Journal: British Journal of Pharmacology

Article Title: Up‐regulation of PYK2/PKCα‐dependent haem oxygenase‐1 by CO‐releasing molecule‐2 attenuates TNF‐α‐induced lung inflammation

doi: 10.1111/bph.14094

Figure Lengend Snippet: CORM‐2 induces HO‐1 expression via AP‐1. (A) HPAEpiCs were pretreated with Tanshinone IIA (TSA IIA; 0.01, 0.1 and 1 μM) for 1 h and then incubated with 50 μM CORM‐2 for 16 h. The levels of HO‐1 protein were determined. (B) Cells were pretreated with Tanshinone IIA (1 μM) for 1 h and then incubated with 50 μM CORM‐2 for 6 h. The HO‐1 mRNA levels were determined by real‐time PCR. (C, D) Cells were pretreated without or with PF431396 (3 μM), Gö6976 (10 μM), Gö6983 (10 μM) or U0126 (10 μM) for 1 h and then incubated with 50 μM CORM‐2 for 2 h. The c‐Fos mRNA levels were measured by real‐time PCR. (E) Cells were transfected with scrambled or c‐Fos siRNA and then incubated with 50 μM CORM‐2 for 16 h. The levels of c‐Fos, HO‐1 and β‐actin protein were determined. Data are expressed as mean ± SEM of five independent experiments. *P < 0.05, as compared with the cells exposed to CORM‐2 alone (A, B, D) or vehicle (0.5% DMSO) alone (C). (F) Schematic signalling pathways are involved in CORM‐2‐induced HO‐1 expression. CORM‐2 activates the PYK2/PKCα pathway to enhance ERK1/2 (p44/42 MAPK) phosphorylation, which in turn initiates the activation of AP‐1 and ultimately induces HO‐1 expression in HPAEpiCs. Moreover, pretreatment with CORM‐2 inhibits TNF‐α‐induced lung inflammation via the induction of HO‐1.

Article Snippet: PF431396 (Han et al., 2009a , b ), Ro‐318220 , Gö6976 (Martiny‐Baron et al., 1993 ), Gö6983 (Stempka et al., 1999 ), U0126 (Favata et al., 1998 ) and Tanshinone IIA (Jang et al., 2003 ) were from Biomol (Plymouth Meeting, PA, USA).

Techniques: Expressing, Incubation, Real-time Polymerase Chain Reaction, Transfection, Phospho-proteomics, Activation Assay