Abstract
Abstract The Jing-Fang powder n-butanol extract (JFNE) has anti-inflammatory properties; however, its active ingredient remains unknown. In addition, the mechanism by which JFNE exerts its anti-inflammatory effects on lipopolysaccharide (LPS) induced inflammation in RAW264.7 cells is yet to be explored. In this study, JFNE was isolated by chromatography to obtain fraction D. We found that pretreatment of LPS-induced RAW264.7 cells with JFNE and fraction D for 3 h significantly reduced the levels of nitric oxide (NO), interleukin-1beta (IL-1β), and tumor necrosis factor-alpha (TNF-α) in the supernatant of cell cultures, and fraction D could also reduce the level of interleukin-6 (IL-6). In addition,JFNE and fraction D significantly reduced the mRNA expression of inducible nitric oxide synthase (iNOS), IL-6, IL-1β, and TNF-α. JFNE and fraction D significantly inhibited the phosphorylation of proteins and mRNA expression levels of phosphoinositide 3-kinase (PI3K) and protein kinase B (PKB/AKT). Moreover, JFNE and fraction D significantly decreased the mRNA expression of iNOS, v-rel reticuloendotheliosis viral oncogene homolog A (RELA), and nuclear factor of kappa light polypeptide gene enhancer in B cells 1 (Nfκb1), while an increase in the mRNA expression of conserved helix-loop-helix ubiquitous kinase (CHUK) was observed. In addition, JFNE and fraction D down-regulated the protein expression of iNOS, nuclear factor-kappa B (NF-κB) (p50), and phosphorylated NF-κB (p65). These results show that JFNE and its isolated fraction D, exert specific anti-inflammation properties in LPS-stimulated RAW264.7 cells that are regulated by inhibition of the PI3K/AKT and NF-κB signaling pathways. Keywords: Jing-Fang powder, inflammation, RAW264.7, PI3K/AKT, NF-κB
SIGNIFICANCE STATEMENT none
- anti-inflammatory drugs
- cytokines
- drug efficacy
- endotoxins
- inositol triphosphate (IP3)
- natural products
- nfkb
- nitric oxide synthase
- pharmacodynamics
- pharmacogenetics
- The American Society for Pharmacology and Experimental Therapeutics