title: Effects of sevoflurane postconditioning on cell death, inflammation and TLR expression in human endothelial cells exposed to LPS.
abstract: BACKGROUND
  Sevoflurane is an anesthetic agent which also participates in protective mechanisms in sepsis, likely due to anti-inflammatory properties. A key tissue in sepsis is the endothelium, which expresses TLR2 and TLR4 receptors, known regulators of inflammatory mechanisms and potential therapeutic targets for this pathology. In this context, we explored the effect of sevoflurane postconditioning in an in vitro sepsis model.
  
  
  METHODS
  Primary cultures of human umbilical vein endothelial cells were used for two different experiments. In the first set, cultures were placed in an airtight incubation chamber and exposed to different concentrations of sevoflurane (0,1,3 or 7% vol,) for 1 hour. In the second set, lipopolysaccharide from Escherichia coli 0111:B4 (1 μg/mL) was added to culture medium for 3 hours and cells were subsequently exposed to sevoflurane (0,1,3 or 7% vol,) for 1 hour as explained before. In both cases, cell viability was measured by MTT and Trypan blue assays, TLR2 and TLR4 expression were analyzed by flow cytometry, and TNFα and IL-6 levels were quantified in cell culture media by an immunoassay immediately after exposure, at 6 and 24 hours.
  
  
  RESULTS
  Exposure to 3% sevoflurane decreased TLR2 at 24 hours and TLR4 at 6 and 24 hours (both p<0.05), whereas exposure to 7% decreased TLR4 expression at 6 hours (p<0.05). Both 3 and 7% sevoflurane decreased TNF-α and IL-6 levels at 24 hours (both p<0.05). In LPS-stimulated cultures, exposure to 3% sevoflurane was cytoprotective at 6 and 24 hours (p<0.05) compared with control, and decreased TLR2 and TLR4 expression at 24 hours (p<0.05); whereas 7% decreased TLR4 expression at 24 hours (p<0.05). Both 3% and 7% sevoflurane decreased TNF-α and IL-6 levels at 24 hours (both p<0.05).
  
  
  CONCLUSIONS
  Postconditioning with the halogenated anesthetic agent sevoflurane after LPS stimulation shows a cytoprotective effect in an in vitro model, decreasing cell death and reducing TLR2 and TLR4 expression as well as levels of the inflammatory mediators TNF-α and IL-6 in human endothelial cells.
authors.affiliation: Critical Patient Translational Research Group, Department of Anesthesiology, Intensive Care and Pain Management, Hospital Clínico Universitario, IDIS, University of Santiago de Compostela, Santiago de Compostela, Spain.
authors.forename: Raquel
authors.initials: R
authors.lastname: Rodríguez-González
authors.forename: Aurora
authors.initials: A
authors.lastname: Baluja
authors.forename: Sonia
authors.initials: S
authors.lastname: Veiras Del Río
authors.forename: Alfonso
authors.initials: A
authors.lastname: Rodríguez
authors.forename: Jaime
authors.initials: J
authors.lastname: Rodríguez
authors.forename: Manuel
authors.initials: M
authors.lastname: Taboada
authors.forename: David
authors.initials: D
authors.lastname: Brea
authors.forename: Julián
authors.initials: J
authors.lastname: Álvarez
mesh_terms: D016923:Cell Death; D002470:Cell Survival; D003470:Culture Media / Q000494:pharmacology; D004926:Escherichia coli / Q000378:metabolism; D005786:Gene Expression Regulation; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007249:Inflammation / Q000378:metabolism*; D015850:Interleukin-6 / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology; D008738:Methyl Ethers / Q000494:pharmacology*; D010975:Platelet Aggregation Inhibitors / Q000494:pharmacology; D018805:Sepsis / Q000378:metabolism; D000077149:Sevoflurane; D051195:Toll-Like Receptor 2 / Q000378:metabolism*; D051197:Toll-Like Receptor 4 / Q000378:metabolism*; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
chemical_list: D003470:Culture Media; D015850:Interleukin-6; D008070:Lipopolysaccharides; D008738:Methyl Ethers; D010975:Platelet Aggregation Inhibitors; C495342:TLR2 protein, human; C495345:TLR4 protein, human; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D014409:Tumor Necrosis Factor-alpha; D000077149:Sevoflurane
references.citation: Eur J Anaesthesiol. 2009 Jul;26(7):597-602
references.pmid: 19522051
references.citation: J Immunol. 2011 Jan 15;186(2):1119-30
references.pmid: 21169547
references.citation: Clin Exp Immunol. 2009 Feb;155(2):224-30
references.pmid: 19032551
references.citation: Molecules. 2011 Dec 30;17(1):341-54
references.pmid: 22210172
references.citation: Anesthesiology. 2009 Jun;110(6):1271-8
references.pmid: 19417596
references.citation: Anesth Analg. 2005 Oct;101(4):1152-1156
references.pmid: 16192536
references.citation: J Cardiothorac Vasc Anesth. 2007 Aug;21(4):502-11
references.pmid: 17678775
references.citation: J Immunol. 1999 Jul 15;163(2):639-43
references.pmid: 10395652
references.citation: Eur J Pharmacol. 2010 Oct 25;645(1-3):39-46
references.pmid: 20655305
references.citation: Minerva Anestesiol. 2009 May;75(5):269-73
references.pmid: 18987572
references.citation: Eur Respir J. 2008 Jan;31(1):118-25
references.pmid: 17898018
references.citation: J Immunol. 2001 Feb 1;166(3):2018-24
references.pmid: 11160251
references.citation: Diabetes Metab Res Rev. 2010 Oct;26(7):550-8
references.pmid: 20818804
references.citation: Neurol Res. 2009 Mar;31(2):128-34
references.pmid: 19298752
references.citation: Br J Anaesth. 2004 Sep;93(3):422-7
references.pmid: 15277303
references.citation: J Biol Chem. 2000 Apr 14;275(15):11058-63
references.pmid: 10753909
references.citation: Acta Anaesthesiol Scand. 2007 Aug;51(7):893-9
references.pmid: 17635397
references.citation: CNS Neurosci Ther. 2011 Dec;17(6):605-11
references.pmid: 20875045
references.citation: Innate Immun. 2012 Feb;18(1):112-23
references.pmid: 21768203
references.citation: Mol Immunol. 2008 Nov;46(1):145-57
references.pmid: 18722665
references.citation: J Pathol. 2008 Jan;214(2):126-35
references.pmid: 18161748
references.citation: Immunobiology. 2007;212(9-10):715-22
references.pmid: 18086373
references.citation: Clin Exp Pharmacol Physiol. 2008 Sep;35(9):1043-51
references.pmid: 18505453
references.citation: Nature. 1998 Sep 17;395(6699):284-8
references.pmid: 9751057
references.citation: Intensive Care Med. 2010 Aug;36(8):1286-98
references.pmid: 20443110
references.citation: Biochem Pharmacol. 2006 Oct 30;72(9):1102-13
references.pmid: 16930560
references.citation: Crit Care. 2011;15(5):R241
references.pmid: 22011328
references.citation: Neurochem Int. 2011 Feb;58(3):337-43
references.pmid: 21163314
references.citation: Anesthesiology. 2007 Feb;106(2):262-8
references.pmid: 17264719
references.citation: Anesth Analg. 2007 Mar;104(3):638-45
references.pmid: 17312223
references.citation: Pharm Res. 2011 Nov;28(11):2910-9
references.pmid: 21786065
references.citation: Ann Card Anaesth. 2009 Jan-Jun;12(1):4-9
references.pmid: 19136748
references.citation: Curr Opin Anaesthesiol. 2007 Oct;20(5):395-9
references.pmid: 17873591
references.citation: Front Biosci (Elite Ed). 2011 Jan 01;3:604-15
references.pmid: 21196338
references.citation: Lab Anim. 2003 Jul;37(3):193-203
references.pmid: 12869281
references.citation: Int Immunopharmacol. 2002 Jan;2(1):95-104
references.pmid: 11789673
references.citation: Shock. 2008 Oct;30 Suppl 1:73-7
references.pmid: 18704004
references.citation: Anesth Analg. 2009 Nov;109(5):1591-7
references.pmid: 19843795
references.citation: Clin Diagn Lab Immunol. 2003 Jul;10(4):558-63
references.pmid: 12853386
references.citation: Am J Respir Cell Mol Biol. 2011 Sep;45(3):617-24
references.pmid: 21239603
references.citation: J Pathol. 2008 Jan;214(2):211-23
references.pmid: 18161754
references.citation: Blood. 2000 May 15;95(10):3044-51
references.pmid: 10807767
references.citation: Antioxid Redox Signal. 2004 Apr;6(2):439-48
references.pmid: 15025946
references.citation: J Anesth. 2005;19(2):150-6
references.pmid: 15875133