Articles

Article

Original Article

Exp Neurobiol 2013; 22(4): 301-307

Published online December 30, 2013

https://doi.org/10.5607/en.2013.22.4.301

© The Korean Society for Brain and Neural Sciences

HMGB1-Binding Heptamer Confers Anti-Inflammatory Effects in Primary Microglia Culture

Il-Doo Kim and Ja-Kyeong Lee*

Department of Anatomy, Inha University School of Medicine, Incheon 400-712, Korea

Correspondence to: *To whom correspondence should be addressed.
TEL: 82-32-890-0913, FAX: 82-32-884-2105
e-mail: jklee@inha.ac.kr

Received: December 6, 2013; Revised: December 17, 2013; Accepted: December 17, 2013

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 naïve 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.

Keywords: HMGB1, HBHP, inflammation, microglia