title: Local anesthetics inhibit substance P binding and evoked increases in intracellular Ca2+.
abstract: BACKGROUND
  During spinal and epidural anesthesia, local anesthetics reach concentrations in cerebrospinal fluid and spinal cord tissues at which their actions may extend beyond the classic blockade of sodium channels. This study examines the effects of several clinical and experimental local anesthetics on the binding and actions of a peptide neurotransmitter, substance P, known to be important in nociceptive transmission in the dorsal horn.
  
  
  METHODS
  The binding of radiolabeled (Bolton-Hunter modified) substance P was studied in chick brain membranes in the presence of local anesthetics. The increase in intracellular calcium [Ca2+]in evoked by substance P was measured by the fluorescent indicator fura-2 loaded in a murine cell line expressing substance P (NK1) receptors. Cells were preincubated with bupivacaine before and during the transient addition of substance P.
  
  
  RESULTS
  Both substance P binding and Ca2+ increase were inhibited half-maximally by approximately 1 mM bupivacaine at pH 7.5, whereas tetracaine, lidocaine, and benzocaine were slightly less potent at inhibiting binding. Concentration-dependent substance P-binding studies showed that bupivacaine's inhibition was not competitive. Inhibition of substance P binding by bupivacaine increased with increasing pH, but the protonated species appears to have some inhibitory activity, and quaternary lidocaine also inhibited binding. There was no stereoselectively to the binding inhibition.
  
  
  CONCLUSIONS
  Because millimolar concentrations of local anesthetics are within the range measured in spinal cord during intrathecal and epidural procedures, these results are consistent with a direct action of local anesthetics on tachykinin-mediated neurotransmission during regional anesthesia.
authors.affiliation: Department of Biological Chemistry, Harvard Medical School, Boston, Massachusetts 02115.
authors.forename: Y M
authors.initials: YM
authors.lastname: Li
authors.forename: D E
authors.initials: DE
authors.lastname: Wingrove
authors.forename: H P
authors.initials: HP
authors.lastname: Too
authors.forename: M
authors.initials: M
authors.lastname: Marnerakis
authors.forename: E R
authors.initials: ER
authors.lastname: Stimson
authors.forename: G R
authors.initials: GR
authors.lastname: Strichartz
authors.forename: J E
authors.initials: JE
authors.lastname: Maggio
mesh_terms: D000779:Anesthetics, Local / Q000494:pharmacology*; D000818:Animals; D001921:Brain / Q000187:drug effects* / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured / Q000187:drug effects / Q000378:metabolism; D002645:Chickens; D051379:Mice; D064729:Neurokinin-1 Receptor Antagonists; D018040:Receptors, Neurokinin-1 / Q000378:metabolism; D013237:Stereoisomerism; D013373:Substance P / Q000037:antagonists & inhibitors* / Q000378:metabolism
chemical_list: D000779:Anesthetics, Local; D064729:Neurokinin-1 Receptor Antagonists; D018040:Receptors, Neurokinin-1; D013373:Substance P; D002118:Calcium