title: Hypothermia-induced loss of endothelial barrier function is restored after dopamine pretreatment: role of p42/p44 activation.
abstract: BACKGROUND
  Donor dopamine usage is associated with improved immediate graft function after renal transplantation. Although prolonged cold preservation results in an increased vascular permeability, the present study was conducted to examine in vitro and in vivo if dopamine pretreatment influences endothelial barrier function under such conditions.
  
  
  METHODS
  To assess cold preservation injury in vitro and in vivo, cultured human umbilical vein endothelial cells (HUVEC) and Lewis donor rats were pretreated with dopamine or isotonic saline prior to cold storage. Injury was determined by lactate dehydrogenase (LDH) release, histology, and functional analysis.
  
  
  RESULTS
  In vitro cold storage resulted in intercellular gap formation in both untreated and dopamine pretreated HUVEC. In the latter monolayer integrity was completely restored upon rewarming and paracellular transport of fluorescein isothiocyanate-dextran was significantly reduced. In dopamine treated HUVEC, intercellular gap formation was independent of cell death and was associated with redistribution of junctional proteins and condensation of cytoskeleton proteins. In untreated HUVEC proteolysis and cell death were clearly evident after hypothermia. Closing of intercellular gaps was dependent on p42/p44 activation. Regeneration of adenosine triphosphate was only observed in dopamine pretreated cells. Only in dopamine treated Lewis renal allografts subjected to cold storage, activation of p42/p44 occurred upon rewarming. These grafts had a better renal function and displayed less inflammatory cells five days after transplantation.
  
  
  CONCLUSIONS
  Our study demonstrates beneficial effects of dopamine treatment on cold storage induced endothelial barrier disturbances. This may contribute to the positive effects of catecholamines on immediate graft function of renal allografts in men.
authors.affiliation: Department of Medicine (Nephrology), University Hospital Mannheim, University of Heidelberg, Germany.
authors.forename: Paul-Thomas
authors.initials: PT
authors.lastname: Brinkkoetter
authors.forename: Grietje C
authors.initials: GC
authors.lastname: Beck
authors.forename: Uwe
authors.initials: U
authors.lastname: Gottmann
authors.forename: Ralf
authors.initials: R
authors.lastname: Loesel
authors.forename: Ulf
authors.initials: U
authors.lastname: Schnetzke
authors.forename: Boris
authors.initials: B
authors.lastname: Rudic
authors.forename: Christine
authors.initials: C
authors.lastname: Hanusch
authors.forename: Neysan
authors.initials: N
authors.lastname: Rafat
authors.forename: Zhenzi
authors.initials: Z
authors.lastname: Liu
authors.forename: Christel
authors.initials: C
authors.lastname: Weiss
authors.forename: Henri G D
authors.initials: HG
authors.lastname: Leuvinik
authors.forename: Rutger
authors.initials: R
authors.lastname: Ploeg
authors.forename: Claude
authors.initials: C
authors.lastname: Braun
authors.forename: Peter
authors.initials: P
authors.lastname: Schnuelle
authors.forename: Fokko J
authors.initials: FJ
authors.lastname: van der Woude
authors.forename: Benito A
authors.initials: BA
authors.lastname: Yard
mesh_terms: D002478:Cells, Cultured; D003080:Cold Temperature; D004298:Dopamine / Q000494:pharmacology*; D042783:Endothelial Cells / Q000502:physiology*; D004789:Enzyme Activation; D006801:Humans; D019950:Mitogen-Activated Protein Kinase 1 / Q000502:physiology*; D048052:Mitogen-Activated Protein Kinase 3 / Q000502:physiology*; D009926:Organ Preservation; D014404:Tubulin / Q000032:analysis; D014746:Vimentin / Q000032:analysis
chemical_list: D014404:Tubulin; D014746:Vimentin; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D004298:Dopamine