Anti-inflammatory Activity And Mechanism Of An Artemisia Vulgaris L.sesquiterpene Lactone Artemvulactone E In Vivo And In Vitro

Objective:In this paper,Artemvulactone E（AE）,a new compound isolated and extracted from Artemisia vulgaris L.in the laboratory,was used as the research object to study its anti-inflammatory activity in vitro and in vivo,and to explore its anti-inflammatory mechanism,so as to lay a theoretical foundation for the development of new anti-inflammatory drugs.Methods:MTT assay was used to detect cytotoxicity.RT-q PCR,Griess reagent method,ELISA and Western blot were used to explore the inhibitory effect of AE on proinflammatory cytokines from gene and protein levels.The anti-inflammatory mechanism of AE was studied by Western blot and nuclear cytoplasmic separation experiment.Finally,the zebrafish inflammation model induced by microinjection of LPS into yolk sac was established to explore the in vivo anti-inflammatory activity of AE,and the in vivo anti-inflammatory activity of AE was evaluated by survival rate,heart rate,histopathology and the recruitment of neutrophils and macrophages at the inflammatory site.Results:The IC₅₀of RAW264.7 cells treated with AE for 12 h and 24 h were10.19 and 6.41μM,respectively.AE can inhibit the expression of TNF-α,IL-6,i NOS and COX-2 from the m RNA level and protein level,inhibit the extracellular secretion of TNF-α,IL-6 and NO,and reduce the production of intracellular NO and ROS,showed good anti-inflammatory activity in vitro.In terms of mechanism,AE can inhibit the activation of NF-κB,MAPK and IL-6/JAK/STAT3 signaling pathways.In vivo studies,AE can reduce the mortality and heart rate rise of zebrafish embryos caused by LPS,reduce the infiltration of inflammatory cells in the yolk sac,and inhibit the recruitment of neutrophils and macrophages to the yolk sac.Conclusions:AE has good anti-inflammatory activity,and it can reduce the expression and secretion of pro-inflammatory cytokine TNF-αand IL-6,down regulate the m RNA and protein expression of inflammatory mediators i NOS and COX-2,and reduce the production of intracellular NO and ROS.In addition,AE can improve the survival rate of infected zebrafish and attenuate LPS-induced inflammatory response in zebrafish embryos.Mechanistically,AE synergistically exert anti-inflammatory effects through NF-κB signaling pathway,MAPK signaling pathway and IL-6/JAK/STAT3 signaling pathway.