Inhibition of glucagon-signaling and downstream actions by interleukin 1beta and tumor necrosis factor alpha in cultured primary rat hepatocytes.
In cultured rat hepatocytes, glucagon increased the phosphoenolpyruvate carboxykinase (PCK1) mRNA by increasing cellular cAMP concentrations. The proinflammatory cytokines rhIL1beta and rhTNF alpha impaired the increase both in cAMP and PCK1 mRNA. Glucose formation from glycogen stimulated by glucagon was also attenuated by the cytokines, very likely due to the attenuation of the cAMP increase. Treatment of hepatocytes with the phosphodiesterase inhibitor IBMX or the inhibitory G-protein (G i) inactivating compound pertussis toxin did not abolish the inhibition of the glucagon-stimulated increase in cAMP by the cytokines indicating that phosphodiesterase and G i were not involved. The activation of adenylate cyclase by forskolin enhanced cAMP and PCK1 mRNA. Again, rhIL1beta and rhTNF alpha attenuated the increase in PCK1 mRNA, however, not that in cAMP. The stimulation of PCK1 mRNA increase with the nonhydrolyzable cAMP analogue CPT-cAMP was inhibited by rhIL1beta and rhTNF alpha indicating interference independent of changes in cAMP levels. It is concluded that rhIL1beta and rhTNF alpha inhibited glucagon-stimulated signal transduction at the site of cAMP formation. In addition, glucagon-stimulated PCK1 mRNA was attenuated independent of cAMP formation very likely on the transcriptional and/or post-transcriptional level.
Christ
B
B
Institut für Biochemie und Molekulare Zellbiologie, Georg-August-Universität, Humboldtallee, Göttingen, Germany. bruno.christ@medizin.unihalle.de
D000818:Animals; D002478:Cells, Cultured; D005576:Colforsin / Q000494:pharmacology; D000242:Cyclic AMP / Q000378:metabolism / Q000494:pharmacology; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D005934:Glucagon / Q000378:metabolism* / Q000494:pharmacology; D005941:Glucokinase / Q000235:genetics / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D006003:Glycogen / Q000378:metabolism; D022781:Hepatocytes / Q000187:drug effects* / Q000201:enzymology / Q000378:metabolism*; D006801:Humans; D007328:Insulin / Q000494:pharmacology; D053583:Interleukin-1beta / Q000494:pharmacology*; D008297:Male; D010729:Phosphoenolpyruvate Carboxykinase (GTP) / Q000235:genetics / Q000378:metabolism; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D007328:Insulin; D053583:Interleukin-1beta; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D005576:Colforsin; D006003:Glycogen; D005934:Glucagon; D000242:Cyclic AMP; D005941:Glucokinase; D010729:Phosphoenolpyruvate Carboxykinase (GTP); D005947:Glucose