AP-1 pathway-targeted inhibition of inflammatory responses in LPS-treated macrophages and EtOH/HCl-treated stomach by Archidendron clypearia methanol extract

Ethnopharmacological relevance: Archidendron clypearia Jack. (Fabaceae) is a representative ethnomedicinal herbal plant prescribed for various inflammatory diseases such as pharyngolaryngitis and tonsillitis. However, the pharmacology behind this plant's anti-inflammatory properties has not been fully understood. Therefore, in this study, the anti-inflammatory mechanism of a 95% methanol extract (Ac-ME) was explored. Materials and Methods: The anti-inflammatory mechanism of Ac-ME on the AP-1 activation pathway, which plays a critical role in the production of prostaglandin (PG)E₂ in RAW264.7 cells and peritoneal macrophages and in induction of acute gastritis caused by HCl/EtOH, was investigated using immunoblotting, immunoprecipitation analyses, and reporter gene activity assays. In particular, enzyme assays and HPLC analysis were employed to identify direct target enzymes of Ac-ME and to detect active chemical components from the plant extract. Results: Ac-ME clearly reduced the nuclear levels of total and phospho-forms of c-Jun, FRA-1, and ATF-2. Consequently, this extract suppressed both the production of PGE₂ in lipopolysaccharide (LPS)-activated RAW264.7 and peritoneal macrophage cells and PGE₂-dependent induction of gastritis lesion in stomach under EtOH/HCl exposure. Analysis of AP-1 upstream signalling revealed that the AP-1 activation pathway consisting of IRAK1, TRAF6, TAK1, MKK3/6, and p38 was predominantly inhibited by Ac-ME. Similarly, this extract directly blocked the enzyme activity of IRAK1, indicating that this enzyme is an inhibitory target of Ac-ME and is involved in the suppression of the AP-1 pathway. HPLC analysis showed that quercetin, which inhibits PGE₂ production, is an active component in Ac-ME. Conclusion: Ac-ME is an ethnomedicinal remedy with an IRAK1/p38/AP-1-targeted inhibitory property. Since AP-1 is a major inflammation-inducing transcription factor, the therapeutic potential of Ac-ME in other AP-1-mediated inflammatory symptoms will be further tested.