Anti-inflammatory effects of flavonoids and phenylethanoid glycosides from Hosta plantaginea flowers in LPS-stimulated RAW 264.7 macrophages through inhibition of the NE-κB signaling pathway

Background: The flower of Hosta plantaginea (Lam.) Aschers has traditionally been used in China as an important Mongolian medicine for the treatment of inflammatory diseases with limited scientific evidence. In previous studies, 16 flavonoids and 3 phenylethanoid glycosides (1-19) were isolated from the ethanolic extract of H. plantaginea flowers. Nevertheless, the anti-inflammatory effects of these constituents remain unclear. In the present study, the anti-inflammatory effects of these 19 constituents and their underlying mechanisms were assessed in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. Methods: The viability of RAW 264.7 macrophages was detected by Cell Counting Kit-8 (CCK-8) assay. Meanwhile, nitric oxide (NO) production was measured by Griess assay, while the secretion of tumor necrosis factor alpha (TNF-alpha), prostaglandin E2 (PGE2), interleukin 1 beta (IL-1 beta) and IL-6 in LPS-induced macrophages was determined by enzyme-linked immunosorbent assay (ELISA). Furthermore, the protein expression of nuclear factor kappa B (NF-kappa B) p65 and phosphorylated NF-kappa B p65 was evaluated by Western blot analysis. Results: All constituents effectively suppressed excessive NO production at a concentration of 40 pM with no toxicity to LPS-induced RAW 264.7 macrophages. Among them, five flavonoids (1, 4-6 and 15) and one phenylethanoid glycoside (17) remarkably prevented the overproduction of NO with median inhibitory concentration (IC50) values in the range of 12.20-19.91 mu M. Moreover, compounds 1, 4-6, 15 and 17 potently inhibited the secretion of TNF-alpha, PGE2, IL-1 beta and IL-6, and had a prominent inhibitory effect on the down-regulation of the phosphorylated protein level of NF-kappa B p65. Conclusion: Taken together, compounds 1, 4-6, 15 and 17 may be useful in managing inflammatory diseases by blocking the NF-kappa B signaling pathway and suppressing the overproduction of inflammatory mediators.