title: Urinary trypsin inhibitor attenuates LPS-induced endothelial barrier dysfunction by upregulation of vascular endothelial-cadherin expression.
abstract: BACKGROUND
  Urinary trypsin inhibitor (UTI) decreases inflammatory cytokine levels and mortality in experimental animal models of inflammation. Here, we observed the effect of UTI on lipopolysaccharide (LPS)-induced hyperpermeability in human umbilical vein endothelial cells (HUVECs) and explored the role of vascular endothelial-cadherin (VE-cadherin) in its effect.
  
  
  METHODS
  The effect of UTI on endothelial barrier hyperpermeability was detected by an electrical cell-substrate impedance sensing (ECIS) system and a transwell chamber system. The expression of VE-cadherin in HUVECs was examined by real-time PCR and western blot.
  
  
  RESULTS
  We demonstrated that the alleviation of LPS-induced barrier dysfunction could be achieved by pretreatment with 3000 U/mL of UTI. VE-cadherin monoclonal antibody (mAb) could inhibit the protective effects. UTI maintained VE-cadherin expression by increasing protein stability at both the transcriptional and post-transcriptional levels. Meanwhile, VE-cadherin expression on the cell surface increased when the cells were pretreated with UTI. Furthermore, pretreatment with UTI decreased the phosphorylation of VE-cadherin at Tyr658 but not Tyr731.
  
  
  CONCLUSIONS
  Our data show that prophylactic UTI maintains the endothelial barrier function, increases VE-cadherin expression, and inhibits the phosphorylation of VE-cadherin at Tyr658 under inflammatory conditions. It suggests a scientific and potential clinical therapeutic importance of UTI in treatment of inflammatory disorders.
authors.affiliation: Department of Emergency, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, 210029, People's Republic of China.
authors.forename: Jie
authors.initials: J
authors.lastname: Chen
authors.affiliation: The Laboratory of Neurotoxicology, School of Public Health, Nanjing Medical University, Nanjing, 210029, People's Republic of China.
authors.forename: Jun
authors.initials: J
authors.lastname: Wang
authors.affiliation: Department of Emergency, The Affiliated Hospital of Xuzhou Medical College, Xuzhou, 221000, People's Republic of China.
authors.forename: Chenglei
authors.initials: C
authors.lastname: Su
authors.affiliation: The Laboratory of Neurotoxicology, School of Public Health, Nanjing Medical University, Nanjing, 210029, People's Republic of China.
authors.forename: Wenyi
authors.initials: W
authors.lastname: Qian
authors.affiliation: Department of Basic Medical College, Nanjing Medical University, Nanjing, 210029, People's Republic of China.
authors.forename: Li
authors.initials: L
authors.lastname: Sun
authors.affiliation: Department of Emergency, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, 210029, People's Republic of China.
authors.forename: Hao
authors.initials: H
authors.lastname: Sun
authors.affiliation: Department of Emergency, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, 210029, People's Republic of China.
authors.forename: Junjie
authors.initials: J
authors.lastname: Chen
authors.affiliation: Department of Emergency, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, 210029, People's Republic of China.
authors.forename: Huazhong
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Department of Emergency, The First Affiliated Hospital of Nanjing Medical University, 300 Guangzhou Road, Nanjing, 210029, People's Republic of China. zhangjsojz@163.com.
authors.forename: Jinsong
authors.initials: J
authors.lastname: Zhang
mesh_terms: D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism*; D015820:Cadherins / Q000235:genetics / Q000378:metabolism*; D015815:Cell Adhesion Molecules / Q000235:genetics; D002463:Cell Membrane Permeability / Q000187:drug effects; D002465:Cell Movement / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D057167:Claudins / Q000235:genetics; D006023:Glycoproteins / Q000494:pharmacology*; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D008070:Lipopolysaccharides; D008264:Macrophages / Q000187:drug effects / Q000502:physiology; D012333:RNA, Messenger / Q000378:metabolism; D011956:Receptors, Cell Surface / Q000235:genetics; D015854:Up-Regulation; D000099152:Cadherin 5; D062765:Junctional Adhesion Molecule B
chemical_list: D015703:Antigens, CD; D015820:Cadherins; D015815:Cell Adhesion Molecules; D057167:Claudins; D006023:Glycoproteins; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D011956:Receptors, Cell Surface; D000099152:Cadherin 5; C407243:F11R protein, human; C411887:JAM2 protein, human; D062765:Junctional Adhesion Molecule B; C028665:urinastatin
keywords: Endothelial barrier dysfunction; Lipopolysaccharide; Urinary trypsin inhibitor; Vascular endothelial-cadherin