Uptake of the phospholipase A2 inhibitor 1-dodecyl 2-[1-14C] octanamido-sn-2-deoxy glycero-3-phosphocholine by peritoneal macrophages.
The [14C] phospholipid analogue 1-dodecyl-2-[1-14C] octanamido-sn-2-deoxy glycero-3-phosphocholine was synthetized. With 2 short fatty chains linked by alkyl and amido bonds to positions 1 and 2 of the glycerophosphate backbone, it was an inhibitor of phospholipase A2 in ionophore A23187-stimulated macrophages. Its uptake by rat peritoneal macrophages and its resistance towards phospholipases A2 were determined at nanomolar or micromolar concentrations in the culture medium. A control substrate for phospholipases A2 activity was established with the lecithin 1-octadecanoyl 2-[3H] eicosatetraenoyl-sn-glycero-3-phosphocholine ([3H] 20:4-GPC), a source of [3H] arachidonic acid after cleavage at position 2. Non-stimulated- or ionophore A23187-stimulated macrophages incorporated extensively the [14C] phospholipid analogue added at 30-4000 nM. At 4000 nM which induced 50% inhibition of the phospholipase, 40% of the dose was found associated with the [14C] phospholipids of 2 x 10(6) stimulated macrophages after 120 min incubation, while only low amounts of [14C] non-phosphorous lipids were detected. It is concluded that the [14C] phospholipid analogue was readily taken up by the macrophages with limited hydrolysis.
Bourass
J
J
Laboratoire de synthèse et d'étude des substances naturelles à activité biologique, pôle sciences, La Rochelle, France.
Boucrot
P
P
Letourneux
Y
Y
Gandemer
G
G
Petit
J Y
JY
D000818:Animals; D002250:Carbon Radioisotopes; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000378:metabolism*; D050356:Lipid Metabolism; D008055:Lipids / Q000032:analysis; D017737:Macrophages, Peritoneal / Q000378:metabolism*; D008297:Male; D009994:Osmolar Concentration; D010741:Phospholipases A / Q000037:antagonists & inhibitors* / Q000138:chemical synthesis / Q000378:metabolism*; D054467:Phospholipases A2; D010742:Phospholipid Ethers / Q000138:chemical synthesis / Q000378:metabolism*; D051381:Rats; D017208:Rats, Wistar; D014316:Tritium
C109624:1-dodecyl-2-octanamido-sn-2-deoxyglycero-3-phosphocholine; D002250:Carbon Radioisotopes; D004791:Enzyme Inhibitors; D008055:Lipids; D010742:Phospholipid Ethers; D014316:Tritium; D010741:Phospholipases A; D054467:Phospholipases A2