title: Effect of Lysosomotropic Polyamineoxidase Inhibitor MDL-72527 on Platelet Activation.
abstract: OBJECTIVE
  The polyamine oxidase inhibitor MDL-72527 (N1,N4-bis(2,3-butadienyl)-1,4-butanediamine) were expected to increase the abundance of spermine, a powerful inhibitor of platelet activation. Nothing is known, however, on the sensitivity of platelet function and survival to MDL-72527 exposure. The present study thus explored whether MDL-72527 modifies function and survival of platelets without and with platelet activation by collagen related peptide (CRP).
  
  
  METHODS
  Platelets isolated from wild-type mice were exposed for 30 minutes to MDL-72527 (100 µM) with or without subsequent activation with CRP (2-5 µg/ml). Flow cytometry was employed to estimate cytosolic Ca2+-activity ([Ca2+]i) from Fluo-3 fluorescence, platelet degranulation from P-selectin abundance, integrin activation from αIIbβ3 integrin abundance, generation of reactive oxygen species (ROS) from DCFDA fluorescence, phospholipid scrambling of the cell membrane from annexin-V-binding, platelet volume from forward scatter and aggregation utilizing staining with CD9-APC and CD9-PE.
  
  
  RESULTS
  In the absence of CRP, exposure of platelets to MDL-72527 did not significantly modify [Ca2+]i, P-selectin abundance, αIIbβ3 integrin abundance, ROS, annexin-V-binding, and forward scatter. The addition of 2-5 µg/ml CRP was followed by significant increase of [Ca2+]i, P-selectin abundance, αIIbβ3 integrin activation, ROS abundance, annexin-V-binding, and aggregation as well as a significant decrease of forward scatter, all effects significantly blunted or virtually abolished in the presence of MDL-72527.
  
  
  CONCLUSIONS
  MDL-72527 is a powerful inhibitor of platelet activation, apoptosis and aggregation.
authors.forename: Guoxing
authors.initials: G
authors.lastname: Liu
authors.forename: Hang
authors.initials: H
authors.lastname: Cao
authors.forename: Guilai
authors.initials: G
authors.lastname: Liu
authors.forename: David
authors.initials: D
authors.lastname: Heinzmann
authors.forename: Hong
authors.initials: H
authors.lastname: Chen
authors.forename: Anja T
authors.initials: AT
authors.lastname: Umbach
authors.forename: Meinrad
authors.initials: M
authors.lastname: Gawaz
authors.forename: Florian
authors.initials: F
authors.lastname: Lang
mesh_terms: D000814:Aniline Compounds / Q000737:chemistry; D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D001792:Blood Platelets / Q000166:cytology / Q000378:metabolism; D002118:Calcium / Q000737:chemistry / Q000378:metabolism; D002352:Carrier Proteins / Q000494:pharmacology; D002462:Cell Membrane / Q000378:metabolism; D005260:Female; D005434:Flow Cytometry; D008297:Male; D051379:Mice; D000587:Oxidoreductases Acting on CH-NH Group Donors / Q000037:antagonists & inhibitors / Q000378:metabolism; D019007:P-Selectin / Q000378:metabolism; D010455:Peptides / Q000494:pharmacology; D010718:Phosphatidylserines; D015539:Platelet Activation / Q000187:drug effects*; D019039:Platelet Glycoprotein GPIIb-IIIa Complex / Q000378:metabolism; D011700:Putrescine / Q000031:analogs & derivatives* / Q000494:pharmacology; D017382:Reactive Oxygen Species / Q000378:metabolism; D014966:Xanthenes / Q000737:chemistry; D000097667:Polyamine Oxidase
chemical_list: D000814:Aniline Compounds; D002118:Calcium; D002352:Carrier Proteins; D000587:Oxidoreductases Acting on CH-NH Group Donors; D019007:P-Selectin; D010455:Peptides; D010718:Phosphatidylserines; D019039:Platelet Glycoprotein GPIIb-IIIa Complex; D011700:Putrescine; D017382:Reactive Oxygen Species; D014966:Xanthenes; D000097667:Polyamine Oxidase; C435908:collagen-related peptide; C044445:MDL 72527; C059715:Fluo-3