title: Penehyclidine hydrochloride inhibits the release of high-mobility group box 1 in lipopolysaccharide-activated RAW264.7 cells and cecal ligation and puncture-induced septic mice.
abstract: BACKGROUND
  High-mobility group box 1 (HMGB1) is a critical mediator in the pathogenesis of many inflammatory diseases. Penehyclidine hydrochloride (PHC) has been proven to reduce sepsis-related mortality and sepsis-induced pathological complications. These effects are because of the reduced expression and release of many inflammatory mediators, although it is not clear whether PHC affects the expression and release of HMGB1. In this study, we explored the effect of PHC on the release of HMGB1 in lipopolysaccharide (LPS)-activated RAW264.7 cells and cecal ligation and puncture (CLP)-induced septic mice.
  
  
  METHODS
  RAW264.7 cells were incubated with LPS in the presence or absence of various concentrations of PHC. The expression levels of HMGB1 in the culture supernatant were detected by enzyme-linked immunosorbent assay and real-time polymerase chain reaction. Western blotting was used to observe changes in the translocation of HMGB1 from the nucleus to the cytoplasm, and the nuclear factor (NF)-κB activity in the nuclear extract was detected by the NF-κB p50/p65 Transcription Factor Assay Kit. In addition, 48 CLP-induced septic BALB/c were treated with different concentrations of PHC 1 h before performing the CLP, and the level of serum HMGB1 and the functional parameters of multiple organs were determined using several detection kits.
  
  
  RESULTS
  We found that PHC inhibited the release of HMGB1 in LPS-activated RAW264.7 cells and CLP-induced septic mice. PHC inhibited the translocation of HMGB1 from the nucleus to the cytoplasm and also suppressed the expression of HMGB1 messenger RNA. Furthermore, PHC inhibited the translocation of NF-κB from the cytoplasm to the nucleus in LPS-activated RAW264.7 cells in a dose-dependent manner. Compared with the CLP alone group, the levels of alanine aminotransferase, aspartate aminotransferase, blood urea nitrogen, creatinine, and creatine kinase were significantly decreased in mice treated with 0.45 mg/kg of PHC (P < 0.01).
  
  
  CONCLUSIONS
  Our study demonstrates that PHC inhibits the translocation of HMGB1 from the nucleus to the cytoplasm and the expression of HMGB1 messenger RNA in a dose-dependent manner. The mechanism responsible for these effects involves the NF-κB signaling pathway. Moreover, PHC can significantly protect important organs, such as the liver, kidney, and heart in mice with sepsis.
authors.affiliation: Department of Anesthesiology, Cangzhou Central Hospital, Hebei Medical University, Hebei, China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Yang
authors.forename: Xiangge
authors.initials: X
authors.lastname: Liu
authors.forename: Zhongyan
authors.initials: Z
authors.lastname: Yao
authors.forename: Shunhong
authors.initials: S
authors.lastname: Mao
authors.forename: Qianjie
authors.initials: Q
authors.lastname: Wei
authors.forename: Yulin
authors.initials: Y
authors.lastname: Chang
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002432:Cecum; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D018680:Cholinergic Antagonists / Q000494:pharmacology*; D024243:HMGB1 Protein / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008026:Ligation; D008070:Lipopolysaccharides / Q000494:pharmacology; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D016328:NF-kappa B / Q000378:metabolism; D021381:Protein Transport / Q000187:drug effects; D011812:Quinuclidines / Q000494:pharmacology*; D018805:Sepsis / Q000188:drug therapy* / Q000378:metabolism
chemical_list: D018680:Cholinergic Antagonists; D024243:HMGB1 Protein; D008070:Lipopolysaccharides; D016328:NF-kappa B; D011812:Quinuclidines; C486140:penehyclidine
keywords: HMGB1 protein; Lipopolysaccharides; NF-κB; Penehyclidine hydrochloride; Sepsis