title: Plasminogen activator inhibitor-1 regulates LPS induced inflammation in rat macrophages through autophagy activation.
abstract: BACKGROUND
  The mechanisms by which plasminogen activator inhibitor-1 (PAI-1) regulates inflammation, especially in acute respiratory distress syndrome (ARDS), are largely unknown.
  
  
  OBJECTIVE
  To assess the relationship between PAI-1 and autophagy in inflammatory reactions induced by LPS in rat NR8383 cells.
  
  
  METHODS
  ELISA was used to assess the amounts of TNF-α, IL-1β, and PAI-1 in cell culture supernatants; TLR4, MyD88, PAI-1, LC3, Beclin1, and mTOR protein and mRNA levels were determined by western blot and quantitative RT-PCR, respectively; western blot was used to determine NF-κB protein levels. To further evaluate the role of PAI-1, the PAI-1 gene was downregulated and overexpressed using the siRNA transfection technology and the pCDH-PAI-1, respectively. Finally, the GFP Positive Expression Rate Method was used to determine the rate of GFP-LC3 positive NR8383 cells.
  
  
  RESULTS
  In LPS-induced NR8383 cells, TNF-α, IL-1β, and PAI-1 expression levels increased remarkably. Upon PAI-1 knockdown, TNF-α, IL-1β, PAI-1, TLR4, MyD88, NF-κB, LC3, and Beclin1 levels were decreased, while mTOR increased. Conversely, overexpression of PAI-1 resulted in increased amounts of TNF-α, IL-1β, PAI-1, TLR4, MyD88, NF-κB, LC3, and Beclin1. However, no significant change was observed in mTOR expression.
  
  
  CONCLUSIONS
  In NR8383 cells, PAI-1 contributes in the regulation of LPS-induced inflammation, likely by promoting autophagy.
authors.affiliation: Department of Pulmonary Medicine, Zhongshan Hospital, Fudan University, No. 180, Feng Lin Road, Shanghai 200032, China.
authors.forename: Zhong-Hui
authors.initials: ZH
authors.lastname: Wang
authors.affiliation: Department of Geriatrics, Zhongshan Hospital, Fudan University, No. 180, Feng Lin Road, Shanghai 200032, China.
authors.forename: Wei-Ying
authors.initials: WY
authors.lastname: Ren
authors.affiliation: Department of Pulmonary Medicine, Zhongshan Hospital, Fudan University, No. 180, Feng Lin Road, Shanghai 200032, China.
authors.forename: Lei
authors.initials: L
authors.lastname: Zhu
authors.affiliation: Department of Pulmonary Medicine, Zhongshan Hospital, Fudan University, No. 180, Feng Lin Road, Shanghai 200032, China.
authors.forename: Li-Juan
authors.initials: LJ
authors.lastname: Hu
mesh_terms: D000818:Animals; D051017:Apoptosis Regulatory Proteins / Q000235:genetics / Q000378:metabolism; D001343:Autophagy; D000071186:Beclin-1; D002460:Cell Line; D007249:Inflammation / Q000378:metabolism; D053583:Interleukin-1beta / Q000235:genetics / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D008869:Microtubule-Associated Proteins / Q000235:genetics / Q000378:metabolism; D053594:Myeloid Differentiation Factor 88 / Q000235:genetics / Q000378:metabolism; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D017395:Plasminogen Activator Inhibitor 1 / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D058570:TOR Serine-Threonine Kinases / Q000235:genetics / Q000378:metabolism; D051197:Toll-Like Receptor 4 / Q000819:agonists / Q000235:genetics / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism
chemical_list: D051017:Apoptosis Regulatory Proteins; D000071186:Beclin-1; D053583:Interleukin-1beta; D008070:Lipopolysaccharides; D008869:Microtubule-Associated Proteins; D053594:Myeloid Differentiation Factor 88; D016328:NF-kappa B; D017395:Plasminogen Activator Inhibitor 1; D012333:RNA, Messenger; D058570:TOR Serine-Threonine Kinases; D051197:Toll-Like Receptor 4; D014409:Tumor Necrosis Factor-alpha; C512829:Becn1 protein, rat; C526109:Anxa3 protein, rat