The Anti‐Inflammatory Role of Heme Oxygenase‐1 in Lipopolysaccharide and Cytokine‐Stimulated Inducible Nitric Oxide Synthase and Nitric Oxide Production in Human Periodontal Ligament Cells

Abstract

Although heme oxygenase-1 (HO-1) is involved in anti-inflammation, the mechanisms of its activity in regulating periodontal inflammation are largely unclear. Therefore, the aim of this study is to investigate the anti-inflammatory properties of HO-1 in lipopolysaccharide (LPS)- and proinflammatory cytokine-stimulated inducible nitric oxide synthase (iNOS) expression and nitric oxide (NO) production in human periodontal ligament (PDL) cells. PDL cells were treated with LPS plus a combination of tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta in serum-free media for 1 day. The production of NO was evaluated using a Griess reagent kit. The expression of iNOS and HO-1 proteins and mRNAs was evaluated using Western blotting and reverse transcriptase-polymerase chain reaction, respectively. Proinflammatory cytokines and LPS triggered iNOS and HO-1 expression and NO production in PDL cells. HO-1 inhibitor and HO-1 small interfering RNA (siRNA) attenuated the LPS- and cytokine-stimulated NO release and iNOS and HO-1 expression. Specific inhibitors of p38, extracellular signal-regulated kinase (ERK), and c-Jun N-terminal kinase (JNK) mitogen-activated protein kinases phosphatidylinositol 3-kinase (PI3K), nuclear factor-kappa B (NF-kappaB), and protein kinase C delta (PKC-delta) greatly reduced the levels of iNOS and HO-1 expression induced by LPS plus cytokines. Collectively, these data suggested that HO-1 inhibition blocked LPS- and proinflammatory cytokine-stimulated iNOS expression and NO production in PDL cells via a mechanism that involves p38, ERK, PI3K, NF-kappaB, and PKC-delta. Thus, the regulation of HO-1 activity may be a therapeutic strategy for periodontal disease.