Effects of mitogen-activated protein kinase inhibitors on tumor necrosis factor-alpha gene expression and apoptosis induction in cultured human fetal membrane chorion cells infected with influenza virus.
OBJECTIVE
We investigated the involvement of p38 mitogen-activated protein (MAP) kinase in tumor necrosis factor (TNF)-alpha gene expression, apoptosis induction and virus replication in cultured human fetal membrane chorion cells infected with influenza virus.


METHODS
Influenza virus-infected chorion cells were incubated in the absence or presence of inhibitors of p38 MAP kinase, SB203580 and SB202190. TNF-alpha mRNA and hemagglutinin viral RNA (HA vRNA) were amplified with reverse transcriptase-polymerase chain reaction techniques. TNF-alpha protein concentrations were determined by enzyme-liked immunosorbent assay. The extent of apoptosis induction was estimated by DNA agarose gel electrophoresis. Pyrrolidine dithiocarbamate (PDTC) and ribavirin, which have been shown to inhibit apoptosis induction via the inhibition of viral gene replication, were used as positive control reagents.


RESULTS
PDTC and ribavirin inhibited the accumulation of TNF-alpha mRNA and HA vRNA in the virus-infected chorion cells, resulting in the suppression of TNF-alpha protein secretion. Both SB203580 and SB202190 suppressed TNF-alpha protein secretion, but not the accumulation of TNF-alpha mRNA as well as HA vRNA and the induction of apoptosis.


CONCLUSIONS
These results suggest that p38 MAP kinase pathway is critical in TNF-alpha gene expression at a post-transcriptional level but not in the apoptosis induction and influenza virus replication in cultured chorion cells.
Uchide
Noboru
N
Department of Clinical Molecular Genetics, School of Pharmacy, Tokyo University of Pharmacy and Life Science, Tokyo, Japan. uchide@ps.toyaku.ac.jp
Ohyama
Kunio
K
Bessho
Toshio
T
Toyoda
Hiroo
H
D000998:Antiviral Agents / Q000494:pharmacology; D017209:Apoptosis; D002478:Cells, Cultured; D002823:Chorion / Q000166:cytology / Q000187:drug effects / Q000821:virology*; D053938:DNA Fragmentation; D004587:Electrophoresis, Agar Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005333:Fetus; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D053118:Influenza A Virus, H1N1 Subtype / Q000276:immunology / Q000502:physiology*; D008954:Models, Biological; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D011725:Pyridines / Q000494:pharmacology; D011759:Pyrrolidines / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics; D012367:RNA, Viral / Q000032:analysis / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012254:Ribavirin / Q000494:pharmacology; D013859:Thiocarbamates / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics*; D048051:p38 Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors*
D000998:Antiviral Agents; D007093:Imidazoles; D047428:Protein Kinase Inhibitors; D011725:Pyridines; D011759:Pyrrolidines; D012333:RNA, Messenger; D012367:RNA, Viral; D013859:Thiocarbamates; D014409:Tumor Necrosis Factor-alpha; C020972:pyrrolidine dithiocarbamic acid; D012254:Ribavirin; D048051:p38 Mitogen-Activated Protein Kinases; C093642:SB 203580