title: Hyperammonia induces specific liver injury through an intrinsic Ca2+-independent apoptosis pathway.
abstract: BACKGROUND
  Numerous pathological processes that affect liver function in patients with liver failure have been identified. Among them, hyperammonia is one of the most common phenomena.The purpose of this study was to determine whether hyperammonia could induced specific liver injury.
  
  
  METHODS
  Hyperammonemic cells were established using NH4Cl. The cells were assessed by MTT, ELISA, and flow cytometric analyses. The expression levels of selected genes and proteins were confirmed by quantitative RT-PCR and western blot analyses.
  
  
  RESULTS
  The effects of 20 mM NH4Cl pretreatment on the cell proliferation and apoptosis of primary hepatocytes and other cells were performed by MTT assays and flow cytometric analyses. Significant increasing in cytotoxicity and apoptosis were only observed in hepatocytes. The cell damage was reduced after adding BAPTA-AM but unchanged after adding EGTA. The expression levels of caspase-3, cytochrome C, calmodulin, and inducible nitric oxide synthase were increased and that of bcl-2 was reduced. The Na+-K+-ATPase activities in hyperammonia liver cells was no signiaficant difference compaired with the control group, but was decreased in astrocytes. NH4Cl pretreatment of primary hepatocytes promoted the activation of mitochondrial permeability transition pores and the mitochondria swelled irregularly.
  
  
  CONCLUSIONS
  Hyperammonia induces specific liver injury through an intrinsic Ca2+-independent apoptosis pathway.
authors.forename: Jingjing
authors.initials: J
authors.lastname: Li
authors.forename: Zujiang
authors.initials: Z
authors.lastname: Yu
authors.forename: Qiongye
authors.initials: Q
authors.lastname: Wang
authors.forename: Duolu
authors.initials: D
authors.lastname: Li
authors.forename: Bin
authors.initials: B
authors.lastname: Jia
authors.forename: Yubing
authors.initials: Y
authors.lastname: Zhou
authors.forename: Yanwei
authors.initials: Y
authors.lastname: Ye
authors.forename: Shen
authors.initials: S
authors.lastname: Shen
authors.forename: Yanfang
authors.initials: Y
authors.lastname: Wang
authors.forename: Shasha
authors.initials: S
authors.lastname: Li
authors.forename: Lu
authors.initials: L
authors.lastname: Bai
authors.affiliation: Department of Gastroenterology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China. Kanquancheng1963@163.com.
authors.forename: Quancheng
authors.initials: Q
authors.lastname: Kan
mesh_terms: D017209:Apoptosis; D002118:Calcium / Q000378:metabolism; D002147:Calmodulin / Q000235:genetics / Q000378:metabolism; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability; D049109:Cell Proliferation; D045304:Cytochromes c / Q000235:genetics / Q000378:metabolism; D020869:Gene Expression Profiling; D056945:Hep G2 Cells; D022781:Hepatocytes / Q000378:metabolism*; D006801:Humans; D022124:Hyperammonemia / Q000150:complications* / Q000378:metabolism; D008107:Liver Diseases / Q000209:etiology* / Q000378:metabolism; D061986:MCF-7 Cells; D008928:Mitochondria; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000032:analysis; D060888:Real-Time Polymerase Chain Reaction; D000254:Sodium-Potassium-Exchanging ATPase / Q000235:genetics / Q000378:metabolism
chemical_list: C000595707:BCL2 protein, human; D002147:Calmodulin; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D045304:Cytochromes c; C496319:NOS2 protein, human; D052247:Nitric Oxide Synthase Type II; C505407:CASP3 protein, human; D053148:Caspase 3; D000254:Sodium-Potassium-Exchanging ATPase; D002118:Calcium