Prostaglandin E2 production by endotoxin-stimulated alveolar macrophages is regulated by phospholipase C pathways.
BACKGROUND
Eicosanoids play an important role in many aspects of systemic inflammatory responses and host defense. Although the synthesis of eicosanoids by different enzymes has been elucidated, the regulatory mechanism of eicosanoid production is not clear. We designed this study to investigate the hypothesis that PGE2 production by endotoxin (lipopolysaccharide; LPS)-stimulated macrophages (MO) is dependent on phospholipase C (PLC) signaling pathways.


METHODS
Rabbit alveolar macrophages (MO) were obtained by bronchoalveolar lavage. MO were suspended in RPMI-1640 medium at 1 x 10(6)/mL and were exposed to Escherichia coli LPS (10 ng/mL) +/- various agonists and antagonists of PLC and its secondary messengers. After 24 hours of incubation, prostaglandin E2 (PGE2) production was measured by ELISA.


RESULTS
LPS-activated MO produced four times as much PGE2 as did control unstimulated MO. The increase in PGE2 production was inhibited by PLC inhibitors (U73122 or D609) and a low-molecular-weight PLA2 inhibitor, manoalide. An increase in intracellular calcium and activation of both the calmodulin and protein kinase C kinase pathways increase PGE2 production.


CONCLUSIONS
PGE2 production is intimately dependent on several phospholipases. Production is not only dependent on low-molecular-weight PLA2 cleavage of arachidonic acid from membrane phospholipids, but also by-products of PLC activation. PLC-dependent intracellular Ca-calmodulin signaling and protein kinase C activation provide significant modulation of PGE2 production.
Lo
C J
CJ
Department of Surgery, University of California, Los Angeles 90024, USA.
Cryer
H G
HG
Maier
R V
RV
D000818:Animals; D015232:Dinoprostone / Q000096:biosynthesis*; D008070:Lipopolysaccharides; D016676:Macrophages, Alveolar / Q000378:metabolism*; D008297:Male; D009102:Multiple Organ Failure / Q000503:physiopathology; D011817:Rabbits; D015398:Signal Transduction / Q000502:physiology*; D010738:Type C Phospholipases / Q000502:physiology*
D008070:Lipopolysaccharides; D010738:Type C Phospholipases; D015232:Dinoprostone