title: [Andrographolide inhibits extracellular signal-regulated kinase 1/2 signaling pathway in activated macrophages].
abstract: OBJECTIVE
  To investigate the effects of andrographolide on extracellular signal-regulated kinase 1/2 (ERK1/2) signaling pathway and tumor necrosis factor-α (TNF-α) expression in lipopolysaccharide (LPS)-activated macrophages.
  
  
  METHODS
  LPS-activated mouse peritoneal macrophages were cultured in media with different concentrations of andrographolide. Cytotoxicity of andrographolide was detected by cell counting kit-8. The macrophages were lysed, and then expressions of phosphorylated ERK1/2, JNK and p38 and nuclear factor-κB inhibitor (IκBα) protein were detected by Western blotting and TNF-α mRNA expression was detected by reverse transcription-polymerase chain reaction. Supernatants of the macrophages were used to detect content of TNF-α protein by enzyme-linked immunosorbent assay.
  
  
  RESULTS
  Andrographolide at 1-100 μg/mL showed no cytotoxicity on LPS-activated mouse peritoneal macrophages. Andrographolide inhibited ERK1/2 phosphorylation in LPS-activated murine peritoneal macrophages, which was concentration-dependent (P<0.01). Andrographolide at 1-25 μg/mL had no effects on phosphorylation levels of JNK and p38 and IκBα degradation in LPS-stimulated mouse peritoneal macrophages. In activated macrophages, TNF-α expression was inhibited by 12 μg/mL andrographolide and 20 μmol/L PD98059 (inhibitor of ERK1/2 signaling pathway) at both mRNA expression and protein secretion levels.
  
  
  CONCLUSIONS
  In LPS-activated macrophages, andrographolide may inhibit the expression of TNF-α by inhibiting ERK1/2 signaling pathway.
authors.affiliation: Department of Digestive Diseases, the 411 Hospital of People's Liberation Army, Shanghai 200081, China. ando_8171@yahoo.com.cn
authors.forename: Lin-hua
authors.initials: LH
authors.lastname: Qin
authors.forename: Jiao
authors.initials: J
authors.lastname: Lü
authors.forename: Lin
authors.initials: L
authors.lastname: Kong
authors.forename: Yun-xing
authors.initials: YX
authors.lastname: Shi
authors.forename: Yong-ping
authors.initials: YP
authors.lastname: Li
authors.forename: Guo-zhong
authors.initials: GZ
authors.lastname: Zhou
authors.forename: Zhi-wu
authors.initials: ZW
authors.lastname: Zheng
authors.forename: Lin
authors.initials: L
authors.lastname: Li
authors.forename: Xing-ying
authors.initials: XY
authors.lastname: Ji
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D004224:Diterpenes / Q000494:pharmacology*; D005260:Female; D020935:MAP Kinase Signaling System; D008262:Macrophage Activation; D008264:Macrophages / Q000187:drug effects* / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D010766:Phosphorylation; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D004224:Diterpenes; D014409:Tumor Necrosis Factor-alpha; C030419:andrographolide; D048052:Mitogen-Activated Protein Kinase 3