Utilization by human amniocytes for prostaglandin synthesis of [1-14C]arachidonate derived from 2-[1-14C]arachidonylphosphatidylcholine associated with human fetal pulmonary surfactant.
The phospholipids of human fetal pulmonary surfactant prepared from term amniotic fluid contained arachidonic acid and its utilization for prostaglandin synthesis by amnion cells has been investigated. Cells were incubated with surfactant labelled with L-alpha-1-palmitoyl-2-[1-14C]arachidonylphosphatidylcholine. The uptake of radioactivity into amniocyte phospholipids increased with time and with the concentration of surfactant and after 2 h of incubation at 37 degrees C, 63% of the incorporated radioactivity was recovered in phosphatidylethanolamine (PE) and phosphatidylinositol (PI). Similar results were obtained when amniocytes were incubated with liposomes prepared from lipid extracts of surfactant, but when cells were incubated with liposomes prepared from synthetic lipids the transfer of radioactivity to PE and PI was only 27%. Fetal surfactant contained platelet activating factor (PAF) but the addition of the antagonist hexanolamino-PAF did not affect either the uptake or intracellular redistribution of surfactant arachidonate by amniocytes, nor did the addition of PAF affect the results obtained with liposomes prepared with synthetic lipids. Cells preincubated with surfactant labelled with 2-[1-14]arachidonylPC released radioactive arachidonate and prostaglandin E2 when stimulated with calcium ionophore A23187 or with phorbol ester. These data demonstrate that surfactant provides a source of arachidonate that can be utilized by amnion cells for prostaglandin synthesis.
Newman
G E
GE
Nuffield Department of Clinical Biochemistry, Radcliffe Infirmary, Oxford, UK.
Phizackerley
P J
PJ
López Bernal
A
A
D000650:Amnion / Q000378:metabolism*; D000653:Amniotic Fluid / Q000737:chemistry; D016718:Arachidonic Acid / Q000378:metabolism*; D002250:Carbon Radioisotopes; D005227:Fatty Acids / Q000032:analysis; D005333:Fetus / Q000378:metabolism*; D006801:Humans; D008081:Liposomes / Q000138:chemical synthesis; D010713:Phosphatidylcholines / Q000737:chemistry* / Q000494:pharmacology; D010740:Phospholipases / Q000378:metabolism; D010743:Phospholipids / Q000096:biosynthesis; D010972:Platelet Activating Factor / Q000494:pharmacology; D011453:Prostaglandins / Q000096:biosynthesis*; D011663:Pulmonary Surfactants / Q000737:chemistry*
D002250:Carbon Radioisotopes; D005227:Fatty Acids; D008081:Liposomes; D010713:Phosphatidylcholines; D010743:Phospholipids; D010972:Platelet Activating Factor; D011453:Prostaglandins; D011663:Pulmonary Surfactants; D016718:Arachidonic Acid; D010740:Phospholipases