Cytokine mRNA decay is accelerated by an inhibitor of p38-mitogen-activated protein kinase.
OBJECTIVE
To identify the site(s) in tumor necrosis factor (TNFalpha), interleukin-6 (IL-6), and macrophage inflammatory protein-1alpha (MIP-1alpha) biosynthesis that is blocked by SB202190, a selective inhibitor of p38-mitogen activated protein kinase (p38).


METHODS
Human blood monocytes isolated by centrifugal elutriation.


METHODS
Monocytes were stimulated with lipopolysaccharide in the presence of 0, 0.3, 1 and 3 microM SB202190. Induced TNFalpha, IL-6, and MIP-1alpha protein and mRNA were measured by ELISA and quantitative RT-PCR, respectively. The half-lives of cytokine mRNA levels were determined following treatment of cells with actinomycin D or SB202190.


RESULTS
SB202190 suppressed >60% of lipopolysaccharide-induced TNFalpha, IL-6, and MIP-1alpha protein and mRNA expression. Suppressed mRNA levels could be attributed to a >2 to 7-fold reduction in cytokine mRNA half-lives. In contrast, SB202190 did not destabilize mRNAs encoding interferon-induced gene 15 protein and glyceraldehyde-3-phosphate dehydrogenase.


CONCLUSIONS
Specific mRNA destabilization represents an important and novel site of action for the cytokine suppressive effects of p38 inhibitors.
Wang
S W
SW
Department of Biology, Amgen Inc., Boulder, CO 80301, USA.
Pawlowski
J
J
Wathen
S T
ST
Kinney
S D
SD
Lichenstein
H S
HS
Manthey
C L
CL
D002478:Cells, Cultured; D054405:Chemokine CCL3; D054407:Chemokine CCL4; D016207:Cytokines / Q000097:blood / Q000235:genetics*; D004791:Enzyme Inhibitors / Q000097:blood / Q000494:pharmacology; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007093:Imidazoles / Q000097:blood / Q000494:pharmacology; D015850:Interleukin-6 / Q000096:biosynthesis / Q000097:blood / Q000235:genetics; D007700:Kinetics; D008070:Lipopolysaccharides / Q000494:pharmacology; D019402:Macrophage Inflammatory Proteins / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000097:blood / Q000235:genetics; D020928:Mitogen-Activated Protein Kinases / Q000037:antagonists & inhibitors* / Q000097:blood; D009000:Monocytes / Q000187:drug effects / Q000378:metabolism; D011725:Pyridines / Q000097:blood / Q000494:pharmacology; D020871:RNA Stability / Q000187:drug effects; D012333:RNA, Messenger / Q000097:blood / Q000737:chemistry / Q000187:drug effects*; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics; D048051:p38 Mitogen-Activated Protein Kinases; D000093242:Interleukin-6 Inhibitors
D054405:Chemokine CCL3; D054407:Chemokine CCL4; D016207:Cytokines; D004791:Enzyme Inhibitors; D007093:Imidazoles; D015850:Interleukin-6; D008070:Lipopolysaccharides; D019402:Macrophage Inflammatory Proteins; D020928:Mitogen-Activated Protein Kinases; D011725:Pyridines; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D048051:p38 Mitogen-Activated Protein Kinases; D000093242:Interleukin-6 Inhibitors; C090942:4-(4-fluorophenyl)-2-(4-hydroxyphenyl)-5-(4-pyridyl)imidazole