Regulation of corticotropin-releasing factor receptor type 2beta messenger ribonucleic acid by interleukin-1beta in rat vascular smooth muscle cells.
OBJECTIVE
Corticotropin-releasing factor receptor type 2 (CRF R2) messenger RNA (mRNA) expression in the rodent heart or vessels is modulated by exposure to urocortin and glucocorticoids. In addition, we previously found that incubation with a variety of cytokines, such as interleukin (IL)-1 and IL-6, and tumor necrosis factor-alpha also reduced CRF R2beta mRNA expression, with IL-1beta being the most effective. In this study, we further explored the regulation of CRF R2beta mRNA levels by IL-1beta in the rat vascular smooth muscle A7r5 cells.


METHODS
A7r5 cells were incubated with IL-1beta, urocortin, or both for 6 h, after pre-incubating with or without anti-IL-1beta antibody (Ab) for 30 min, and then CRF R2beta mRNA levels were measured by RNase protection assay. Cells were incubated with lipopolysaccharides, IL-1beta, IL-6, dexamethasone, forskolin, or urocortin for 20 min, and then intracellular cAMP was measured by cAMP RIA.


RESULTS
IL-1beta produced a significant time-dependent decrease in CRF R2beta mRNA levels. Combined urocortin and IL-1beta administration did not have synergistic effects on the decrease in CRF R2beta mRNA levels. IL-1beta Ab failed to block the ability of urocortin to regulate CRF R2beta mRNA levels, suggesting that urocortin regulated CRF R2beta mRNA levels via another pathway than IL-1beta production. Urocortin induced the intracellular cAMP production in A7r5 cells, while IL-1beta failed to induce it.


CONCLUSIONS
The multifactorial regulation of CRF R2beta mRNA expression in the A7r5 cells serves to limit the inotropic and chronotropic effects of CRF R2 agonists such as urocortin during prolonged physical or immune challenge.
Kageyama
K
K
Third Department of Internal Medicine, Hirosaki University School of Medicine, Hirosaki, Japan. kkageyama@hkg.odn.ne.jp
Suda
T
T
D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D000906:Antibodies / Q000494:pharmacology; D002478:Cells, Cultured; D003346:Corticotropin-Releasing Hormone / Q000276:immunology* / Q000378:metabolism / Q000494:pharmacology; D000242:Cyclic AMP / Q000276:immunology / Q000378:metabolism; D016207:Cytokines / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology; D003907:Dexamethasone / Q000494:pharmacology; D015536:Down-Regulation / Q000187:drug effects / Q000276:immunology*; D004347:Drug Interactions / Q000502:physiology; D005786:Gene Expression Regulation / Q000187:drug effects / Q000276:immunology; D007375:Interleukin-1 / Q000037:antagonists & inhibitors / Q000276:immunology* / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D012333:RNA, Messenger / Q000187:drug effects / Q000378:metabolism*; D051381:Rats; D018019:Receptors, Corticotropin-Releasing Hormone / Q000235:genetics*; D013312:Stress, Physiological / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D054832:Urocortins
D000893:Anti-Inflammatory Agents; D000906:Antibodies; C101531:CRF receptor type 2; D016207:Cytokines; D007375:Interleukin-1; D012333:RNA, Messenger; D018019:Receptors, Corticotropin-Releasing Hormone; D054832:Urocortins; D003907:Dexamethasone; D003346:Corticotropin-Releasing Hormone; D000242:Cyclic AMP