title: Selective alpha7-nicotinic acetylcholine receptor agonist GTS-21 improves survival in murine endotoxemia and severe sepsis.
abstract: OBJECTIVE
  Tumor necrosis factor and high mobility group box 1 are critical cytokine mediators of inflammation. The efferent vagus nerve inhibits cytokine release through alpha7-nicotinic acetylcholine receptor-mediated cholinergic signaling. Here we studied whether GTS-21, a selective alpha7-nicotinic acetylcholine receptor agonist, inhibits proinflammatory cytokines in vitro and in vivo and improves survival in murine endotoxemia and severe sepsis.
  
  
  METHODS
  Randomized and controlled in vitro and in vivo study.
  
  
  METHODS
  Research laboratory and animal facility rooms.
  
  
  METHODS
  RAW 264.7 cells and BALB/c mice treated with endotoxin or subjected to cecal ligation and puncture (CLP).
  
  
  METHODS
  RAW 264.7 cells were exposed to endotoxin (4 ng/mL or 10 ng/mL) in the presence or absence of GTS-21 (1-100 muM), and tumor necrosis factor and high mobility group box 1 release and nuclear factor-kappaB activation were analyzed. Mice were treated with GTS-21 (0.4 mg/kg or 4 mg/kg, intraperitoneally) or saline 30 mins before endotoxin (6 mg/kg, intraperitoneally), and serum tumor necrosis factor was analyzed 1.5 hrs after the onset of endotoxemia. In survival experiments, mice were treated with GTS-21 (0.4 or 4.0 mg/kg, intraperitoneally) or saline 30 mins before and 6 hrs after endotoxin and then twice daily for 3 days. Severe sepsis was induced by CLP. Mice were treated with GTS-21 (4 mg/kg) or saline immediately and 6 hrs and 24 hrs after CLP, and serum high mobility group box 1 was analyzed 30 hrs after CLP. In survival experiments, GTS-21 (0.4 or 4 mg/kg) treatment was initiated 24 hrs after CLP and continued twice daily for 3 days.
  
  
  RESULTS
  GTS-21 dose-dependently inhibited tumor necrosis factor and high mobility group box 1 release and nuclear factor-kappaB activation in vitro. GTS-21 (4 mg/kg) significantly inhibited serum tumor necrosis factor during endotoxemia and improved survival (p < .0001). GTS-21 (4 mg/kg) significantly inhibited serum high mobility group box 1 levels in CLP mice and improved survival (p < .0006).
  
  
  CONCLUSIONS
  These findings are of interest for the development of alpha7-nicotinic acetylcholine receptor agonists as a new class of anti-inflammatory therapeutics.
authors.affiliation: Laboratory of Biomedical Science, The Feinstein Institute for Medical Research, Manhasset, NY, USA. vpavlov@nshs.edu
authors.forename: Valentin A
authors.initials: VA
authors.lastname: Pavlov
authors.forename: Mahendar
authors.initials: M
authors.lastname: Ochani
authors.forename: Li-Hong
authors.initials: LH
authors.lastname: Yang
authors.forename: Margot
authors.initials: M
authors.lastname: Gallowitsch-Puerta
authors.forename: Kanta
authors.initials: K
authors.lastname: Ochani
authors.forename: Xinchun
authors.initials: X
authors.lastname: Lin
authors.forename: Jelena
authors.initials: J
authors.lastname: Levi
authors.forename: William R
authors.initials: WR
authors.lastname: Parrish
authors.forename: Mauricio
authors.initials: M
authors.lastname: Rosas-Ballina
authors.forename: Christopher J
authors.initials: CJ
authors.lastname: Czura
authors.forename: Gregory J
authors.initials: GJ
authors.lastname: Larosa
authors.forename: Edmund J
authors.initials: EJ
authors.lastname: Miller
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Tracey
authors.forename: Yousef
authors.initials: Y
authors.lastname: Al-Abed
mesh_terms: D000818:Animals; D001597:Benzylidene Compounds / Q000494:pharmacology*; D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D019446:Endotoxemia / Q000188:drug therapy* / Q000401:mortality; D024243:HMGB1 Protein / Q000037:antagonists & inhibitors; D066298:In Vitro Techniques; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D016328:NF-kappa B / Q000037:antagonists & inhibitors; D018722:Nicotinic Agonists / Q000494:pharmacology*; D011725:Pyridines / Q000494:pharmacology*; D011978:Receptors, Nicotinic / Q000378:metabolism*; D018805:Sepsis / Q000188:drug therapy* / Q000401:mortality; D012720:Severity of Illness Index; D015996:Survival Rate; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors; D064569:alpha7 Nicotinic Acetylcholine Receptor
chemical_list: D001597:Benzylidene Compounds; C579166:Chrna7 protein, mouse; D024243:HMGB1 Protein; D016328:NF-kappa B; D018722:Nicotinic Agonists; D011725:Pyridines; D011978:Receptors, Nicotinic; D014409:Tumor Necrosis Factor-alpha; D064569:alpha7 Nicotinic Acetylcholine Receptor; C088936:3-(2,4-dimethoxybenzylidene)anabaseine