title: Vaspin inhibited proinflammatory cytokine induced activation of nuclear factor-kappa B and its downstream molecules in human endothelial EA.hy926 cells.
abstract: OBJECTIVE
  In this study, we investigated the effects of visceral adipose tissue-derived serpin (vaspin), a newly discovered adipocytokine, on nuclear factor-kappa B (NF-κB) and its downstream molecules in proinflammatory cytokines, tumor necrosis factor-α (TNF-α) and interleukine-1 (IL-1), stimulated human endothelial EA.hy926 cells to elucidate the role of vaspin in the inflammatory states of endothelium.
  
  
  METHODS
  A NF-κB luciferase reporter system was constructed and stably transfected into human endothelial cell line EA.hy926. Following transfection, EA.hy926 cells were pretreated with various concentrations of vaspin (0-320 ng/ml) before TNF-α and IL-1 stimulation. The transcription activity of NF-κB was determined using luciferase reporter assay. Expression levels of NF-κB downstream inflammatory cytokines, TNF-α, IL-1 and IL-6 were measured by enzyme-linked immunosorbent assay (ELISA). Expressions of adhesion molecules and chemokines, intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1) and monocyte chemotactic protein-1 (MCP-1) were determined by quantitative real-time PCR (RT-PCR) and western blot in mRNA and protein levels, respectively.
  
  
  RESULTS
  Results showed that vaspin inhibited TNF-α and IL-1 mediated activation of NF-κB and its downstream molecules in a concentration-dependent manner (P<0.05).
  
  
  CONCLUSIONS
  We conclude that vaspin protected endothelial cells from proinflammatory cytokines induced inflammation by inhibition of NF-κB and its downstream molecules.
authors.affiliation: Department of Endocrinology, Shanxi DAYI Hospital, Shanxi Medical University, Taiyuan 030032, China. Electronic address: lswspring6@gmail.com.
authors.forename: Shiwei
authors.initials: S
authors.lastname: Liu
authors.affiliation: Department of Biochemistry, Shanxi Medical University, Taiyuan 030001, China.
authors.forename: Yanting
authors.initials: Y
authors.lastname: Dong
authors.affiliation: Department of Medical Statistics, School of Public Health of Shanxi Medical University, Taiyuan 030001, China.
authors.forename: Tong
authors.initials: T
authors.lastname: Wang
authors.affiliation: Shanxi DAYI Hospital, Shanxi Medical University, Taiyuan 030032, China.
authors.forename: Shujun
authors.initials: S
authors.lastname: Zhao
authors.affiliation: Department of Endocrinology, Shanxi DAYI Hospital, Shanxi Medical University, Taiyuan 030032, China.
authors.forename: Kun
authors.initials: K
authors.lastname: Yang
authors.affiliation: Department of Endocrinology, Shanxi DAYI Hospital, Shanxi Medical University, Taiyuan 030032, China.
authors.forename: Xiaoqin
authors.initials: X
authors.lastname: Chen
authors.affiliation: Department of Endocrinology, Shanxi DAYI Hospital, Shanxi Medical University, Taiyuan 030032, China.
authors.forename: Caihong
authors.initials: C
authors.lastname: Zheng
mesh_terms: D015153:Blotting, Western; D002478:Cells, Cultured; D018932:Chemokine CCL2 / Q000235:genetics / Q000378:metabolism*; D016207:Cytokines / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000187:drug effects* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D007249:Inflammation / Q000188:drug therapy* / Q000235:genetics / Q000378:metabolism; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism*; D007375:Interleukin-1 / Q000494:pharmacology; D015850:Interleukin-6 / Q000494:pharmacology; D008156:Luciferases / Q000378:metabolism; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D015843:Serpins / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology; D019010:Vascular Cell Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism*
chemical_list: C111835:CCL2 protein, human; D018932:Chemokine CCL2; D016207:Cytokines; C573420:ICAM1 protein, human; D007375:Interleukin-1; D015850:Interleukin-6; D016328:NF-kappa B; D012333:RNA, Messenger; C507339:SERPINA12 protein, human; D015843:Serpins; D014409:Tumor Necrosis Factor-alpha; D019010:Vascular Cell Adhesion Molecule-1; D018799:Intercellular Adhesion Molecule-1; D008156:Luciferases
keywords: Endothelium; Inflammation; Insulin resistance; NF-κB; Vaspin