title: Inhibition of peptidylarginine deiminase attenuates inflammation and improves survival in a rat model of hemorrhagic shock.
abstract: BACKGROUND
  We have recently shown that inhibition of peptidylarginine deiminase (PAD) improves survival in a rodent model of lethal cecal ligation and puncture. The roles of PAD inhibitors in hemorrhagic shock (HS), however, are largely unknown. The goal of this study was to investigate the effects of YW3-56, a novel PAD inhibitor, on survival after severe HS.
  
  
  METHODS
  Mouse macrophages were exposed to hypoxic conditions followed by reoxygenation in the presence or absence of YW3-56. Enzyme-linked immunosorbent assay (ELISA) was performed to measure levels of secreted tumor necrosis factor α and interleukin-6 in the culture medium. Cell viability was determined by methyl thiazolyl tetrazolium assay. In the survival experiment, anesthetized male Wistar-Kyoto rats (n = 10/group) were subjected to 55% blood loss, and treated with or without YW3-56 (10 mg/kg, intraperitoneally). Survival was monitored for 12 h. In the nonsurvival experiment, morphologic changes of the lungs were examined. Levels of circulating cytokine-induced neutrophil chemoattractant 1 (CINC-1) and myeloperoxidase (MPO) in the lungs were measured by ELISA. Expression of lung intercellular adhesion molecules-1 (ICAM-1) was also determined by Western blotting.
  
  
  RESULTS
  Hypoxia/reoxygenation (H/R) insult induced tumor necrosis factor α and interleukin-6 secretion from macrophages, which was significantly attenuated by YW3-56 treatment. YW3-56 treatment also increased cell viability when macrophages were exposed to H/R up to 6/15 h and improved survival rate from 20% to 60% in lethal HS rat model. Compared to the sham groups, pulmonary MPO activity and ICAM-1 expression in the HS group were significantly increased, and acute lung injury was associated with a higher degree of CINC-1 levels in serum. Intraperitoneal delivery of YW3-56 significantly reduced pulmonary MPO and ICAM-1 expression and attenuated acute lung injury.
  
  
  CONCLUSIONS
  Our results demonstrate for the first time that administration of YW3-56, a novel PAD inhibitor, can improve survival in a rat model of HS and in a cell culture model of H/R. The survival advantage is associated with an attenuation of local and systemic pro-inflammatory cytokines and the protection against acute lung injury after hemorrhage. Thus, PAD inhibition may represent a novel and promising therapeutic strategy for severe HS.
authors.affiliation: Department of Surgery, Zhongda Hospital, School of Medicine, Southeast University, Nanjing, China; Department of Surgery, University of Michigan, Ann Arbor, Michigan.
authors.forename: Wei
authors.initials: W
authors.lastname: He
authors.affiliation: Department of Surgery, University of Michigan, Ann Arbor, Michigan; Harvard University, Cambridge, Massachusetts.
authors.forename: Peter
authors.initials: P
authors.lastname: Zhou
authors.affiliation: Department of Surgery, University of Michigan, Ann Arbor, Michigan; Department of Surgical ICU, Beijing Hospital Ministry of Health, Beijing, China.
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Chang
authors.affiliation: Department of Surgery, University of Michigan, Ann Arbor, Michigan.
authors.forename: Baoling
authors.initials: B
authors.lastname: Liu
authors.affiliation: Department of Systems Leadership and Effectiveness Science, University of Michigan School of Nursing, Ann Arbor, Michigan.
authors.forename: Xuefeng
authors.initials: X
authors.lastname: Liu
authors.affiliation: Department of Biochemistry and Molecular Biology, Pennsylvania State University, Pennsylvania.
authors.forename: Yanming
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Surgery, University of Michigan, Ann Arbor, Michigan.
authors.forename: Yongqing
authors.initials: Y
authors.lastname: Li
authors.affiliation: Department of Surgery, University of Michigan, Ann Arbor, Michigan. Electronic address: alamh@med.umich.edu.
authors.forename: Hasan B
authors.initials: HB
authors.lastname: Alam
mesh_terms: D015081:2-Naphthylamine / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000627:therapeutic use; D000818:Animals; D001120:Arginine / Q000031:analogs & derivatives* / Q000494:pharmacology / Q000627:therapeutic use; D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology / Q000627:therapeutic use*; D004797:Enzyme-Linked Immunosorbent Assay; D006867:Hydrolases / Q000037:antagonists & inhibitors*; D007274:Injections, Intraperitoneal; D008168:Lung / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D000076342:Protein-Arginine Deiminases; D011897:Random Allocation; D051381:Rats; D011921:Rats, Inbred WKY; D017207:Rats, Sprague-Dawley; D015427:Reperfusion Injury / Q000188:drug therapy / Q000378:metabolism; D012771:Shock, Hemorrhagic / Q000188:drug therapy* / Q000378:metabolism / Q000401:mortality
chemical_list: D015415:Biomarkers; D016207:Cytokines; D004791:Enzyme Inhibitors; C000607342:YW3-56; D001120:Arginine; D015081:2-Naphthylamine; D006867:Hydrolases; D000076342:Protein-Arginine Deiminases
keywords: Hemorrhagic shock; Inflammation; Peptidylarginine deiminase inhibitor; Rat; Survival