Inhibition of LPS-induced expression of iNOS and COX-2 on extracts of Acanthopanax leucorrhizus (Oliv.) Harms stems

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Abstract

Abstract Acanthopanax leucorrhizus is an endemic medicinal plant growing abundantly in the northwest region of China. The roots and stem barks have been widely used to treat rheumatism, numbness, contracture, quadriplegia, hemiplegia, traumatic injury, edema, and itchy skin, although the anti-inflammatory effects of the extracts of A. leucorrhizus have not been assessed until now. To investigate the anti-inflammatory effects on macrophages of various extracts from A. leucorrhizus stems, alterations in the inducible nitric oxide synthase (iNOS)-mediated cyclooxygenase-2 (COX-2) expression and inflammatory cytokine production were measured in lipopolysaccharide (LPS)-activated RAW264.7 cells. Briefly, the dichloromethane extract prepared from stems of A. leucorrhizus (ALSDC) effectively inhibited NO production in LPS-stimulated cells and significantly reduced the production of pro-inflammatory cytokines IL-6 and TNF-α at a dose of 40 µg/mL. We also confirmed a dose-dependent and significant inhibition of iNOS and COX-2 protein expression. In conclusion, ALSDC exerted strong inhibitory effect on the expression of iNOS and COX-2 protein in LPS-induced RAW 264.7 macrophages and these results provide strong evidence to suggest that ALSDC may be considered as an important candidate for the treatment of inflammatory-related diseases in the future.

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Red Ginseng Marc Oil Inhibits iNOS and COX-2 via NFκB and p38 Pathways in LPS-Stimulated RAW 264.7 Macrophages

Min-Ji Bak, Soon-Gi Hong, Jong-Won Lee … (2012)

In this study, we investigated the anti-inflammatory effects of red ginseng marc oil (RMO) in the RAW 264.7 macrophage cell line. RMO was prepared by a supercritical CO2 extraction of waste product generated after hot water extraction of red ginseng. RMO significantly inhibited the production of oxidative stress molecules such as nitric oxide and reactive oxygen species in lipopolysaccharide (LPS)-activated RAW 264.7 cells. Levels of inflammatory targets including prostaglandin E2, tumor necrosis factor-α, interleukin (IL)-1β and IL-6 were also reduced after the treatment with RMO. In addition, RMO diminished the expressions of inducible nitric oxide synthase and cyclooxygenase 2 at both mRNA and protein levels. Blockade of nuclear translocation of the p65 subunit of nuclear factor κB (NFκB) was also observed after the treatment of RMO. Furthermore, RMO decreased the phosphorylations of p38 mitogen-activated protein kinase (MAPK) and its upstream kinases including MAPK kinases 3/6 (MKK3/6) and TAK 1 (TGF-β activated kinase 1). Gas chromatographic analysis on RMO revealed that RMO contained about 10% phytosterols including sitosterol, stigmasterol and campesterol which may contribute to the anti-inflammatory properties of RMO. Taken together, these results suggest that the anti-inflammatory effect of RMO in LPS-induced RAW 264.7 macrophages could be associated with the inhibition of NFκB transcriptional activity, possibly via blocking the p38 MAPK pathway.