Intracellular actions of group IIA secreted phospholipase A2 and group IVA cytosolic phospholipase A2 contribute to arachidonic acid release and prostaglandin production in rat gastric mucosal cells and transfected human embryonic kidney cells.
Gastric epithelial cells liberate prostaglandin E(2) in response to cytokines as part of the process of healing of gastric lesions. Treatment of the rat gastric epithelial cell line RGM1 with transforming growth factor-alpha and interleukin-1beta leads to synergistic release of arachidonate and production of prostaglandin E(2). Results with highly specific and potent phospholipase A(2) inhibitors and with small interfering RNA show that cytosolic phospholipase A(2)-alpha and group IIA secreted phospholipase A(2) contribute to arachidonate release from cytokine-stimulated RGM1 cells. In the late phase of arachidonate release, group IIA secreted phospholipase A(2) is induced (detected at the mRNA and protein levels), and the action of cytosolic phospholipase A(2)-alpha is required for this induction. Results with RGM1 cells and group IIA secreted phospholipase A(2)-transfected HEK293 cells show that the group IIA phospholipase acts prior to externalization from the cells. RGM1 cells also express group XIIA secreted phospholipase A(2), but this enzyme is not regulated by cytokines nor does it contribute to arachidonate release. The other eight secreted phospholipases A(2) were not detected in RGM1 cells at the mRNA level. These results clearly show that cytosolic and group IIA secreted phospholipases A(2) work together to liberate arachidonate from RGM1 cell phospholipids in response to cytokines.
Ni
Zhanglin
Z
Departments of Chemistry and Biochemistry, University of Washington, Seattle, WA 98195, USA.
Okeley
Nicole M
NM
Smart
Brian P
BP
Gelb
Michael H
MH
D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism; D018938:Caco-2 Cells; D015232:Dinoprostone / Q000378:metabolism; D005753:Gastric Mucosa / Q000166:cytology* / Q000378:metabolism*; D054501:Group II Phospholipases A2; D054513:Group IV Phospholipases A2; D006801:Humans; D007375:Interleukin-1 / Q000378:metabolism; D007668:Kidney / Q000166:cytology* / Q000196:embryology; D010741:Phospholipases A / Q000378:metabolism / Q000502:physiology*; D054467:Phospholipases A2; D011453:Prostaglandins / Q000378:metabolism; D034622:RNA Interference; D051381:Rats; D014162:Transfection
D007375:Interleukin-1; D011453:Prostaglandins; D016718:Arachidonic Acid; D010741:Phospholipases A; D054501:Group II Phospholipases A2; D054513:Group IV Phospholipases A2; C516389:PLA2G4A protein, human; D054467:Phospholipases A2; C516391:Pla2g4a protein, rat; D015232:Dinoprostone