Objective:In order to elucidate the biological activity of the co-cultured adventitious roots (ARs) of Echinacea pallida and Echinacea purpurea and provide theoretical basis for its application,and the anti-inflammatory activities and potential mechanisms of co-cultured ARs were studied.Methods:The experimental materials were obtained by bioreactor co-culture technology and used in the activity research.In this study,mouse macrophages induced by lipopolysaccharide (LPS) were used as in vitro model.Different concentrations of AR extract (50-400 g/mL) were used to treat cells.The expres-sion of pro-inflammatory cytokines was determined using enzyme linked immunosorbent assay.The inducible nitric oxide synthase and cyclooxygenase-2 expression,mitogen-activated protein kinase(MAPK) phosphorylation,and the inhibitor of nuclear factor-kappa B-α levels were determined by the Western blot analysis.Results:In the co-cultured ARs,total flavonoids and total caffeic acid were determined,and the contents of both bioactive compounds were significantly higher than those ARs from the single-species culture.Compared with the control group,the large amount of pro-inflammatory mediators was released after LPS stimulation.However,in the extract groups with different concentrations (25,50,and 100 g/mL),the production of these pro-inflammatory mediators was inhibited in a dose-dependent manner.Furthermore,the levels of phosphorylation of MAPK proteins,including p-p38,p-c-Jun N-terminal kinase,and p-extracellular regulated protein kinases were significantly (P ＜ 0.05) decreased in the extract groups,revealing that the AR extract probably involved in regulating the MAPK signaling pathway.Conclusion:Collectively,our findings suggested that the co-cultured ARs of E.pallida and E.purpurea can inhibit production of pro-inflammatory mediators in mouse peritoneal macrophages and possess the anti-inflammatory effect by regulating MAPK signaling pathways.