title: P2X7 is involved in the anti-inflammation effects of levobupivacaine.
abstract: BACKGROUND
  We sough to elucidate whether purinergic P2X7 receptor is actively involved in the effects of levobupivacaine on inhibiting microglia activation.
  
  
  METHODS
  Microglia were treated with lipopolysaccharide (LPS, 50 ng/mL), LPS plus levobupivacaine (50 μM), or LPS plus levobupivacaine plus the P2X7 receptor agonist Bz-ATP (100 μM) and denoted as the LPS, LPS + Levo, and LPS + Levo + Bz-ATP group, respectively. Microglia activation was measured by assaying inflammatory molecules expression. Microglia activation was also measured by assaying neuronal cell viability using coculture of microglia and neurons, as activated microglia may cause neuron injury. We also measured the levels of P2X7 receptor activation in microglia using ethidium uptake assay.
  
  
  RESULTS
  Our data confirmed the effects of levobupivacaine on inhibiting inflammatory molecules upregulation in activated microglia, as the concentrations of interleukin (IL)-1β, tumor necrosis factor α, IL-6, and macrophage inflammatory protein 2, of the LPS + Levo group were significantly lower than those of the LPS group (all P < 0.05). Moreover, Bz-ATP significantly abrogated the inhibitory effects of levobupivacaine, as concentrations of IL-1β, tumor necrosis factor α, IL-6, and macrophage inflammatory protein 2 of the LPS + Levo + Bz-ATP group were significantly higher than those of the LPS + Levo group (all P < 0.05). In contrast, neuronal cell viability of the LPS + Levo group was significantly higher than those of the LPS and LPS + Levo + Bz-ATP groups (P = 0.012 and 0.002). Moreover, levels of P2X7 receptor activation of the LPS and LPS + Levo + Bz-ATP groups were significantly higher than that of the LPS + Levo group (P = 0.003 and 0.006).
  
  
  CONCLUSIONS
  P2X7 receptor is involved in the effects of levobupivacaine on inhibiting microglial activation.
authors.affiliation: Department of Anesthesiology, Mackay Memorial Hospital, Taipei, Taiwan; Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan.
authors.forename: Ya-Hsien
authors.initials: YH
authors.lastname: Huang
authors.affiliation: Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan.
authors.forename: Jiin-Cherng
authors.initials: JC
authors.lastname: Yen
authors.affiliation: Department of Surgery, Mackay Memorial Hospital, Taipei, Taiwan.
authors.forename: Jie-Jen
authors.initials: JJ
authors.lastname: Lee
authors.affiliation: Institute of Pharmacology, National Yang-Ming University, Taipei, Taiwan.
authors.forename: Jyh-Fei
authors.initials: JF
authors.lastname: Liao
authors.affiliation: Department of Anesthesiology, Tung's Taichung MetroHarbor Hospital, Taichung, Taiwan.
authors.forename: Wen-Jinn
authors.initials: WJ
authors.lastname: Liaw
authors.affiliation: Department of Anesthesiology, Taipei Tzu Chi Hospital, Taipei, Taiwan; School of Medicine, Tzu Chi University, Hualien, Taiwan. Electronic address: huangcj1112@gmail.com.
authors.forename: Chun-Jen
authors.initials: CJ
authors.lastname: Huang
mesh_terms: D000779:Anesthetics, Local / Q000494:pharmacology; D000818:Animals; D002045:Bupivacaine / Q000031:analogs & derivatives* / Q000494:pharmacology; D002450:Cell Communication / Q000187:drug effects / Q000276:immunology; D002461:Cell Line, Transformed; D002470:Cell Survival / Q000187:drug effects / Q000276:immunology; D018920:Coculture Techniques; D007249:Inflammation / Q000188:drug therapy* / Q000276:immunology*; D000077554:Levobupivacaine; D008070:Lipopolysaccharides / Q000494:pharmacology; D051379:Mice; D017628:Microglia / Q000166:cytology / Q000187:drug effects* / Q000276:immunology; D016328:NF-kappa B / Q000378:metabolism; D009474:Neurons / Q000166:cytology / Q000187:drug effects / Q000276:immunology; D058486:Receptors, Purinergic P2X7 / Q000276:immunology*
chemical_list: D000779:Anesthetics, Local; D008070:Lipopolysaccharides; D016328:NF-kappa B; D058486:Receptors, Purinergic P2X7; D000077554:Levobupivacaine; D002045:Bupivacaine
keywords: Cell viability; Levobupivacaine; Microglia; Purinergic P2X7 receptor