title: Endothelial barrier protection by local anesthetics: ropivacaine and lidocaine block tumor necrosis factor-α-induced endothelial cell Src activation.
abstract: BACKGROUND
  Pulmonary endothelial barrier dysfunction mediated in part by Src-kinase activation plays a crucial role in acute inflammatory disease. Proinflammatory cytokines, such as tumor necrosis factor-α (TNFα), activate Src via phosphatidylinositide 3-kinase/Akt-dependent nitric oxide generation, a process initiated by recruitment of phosphatidylinositide 3-kinase regulatory subunit p85 to TNF-receptor-1. Because amide-linked local anesthetics have well-established anti-inflammatory effects, the authors hypothesized that ropivacaine and lidocaine attenuate inflammatory Src signaling by disrupting the phosphatidylinositide 3-kinase-Akt-nitric oxide pathway, thus blocking Src-dependent neutrophil adhesion and endothelial hyperpermeability.
  
  
  METHODS
  Human lung microvascular endothelial cells, incubated with TNFα in the absence or presence of clinically relevant concentrations of ropivacaine and lidocaine, were analyzed by Western blot, probing for phosphorylated/activated Src, endothelial nitric oxide synthase, Akt, intercellular adhesion molecule-1, and caveolin-1. The effect of ropivacaine on TNFα-induced nitric oxide generation, co-immunoprecipitation of TNF-receptor-1 with p85, neutrophil adhesion, and endothelial barrier disruption were assessed.
  
  
  RESULTS
  Ropivacaine and lidocaine attenuated TNFα-induced Src activation (half-maximal inhibitory concentration [IC50] = 8.611 × 10 M for ropivacaine; IC50 = 5.864 × 10 M for lidocaine) and endothelial nitric oxide synthase phosphorylation (IC50 = 7.572 × 10 M for ropivacaine; IC50 = 6.377 × 10 M for lidocaine). Akt activation (n = 7; P = 0.006) and stimulus-dependent binding of TNF-receptor-1 and p85 (n = 6; P = 0.043) were blocked by 1 nM of ropivacaine. TNFα-induced neutrophil adhesion and disruption of endothelial monolayers via Src-dependent intercellular adhesion molecule-1- and caveolin-1-phosphorylation, respectively, were also attenuated.
  
  
  CONCLUSIONS
  Ropivacaine and lidocaine effectively blocked inflammatory TNFα signaling in endothelial cells by attenuating p85 recruitment to TNF-receptor-1. The resultant decrease in Akt, endothelial nitric oxide synthase, and Src phosphorylation reduced neutrophil adhesion and endothelial hyperpermeability. This novel anti-inflammatory "side-effect" of ropivacaine and lidocaine may provide therapeutic benefit in acute inflammatory disease.
authors: From the Department of Anesthesiology (T.P., E.G.V.-V., D.E.S., R.D.M.), Department of Pharmacology (F.R.B., M.M., G.C., M.G.B., R.D.M.), Department of Medicine (M.G.B.), and Center for Lung and Vascular Biology (R.D.M.), University of Illinois Hospital and Health Sciences System, Chicago, Illinois; Institute of Anesthesiology (T.P.), University Hospital Zurich, Zurich, Switzerland; Institute of Physiology, Zurich Center for Integrative Human Physiology (B.B.-S.), University of Zurich; Department of Anesthesiology (A.B.), Balgrist University Hospital Zurich, Zurich, Switzerland; and Department of Anesthesiology (E.G.V.-V.), Jesse Brown VA Medical Center, Chicago, Illinois.
authors: Tobias
authors: T
authors: Piegeler
authors: E Gina
authors: EG
authors: Votta-Velis
authors: Farnaz R
authors: FR
authors: Bakhshi
authors: Mao
authors: M
authors: Mao
authors: Graeme
authors: G
authors: Carnegie
authors: Marcelo G
authors: MG
authors: Bonini
authors: David E
authors: DE
authors: Schwartz
authors: Alain
authors: A
authors: Borgeat
authors: Beatrice
authors: B
authors: Beck-Schimmer
authors: Richard D
authors: RD
authors: Minshall
mesh_terms: D000577:Amides / Q000494:pharmacology*; D000779:Anesthetics, Local / Q000494:pharmacology*; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D004789:Enzyme Activation / Q000187:drug effects / Q000502:physiology; D006801:Humans; D008012:Lidocaine / Q000494:pharmacology*; D008168:Lung / Q000187:drug effects / Q000201:enzymology; D008833:Microcirculation / Q000187:drug effects / Q000502:physiology; D000077212:Ropivacaine; D014409:Tumor Necrosis Factor-alpha / Q000008:administration & dosage / Q000037:antagonists & inhibitors*; D019061:src-Family Kinases / Q000378:metabolism* / Q000502:physiology
chemical_list: D000577:Amides; D000779:Anesthetics, Local; D014409:Tumor Necrosis Factor-alpha; D000077212:Ropivacaine; D008012:Lidocaine; D019061:src-Family Kinases