Immunomodulatory effect of Glossogyne tenuifolia in murine peritoneal macrophages and splenocytes

Glossogyne tenuifolia Cass., a medicinal plant native to Taiwan, is traditionally used as an anti-inflammatory remedy. Oleanolic acid and luteolin-7-glucoside have been previously identified as active components of Glossogyne tenuifolia in the murine macrophage-like cell line, RAW264.7. Current study investigates the effect and mechanism of the ethanol extract of Glossogyne tenuifolia (GT) and its major constituents on the release of inflammatory mediators in activated elicited murine peritoneal macrophages and splenocytes. Our results showed that GT (up to 0.15 mg/ml) inhibited the production of proinflammatory mediators, TNF-alpha, IL-1beta, IL-6, nitric oxide (NO) and prostaglandin E(2) (PGE(2)) in LPS-activated macrophages, and IFN-gamma in PHA-activated splenocytes. GT also inhibited LPS-activated murine iNOS and COX-2 promoter activities in transiently transfected RAW264.7 cells. The major constituents, oleanolic acid and luteolin-7-glucoside, as well as its aglycone, luteolin, inhibited the release of NO, PGE(2), TNF-alpha and IL-1beta in activated peritoneal macrophages. However, only luteolin-7-glucoside and luteolin were able to reduce IFN-gamma release in PHA-stimulated splenocytes. To further investigate the possible mechanisms that interfere with LPS- and PHA-signaling, this study focused on nuclear factor-kappaB activation signaling pathways. Our results demonstrate that GT (0.075-0.15 mg/ml) treatment reduces nuclear factor-kappaB (NF-kappaB) DNA binding activity, as demonstrated by electrophoretic mobility shift assay (EMSA). Collectively, the results suggest that GT inhibits proinflammatory mediator synthesis in activated murine peritoneal macrophages and splenocytes, in part through NF-kappaB-dependent pathways.