PMID- 15924871 OWN - NLM STAT- MEDLINE DCOM- 20050714 LR - 20161124 IS - 0014-4800 (Print) IS - 0014-4800 (Linking) VI - 78 IP - 3 DP - 2005 Jun TI - The p105/50 NF-kappaB pathway is essential for Mallory body formation. PG - 198-206 AB - To determine if nuclear factor-kappaB (NF-kB) plays a role in Mallory body (MB) formation, quantitative real-time RT-PCR assay was used to measure liver NF-kappaB1/p105 mRNA levels in 4 different groups of mice. Group 1: mice given IP saline for 15 weeks; group 2: mice fed diethyl 1,4-dihydro-2,4,6,-trimethyl-3,5-pyridinedicarboxylate (DDC) for 10 weeks when MBs were formed; group3: mice fed DDC 10 weeks, then withdrawn 5 weeks when MBs disappeared; group 4: mice fed DDC 10 weeks, withdrawn 4 weeks, then fed DDC+chlormethiazole (CMZ) for 1 week when MBs again formed. The mRNA for p105 NF-kappaB expression was significantly increased in the livers of mice treated with DDC (group 2) and DDC+CMZ (group 4) compared with the control livers (group 1) as well as the drug-withdrawal livers (group 3). Primary cultures of hepatocytes from drug-primed mice (the group 4 mice were withdrawn for another 4 weeks when the MBs had disappeared) were studied. The hepatocytes from drug-primed mice were MB free when isolated and used for primary culture. MBs began to form spontaneously within their cytoplasm after 2-3 days of culture. The NF-kappaB inhibitor (NF-kappaBi), a cell-permeable quinazoline compound that acts as a potent inhibitor of NF-kappaB transcriptional activation, was added to the medium 3 h after planting the cultures of liver cells. No MBs formed in the cells treated with 10 microM, 1 microM, and 0.1 microM NF-kappaBi for 6 days. MBs still formed in the cells treated with 10 nM NF-kappaBi for 6 days. Both DDC-primed and normal control liver cells began to enlarge and elongate after a few hours of culture. In contrast, the cells treated with NF-kappaBi stayed polyhedral in shape just as they appeared prior to culturing. The level of NF-kappaB1/p105 mRNA significantly increased in DDC-primed hepatocytes after 24 h of culture and in normal control hepatocytes after 48 h of culture. In DDC-primed hepatocytes, NF-kappaBi 0.1 muM treatment for 6 days significantly decreased mRNA expression of Src, p105/NF-kappaB1, ERK1, MEKK1, and JNK1/2. In normal control liver cells, NF-kappaBi treatment decreased mRNA expression of Src and JNK1 and stimulated the mRNA expression of p105/NF-kappaB1 and Junk2. NF-kappaBi treatment significantly decreased the total ERK1/2 protein and further decreased the phosphorylated (activated) form of ERK1/2 in the cultured hepatocytes. The results indicate that the p105 NF-kappaB pathway which putatively regulates ERK at both the transcriptional and post-translational levels regulates MB formation by way of changes in gene expression. FAU - Nan, Li AU - Nan L AD - Department of Pathology, Harbor-UCLA Medical Center, 1000 W. Carson Street, Torrance, CA 90502, USA. FAU - Wu, Yong AU - Wu Y FAU - Bardag-Gorce, Fawzia AU - Bardag-Gorce F FAU - Li, Jun AU - Li J FAU - French, Barbaba A AU - French BA FAU - Wilson, La Toyia AU - Wilson LT FAU - French, Samuel W AU - French SW LA - eng GR - NIH/NIAAA 011999/AA/NIAAA NIH HHS/United States GR - NIH/NIAAA08116/AA/NIAAA NIH HHS/United States PT - Comparative Study PT - Journal Article PT - Research Support, N.I.H., Extramural PT - Research Support, U.S. Gov't, P.H.S. DEP - 20050209 PL - Netherlands TA - Exp Mol Pathol JT - Experimental and molecular pathology JID - 0370711 RN - 0 (Antigens, Nuclear) RN - 0 (Chromosomal Proteins, Non-Histone) RN - 0 (Dihydropyridines) RN - 0 (Enzyme Inhibitors) RN - 0 (NF-kappa B) RN - 0 (NF-kappa B p50 Subunit) RN - 0 (Protein Precursors) RN - 0 (Quinazolines) RN - 0 (RNA, Messenger) RN - 0C5DBZ19HV (Chlormethiazole) RN - 1150-55-6 (diethyl 1,4-dihydro-2,4,6-trimethyl-3,5-pyridinedicarboxylate) RN - EC 2.7.11.24 (JNK Mitogen-Activated Protein Kinases) RN - EC 2.7.11.24 (Mitogen-Activated Protein Kinase 3) RN - EC 2.7.11.25 (MAP Kinase Kinase Kinase 1) RN - EC 2.7.12.2 (MAP Kinase Kinase 4) RN - EC 2.7.12.2 (Mitogen-Activated Protein Kinase Kinases) SB - IM MH - Animals MH - Antigens, Nuclear/drug effects/metabolism MH - Blotting, Western MH - Cells, Cultured MH - Chemical and Drug Induced Liver Injury MH - Chlormethiazole/pharmacology MH - Chromosomal Proteins, Non-Histone/drug effects/metabolism MH - Dihydropyridines/toxicity MH - Enzyme Inhibitors/pharmacology MH - Hepatocytes/drug effects/metabolism/pathology MH - Inclusion Bodies/*metabolism/pathology MH - JNK Mitogen-Activated Protein Kinases/metabolism MH - Liver/drug effects/metabolism/*pathology MH - Liver Diseases/metabolism/*pathology MH - MAP Kinase Kinase 4 MH - MAP Kinase Kinase Kinase 1/metabolism MH - Male MH - Mice MH - Mitogen-Activated Protein Kinase 3/metabolism MH - Mitogen-Activated Protein Kinase Kinases/metabolism MH - *Models, Biological MH - NF-kappa B/drug effects/*metabolism MH - NF-kappa B p50 Subunit MH - Protein Precursors/drug effects/*metabolism MH - Quinazolines/pharmacology MH - RNA, Messenger/analysis MH - Reverse Transcriptase Polymerase Chain Reaction MH - Signal Transduction/physiology EDAT- 2005/06/01 09:00 MHDA- 2005/07/15 09:00 CRDT- 2005/06/01 09:00 PHST- 2004/11/15 00:00 [received] PHST- 2004/12/03 00:00 [accepted] PHST- 2005/06/01 09:00 [pubmed] PHST- 2005/07/15 09:00 [medline] PHST- 2005/06/01 09:00 [entrez] AID - S0014-4800(04)00127-3 [pii] AID - 10.1016/j.yexmp.2004.12.002 [doi] PST - ppublish SO - Exp Mol Pathol. 2005 Jun;78(3):198-206. doi: 10.1016/j.yexmp.2004.12.002. Epub 2005 Feb 9.