Lipopolysaccharide induces time-dependent increases in prostaglandin H synthase-2 and cytosolic phospholipase A2 mRNA in cultured human microvessel-derived endothelial cells.
The effect of lipopolysaccharide (LPS) upon the cellular content of mRNA for several enzymes associated with the arachidonic acid cascade was determined in human microvessel-derived endothelial cells. Cells were treated with either vehicle or 10 ng/mL LPS for up to 24 h. Reverse transcription followed by DNA amplification and Southern blotting were used to quantify mRNA for prostaglandin H synthase-1, prostaglandin H synthase-2, cytoplasmic PLA2, and a group II secretory PLA2. LPS treatment resulted in an increase in prostaglandin H synthase-2 mRNA after 4 h with a second peak occurring at 12 h of incubation. The expression of prostaglandin H synthase-1 mRNA was not altered by LPS treatment. An increase in cytoplasmic PLA2 mRNA but not group II PLA2 mRNA was seen after 12 h. These data demonstrate that LPS can increase mRNA production for the inducible form of prostaglandin H synthase and for cytoplasmic PLA2 in a time-dependent manner in human microvessel-derived endothelial cells. These multiple, specific increases in the prostaglandin H synthase-2 and phospholipase A2 mRNA values suggest a complex interaction between bacterial LPS and the endothelial cell eicosanoid system.
Flynn
J T
JT
Department of Physiology, Thomas Jefferson University, Philadelphia, Pennsylvania 19107, USA.
Hoff
H
H
D001483:Base Sequence; D015139:Blotting, Southern; D002478:Cells, Cultured; D003600:Cytosol / Q000187:drug effects / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D008070:Lipopolysaccharides / Q000633:toxicity*; D008969:Molecular Sequence Data; D010741:Phospholipases A / Q000235:genetics / Q000378:metabolism*; D054467:Phospholipases A2; D044262:Prostaglandin H2; D011463:Prostaglandins H / Q000235:genetics / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism*; D018805:Sepsis / Q000378:metabolism* / Q000473:pathology; D014158:Transcription, Genetic
D008070:Lipopolysaccharides; D011463:Prostaglandins H; D012333:RNA, Messenger; D044262:Prostaglandin H2; D010741:Phospholipases A; D054467:Phospholipases A2