Medium change amplifies mitogen-activated protein kinase-mediated prostaglandin E2 synthesis in Swiss 3T3 fibroblasts.
In Swiss 3T3 fibroblasts, changing the culture medium prior to stimulation resulted in an augmentation of bradykinin-induced prostaglandin E2 synthesis. The augmentation depended on the duration of the exposure to the fresh medium, with a maximum effect at 1 h. Fetal calf serum in the fresh medium was essential for augmented prostaglandin E2 synthesis. The medium change slightly augmented the bradykinin-induced increase in intracellular free Ca2+ concentration and phosphoinositide hydrolysis with a different time course from that for prostaglandin E2 synthesis. 4',5,7-Trihydroxyisoflavone (genistein) and 3,4-dihydroxybenzylidene-malononitrile (tyrphostin 23), inhibitors of tyrosine kinases, and 2'-amino-3'-methoxyflavone (PD98059), an inhibitor of mitogen-activated protein kinase (MAPK) kinase, attenuated the increase in prostaglandin E2 synthesis. Bradykinin caused phosphorylation of cytosolic phospholipase A2 and p42/p44 MAPK, which was augmented by the medium change. From the results, it is concluded that activation of MAPK and cytosolic phospholipase A2 is involved in the augmentation of prostaglandin E2 synthesis produced by the medium change.
Department of Pharmaceutical Molecular Biology, Faculty of Pharmaceutical Sciences, Tohoku University, Sendai, Japan.
K
K
Nakatani
N
N
Nakahata
Y
Y
Hamada
S
S
Tsurufuji
Y
Y
Ohizumi
D016475:3T3 Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D000818:Animals; D001920:Bradykinin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000037:antagonists & inhibitors / Q000502:physiology*; D003470:Culture Media / Q000494:pharmacology; D003600:Cytosol / Q000187:drug effects / Q000201:enzymology; D015232:Dinoprostone / Q000096:biosynthesis*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D005419:Flavonoids / Q000494:pharmacology; D019833:Genistein / Q000494:pharmacology; D006868:Hydrolysis / Q000187:drug effects; D051379:Mice; D010716:Phosphatidylinositols / Q000378:metabolism; D010741:Phospholipases A / Q000187:drug effects / Q000378:metabolism; D054467:Phospholipases A2; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors; D020032:Tyrphostins / Q000494:pharmacology
D003470:Culture Media; D004791:Enzyme Inhibitors; D005419:Flavonoids; D010716:Phosphatidylinositols; D020032:Tyrphostins; D019833:Genistein; D011505:Protein-Tyrosine Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D010741:Phospholipases A; D054467:Phospholipases A2; D015232:Dinoprostone; C060641:tyrphostin A23; D001920:Bradykinin; C093973:2-(2-amino-3-methoxyphenyl)-4H-1-benzopyran-4-one; D002118:Calcium