J Korean Med Sci.  2002 Apr;17(2):193-200. 10.3346/jkms.2002.17.2.193.

NF-kappaB Binding Activity and Cyclooxygenase-2 Expression in Persistent betaCCI(4)-Treated Rat Liver Injury

• Kim SH
• Chu HJ
• Kang DH
• Song GA
• Cho M
• Yang US
• Kim HJ
• Chung HY

• KMID: 648675
• DOI: http://doi.org/10.3346/jkms.2002.17.2.193

Abstract

The involvement of NF-kappaB binding activity is known to be important in the mechanism of acute liver injury and in the induction of cyclooxygenase (COX-2). This study was performed to evaluate NF-kappaB binding activity and the expression of COX-2 in chronic liver injury induced by carbon tetrachloride (betaCCI(4)). Liver tissues from Sprague - Dawley rats were collected at 1, 3, 5, and 7th week after intraperitoneal injection of 0.1 mL of betaCCI(4)/100 g body weight twice a week. Reactive oxy-gen species (ROS) were measured in the postmitochondrial fraction by dichlorofluorescein formation with a fluorescent probe. An electrophoretic mobility shift assay was performed for NF-kappaB binding activity. Western blot was performed to measure the level of COX-1, COX-2, p65, p50, and I B proteins. ROS and NF-kappaB activity increased during the CCl4-induced chronic liver injury. The expression of nuclear p65 protein and p50 protein increased compared with that of the control, while the cytoplasmic I B protein decreased as the inflammation persisted. The expression of COX-2 in betaCCI(4)-treated rat liver increased compared with that of the control. It could be suggested that ROS produced by betaCCI(4) treatment increased NF-kappaB binding activity and thereby COX-2 expression, and these might be implicated in the progress of chronic liver damage.

Keyword

NF-kappaB; Prostaglandin-Endoperoxide Synthase; Carbon Tetrachloride; Liver Failure, Chronic

MeSH Terms

Animals
Biological Transport
Carbon Tetrachloride/administration & dosage/*adverse effects
Carbon Tetrachloride Poisoning/*metabolism/pathology
Cell Nucleus/metabolism
Cyclooxygenase 1
Cyclooxygenase 2
Cytoplasm/metabolism
I-kappa B Proteins/biosynthesis
Isoenzymes/*biosynthesis
Liver/drug effects/*injuries/pathology
Membrane Proteins
NF-kappa B/antagonists & inhibitors/*metabolism
NF-kappa B p50 Subunit
Prostaglandin-Endoperoxide Synthases/*biosynthesis
Protein Binding
Rats
Rats, Sprague-Dawley
Reactive Oxygen Species
Transcription Factor RelA