HO-1 Induction by CO-RM2 Attenuates TNF-α-Induced Cytosolic Phospholipase A2 Expression via Inhibition of PKCα-Dependent NADPH Oxidase/ROS and NF-κB

Rheumatoid arthritis (RA) is characterized by chronic inflammatory infiltration of the synovium and elevation of proinflammatory cytokines. Cytosolic phospholipase A(2) (cPLA(2)) is involved in the development of inflammatory diseases. Heme oxygenase-1 (HO-1) has been shown to possess anti-inflammatory properties. The objective of the study was to investigate the detailed mechanisms of TNF-alpha-induced cPLA(2) expression and to determine whether carbon monoxide releasing molecule-2 (CO-RM2) suppresses TNF-alpha-induced expression of NF-kappa B-related proinflammatory genes, including cPLA(2), via HO-1 induction in RA synovial fibroblasts (RASFs). Here, we reported that TNF-alpha-induced cPLA(2) expression was mediated through TNFR1/PKC alpha-dependent signaling pathways, including NADPH oxidase (NOX) activation/ROS production and NF-kappa B activation. CO-RM2 significantly suppressed TNF-alpha-induced cPLA(2) expression by inhibiting the ROS generation and the phosphorylation of NF-kappa B p65 and IKK alpha/beta, but not the phosphorylation of p38 MAPK and JNK1/2. These results were further confirmed by a ChIP assay to detect the NF-kappa B DNA-binding activity. Our results demonstrated that induction of HO-1 by CO-RM2 exerted anti-inflammatory and antioxidant effects which were required in concert to prevent the activation of NF-kappa B leading to induction of various inflammatory genes implicated in the pathogenesis of RA.