The protective effect of protein kinase C and adenosine triphosphate-sensitive potassium channel agonists against inflammation in rat endothelium and vascular smooth muscle in vitro and in vivo.
Volatile anesthetic pretreatment protects the vasculature from inflammation-induced injury via mechanisms involving the activation of adenosine triphosphate-sensitive potassium (K(ATP)) channels and/or protein kinase C (PKC). Therefore, we hypothesized that K(ATP) and PKC agonists may mimic the protective effects of volatile anesthetics in vitro and in vivo. In vitro, rat vascular smooth muscle cells (VSM) and aortic endothelial cells (AEC) were used to evaluate whether pretreatment with a K(ATP) agonist, cromakalim (CRK), or a PKC agonist, phorbol 12-myristate 13-acetate (PMA), decreases lipopolysaccharide (LPS)-induced cell injury. Cell survival was determined by trypan blue staining after 6 h. In vivo, rats received systemic LPS or saline with or without pretreatment with PMA or CRK. Mean arterial blood pressure, the response to endothelium-dependent (acetylcholine; ACH) and -independent (sodium nitroprusside) vasodilators, and arterial blood gases were determined after 6 h. Cell survival in VSM and AEC control cultures was more than 90%, which was not altered in the presence of PMA or CRK, whereas LPS significantly decreased cell survival. PMA (0.1-10 microM) significantly attenuated the LPS-induced decrease in cell survival by 28%-37% in VSM and 39%-53% in AEC, and CRK (1 mM) increased cell survival by 24% in VSM and 22% in AEC. In vivo, PMA and CRK pretreatment had no significant effect on measured variables in control rats. LPS decreased mean arterial blood pressure and vasodilation to ACH and sodium nitroprusside and caused hypoglycemia. PMA, but not CRK, increased ACH-dependent vasodilation (46%) at 6 h, but neither agonist altered the other detrimental effects of LPS. In conclusion, PKC and K(ATP) agonists appear to protect AEC and VSM cells against inflammation in vitro, but the systemic administration of PKC and K(ATP) agonists appeared to exert minimal or no protection in our in vivo model.
Department of Anesthesiology, PO Box 800710, University of Virginia Health System, Charlottesville, VA 22908-0710, USA.
Roman V
RV
Plachinta
Manuela J M
MJ
de Klaver
John K
JK
Hayes
George F
GF
Rich
D018528:ATP-Binding Cassette Transporters; D000818:Animals; D000893:Anti-Inflammatory Agents; D001784:Blood Gas Analysis; D001794:Blood Pressure / Q000187:drug effects; D002452:Cell Count; D016923:Cell Death / Q000187:drug effects; D019806:Cromakalim / Q000494:pharmacology; D004730:Endothelium, Vascular / Q000473:pathology*; D020536:Enzyme Activators / Q000494:pharmacology*; D054086:KATP Channels; D008070:Lipopolysaccharides / Q000633:toxicity; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000473:pathology*; D015221:Potassium Channels / Q000819:agonists*; D024661:Potassium Channels, Inwardly Rectifying; D011493:Protein Kinase C / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014657:Vasculitis / Q000473:pathology / Q000517:prevention & control*; D014665:Vasodilator Agents / Q000494:pharmacology
D018528:ATP-Binding Cassette Transporters; D000893:Anti-Inflammatory Agents; D020536:Enzyme Activators; D054086:KATP Channels; D008070:Lipopolysaccharides; D015221:Potassium Channels; D024661:Potassium Channels, Inwardly Rectifying; D014665:Vasodilator Agents; C092520:uK-ATP-1 potassium channel; D019806:Cromakalim; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate