title: Effect of acidosis on IL-8 and MCP-1 during hypoxia and reoxygenation in human NT2-N neurons.
abstract: Inflammation probably plays a significant role in perinatal brain injury. To study the contribution of locally produced cytokines, the effect on cell death of addition of IL-8 and MCP-1 or antibodies to these, and the impact of acidosis, human postmitotic NT2-N neurons were exposed to 3 h of hypoxia and glucose deprivation and reoxygenated for 21 h. After 3 h of hypoxia with neutral medium, IL-8 was significantly increased compared to controls (150 (100-250)% vs. 100 (85-115)%, p=0.023). After 21 h of neutral reoxygenation, both IL-8 (380 (110-710)% vs. 150 (85-260)%, p=0.041) and monocyte chemoattractant protein-1 (MCP-1) (650 (440-2000)% vs. 310 (230-340)%, p=0.007) were significantly increased compared to controls. After 3 h of hypoxia, both IL-8 (p=0.002) and MCP-1 (p=0.008) were significantly lower in cells with acidotic compared with cells with neutral medium. Acidosis during reoxygenation, however, significantly increased IL-8 release, whereas MCP-1 release was diminished. Similar effects of acidosis were seen in normoxic controls. The cells also secreted RANTES and IP-10, but not 8 other cytokines tested. We found no effect on cell death, measured by MTT assay, of addition of IL-8, MCP-1 or antibodies to these. We conclude that human NT2-N neurons release IL-8 and MCP-1 during 21 h of reoxygenation after 3 h of hypoxia. Acidosis led to a differential effect on IL-8 and MCP-1, with increased IL-8 and decreased MCP-1, both during reoxygenation and in normoxic controls. IL-8 and MCP-1 had no effect on cell death.
authors.affiliation: Department of Pediatric Research, Rikshospitalet-Radiumhospitalet Medical Center and University of Oslo, N-0027 Oslo, Norway. elisabeth.froyland@medisin.uio.no
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Frøyland
authors.forename: Elena Didenko
authors.initials: ED
authors.lastname: Pedersen
authors.forename: Anne-Katrine
authors.initials: AK
authors.lastname: Kvissel
authors.forename: Runar
authors.initials: R
authors.lastname: Almaas
authors.forename: Anne
authors.initials: A
authors.lastname: Pharo
authors.forename: Bjørn Steen
authors.initials: BS
authors.lastname: Skålhegg
authors.forename: Tom Eirik
authors.initials: TE
authors.lastname: Mollnes
authors.forename: Terje
authors.initials: T
authors.lastname: Rootwelt
mesh_terms: D000138:Acidosis / Q000378:metabolism*; D000906:Antibodies / Q000494:pharmacology; D015687:Cell Hypoxia / Q000502:physiology; D002460:Cell Line; D018932:Chemokine CCL2 / Q000276:immunology / Q000378:metabolism*; D018946:Chemokine CCL5 / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005455:Fluorescent Antibody Technique / Q000379:methods; D005786:Gene Expression Regulation / Q000187:drug effects*; D005947:Glucose / Q000172:deficiency; D006801:Humans; D000860:Hypoxia; D016209:Interleukin-8 / Q000276:immunology / Q000378:metabolism*; D016900:Neurofilament Proteins / Q000378:metabolism; D009474:Neurons / Q000502:physiology*; D010100:Oxygen / Q000008:administration & dosage*; D018709:Statistics, Nonparametric; D013778:Tetrazolium Salts; D013844:Thiazoles; D013997:Time Factors
chemical_list: D000906:Antibodies; D018932:Chemokine CCL2; D018946:Chemokine CCL5; D016209:Interleukin-8; D016900:Neurofilament Proteins; D013778:Tetrazolium Salts; D013844:Thiazoles; C051774:neurofilament protein H; C022616:thiazolyl blue; D005947:Glucose; D010100:Oxygen