Effect of protein kinase C activating agents on respiratory glycoconjugate release from feline airways.
Abnormal regulation of airway glycoprotein secretion may underlie many respiratory diseases. Experimental activation of the protein kinase C (PKC) family of cytosolic enzymes has been shown to induce a secretory response in many tissues. To estimate the effect of PKC activation on airway secretion, alteration in the amount of radiolabeled respiratory glycoconjugate (RGC) released into culture media was determined following feline airway explant exposure to PKC activating agents. Exposure to two known activators of PKC, phorbol 12-myristate 13-acetate (PMA) and mezerein (MEZ), resulted in profound increases in respiratory glycoconjugate release over a seven day experimental period. The response evolved over several hours and was dose dependent. Maximal RGC release, 90% above control, occurred 2 days after exposure to either PMA or MEZ. Pharmacological inhibition of the PKC effect using two PKC inhibitors, 1-(5-isoquinolinylsulfonyl)-2-methylpiperazine and sphingosine, resulted in dose-dependent antagonism of the maximal PMA (10(-7) M)-stimulated RGC release, suggesting altered PKC activity was responsible for augmenting RGC release. Since altered arachidonic acid metabolism has been implicated in mediating some PKC effects, eicosanoids were assayed in airway explant supernatants following PMA exposure. Enhanced release of both cyclooxygenase and lipoxygenase pathway products was detected by radioimmunoassay. Cotreatment of explants with PMA and an inhibitor of oxidative arachidonic acid metabolism, nordihydroguaiaretic acid, blocked RGC release. These data demonstrate prolonged augmentation of respiratory glycoconjugate release from airway explants following exposure to PKC-activating agents.
Rieves
R D
RD
Department of Critical Care Medicine, National Institutes of Health, Bethesda, Maryland 20892.
Lundgren
J D
JD
Logun
C
C
Wu
T
T
Shelhamer
J H
JH
D000818:Animals; D002415:Cats; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004224:Diterpenes; D015777:Eicosanoids / Q000378:metabolism; D004789:Enzyme Activation; D006001:Glycoconjugates / Q000378:metabolism*; D007700:Kinetics; D008168:Lung / Q000187:drug effects / Q000378:metabolism*; D009637:Masoprocol / Q000494:pharmacology; D009924:Organ Culture Techniques; D011493:Protein Kinase C / Q000378:metabolism*; D013729:Terpenes / Q000494:pharmacology*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology* / Q000633:toxicity
D004224:Diterpenes; D015777:Eicosanoids; D006001:Glycoconjugates; D013729:Terpenes; C011309:mezerein; D009637:Masoprocol; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate