of ConA treatment was observed. Pharmacological inhibitors were also used to check the cascade of activation of tyrosine kinase, PKC, PI3 kinase, p42/44, p38, JAK kinase and release of Ca(++) from intracellular storage to sort out the signaling pathways involved in the release of IL-1beta by macrophages on treatment with ConA in vitro.