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Role of Endotoxin in NF‐κB Activation by Ethanol in Rat Hepatocytes
Author(s) -
Matsumoto Hiroshi,
Sato Yasuhiro,
Azumi Junichi,
Kato Junji,
Niitsu Yoshiro,
Tamaki Keiji
Publication year - 2002
Publication title -
alcoholism: clinical and experimental research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.267
H-Index - 153
eISSN - 1530-0277
pISSN - 0145-6008
DOI - 10.1111/j.1530-0277.2002.tb02694.x
Subject(s) - ethanol , chemistry , nf κb , microbiology and biotechnology , biology , biochemistry , apoptosis
Background Endotoxin plays an important role in the progression of alcoholic liver injury. However, the role of endotoxin in acute activation of NF‐κB by ethanol remains unclear. Methods In primary rat hepatocyte cultures treated with ethanol and/or endotoxin, the DNA‐binding activity of NF‐κB in the nuclear extract was estimated by electrophoretic mobility shift assay. After pretreatment of a CYP2E1 inhibitor, 4‐methyl pyrazole or diallyl sulfide, NF‐κB activity was also measured in the same manner. Results Ethanol or endotoxin caused activation of NF‐κB in primary rat hepatocytes. Taking 50 mM of ethanol and endotoxin together raised the rapid increase in NF‐κB activation, but both 100 mM of ethanol and endotoxin treatment reduced the increase. Addition of diallyl sulfide decreased the activity at rapid phase but increased it after 60 min, whereas addition of 4‐methyl pyrazole caused no change of the activity at rapid phase but raised it after 60 min. Pretreatment with both endotoxin and a CYP2E1 inhibitor raised the activation of NF‐κB constantly after ethanol addition. These findings suggest that endotoxin plays a critical role in the metabolism‐independent activation of NF‐κB by ethanol. Conclusions Endotoxin raised metabolism‐independent activation of NF‐κB by high ethanol in rat hepatocytes.