Effects of prostacyclin analogs on the synthesis of tissue factor, tumor necrosis factor-alpha and interleukin-1 beta in human monocytic THP-1 cells.
Previous studies have shown that prostacyclin analogs can inhibit the expression of tissue factor (TF) procoagulant activity by human monocytes. The present studies have investigated this phenomenon further, by using a plasma coagulation assay to measure cellular TF activity, an immunoassay to measure TF antigen and reverse transcription/polymerase chain reaction with appropriate oligomer primers to measure TF mRNA. Iloprost and cicaprost inhibited lipopolysaccharide-induced increases in TF activity, antigen and mRNA (50% inhibition, 2-8 nM), with no apparent effect on TF mRNA stability. These agents therefore act at or before the level of transcription of the TF gene. The analogs were more potent inhibitors of tumor necrosis factor-alpha synthesis (50% inhibition at 334 +/- 40 pM cicaprost or 846 +/- 182 pM iloprost) and extraordinarily potent when combined with a phosphodiesterase inhibitor (50% inhibition at 101 +/- 31 pM iloprost in the presence of 20 microM isobutylmethylxanthine). Iloprost and cicaprost were less potent in inhibiting the synthesis of interleukin-1 beta (50% inhibition, 50-100 nM). Cicaprost inhibited lipopolysaccharide-induced increases in mRNA levels for TF, tumor necrosis factor-alpha and interleukin-1 beta; differential potency was again observed. We conclude that these three important monocyte functions can be down-regulated by prostacyclin analogs, and with differential sensitivity. Furthermore, the extreme sensitivity of tumor necrosis factor-alpha synthesis to inhibition suggests that such inhibition may be a major physiological function of prostacyclin itself.
Crutchley
D J
DJ
Miami Heart Research Institute, Miami Beach, Florida.
Conanan
L B
LB
Que
B G
BG
D015056:1-Methyl-3-isobutylxanthine / Q000494:pharmacology; D001483:Base Sequence; D000242:Cyclic AMP / Q000032:analysis; D011464:Epoprostenol / Q000031:analogs & derivatives* / Q000494:pharmacology; D006801:Humans; D016285:Iloprost / Q000494:pharmacology*; D007375:Interleukin-1 / Q000096:biosynthesis* / Q000235:genetics; D008969:Molecular Sequence Data; D009000:Monocytes / Q000187:drug effects*; D012333:RNA, Messenger / Q000032:analysis; D013925:Thromboplastin / Q000096:biosynthesis* / Q000235:genetics; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis* / Q000235:genetics
D007375:Interleukin-1; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D013925:Thromboplastin; D011464:Epoprostenol; D000242:Cyclic AMP; D016285:Iloprost; C043867:cicaprost; D015056:1-Methyl-3-isobutylxanthine