HMGB1-Binding Heptamer Confers Anti-Inflammatory Effects in Primary Microglia Culture.
- Author:
Il Doo KIM
1
;
Ja Kyeong LEE
Author Information
- Publication Type:Original Article
- Keywords: HMGB1; HBHP; inflammation; microglia
- MeSH: Brain; Culture Media; Cytokines; HMGB1 Protein; Infarction, Middle Cerebral Artery; Inflammation; Microglia*; Neuroprotective Agents
- From:Experimental Neurobiology 2013;22(4):301-307
- CountryRepublic of Korea
- Language:English
- Abstract: High mobility group box 1 (HMGB1) is an endogenous danger signal molecule. In the postischemic brain, HMGB1 is massively released during NMDA-induced acute damage and triggers inflammatory processes. In a previous study, we demonstrated that intranasally delivered HMGB1 binding heptamer peptide (HBHP; HMSKPVQ) affords robust neuroprotective effects in the ischemic brain after middle cerebral artery occlusion (MCAO, 60 minutes). In the present study, we investigated HBHP-induced anti-inflammatory effects on microglia activation. In LPS-treated primary microglia culture, HMGB1 was rapidly released and accumulated in culture media. Furthermore, LPS-conditioned media collected from primary microglia cultures (LCM) activated naive microglia and markedly induced NO and proinflammatory cytokines. However, the suppression of HMGB1 by siRNA-HMGB1, HMGB1 A box, or anti-HMGB1 antibody significantly attenuated LCM-induced microglial activation, suggesting that HMGB1 plays a critical role in this process. A pull-down assay using biotin-labeled HBHP showed that HBHP binds directly to HMGB1 (more specifically to HMGB1 A box) in LCM. In addition, HBHP consistently inhibited LCM-induced microglial activation and suppressed the inductions of iNOS and proinflammatory cytokines. Together these results suggest that HBHP confers anti-inflammatory effects in activated microglia cultures by forming a complex with HMGB1.
