Arachidonic acid metabolism by rat alveolar epithelial cells.
Arachidonic acid release and metabolism by stimulated cultures of rat type II alveolar epithelial cells (94 +/- 2% pure) were studied. As compared with unstimulated cultures, a marked increase in the release of [14C]arachidonic acid from prelabeled cells was observed when the cells were incubated with the calcium ionophore A23187. Radioimmunoassay of unlabeled cultures demonstrated significant increases in the production of prostaglandin E2 greater than 6-Keto-prostaglandin F1 alpha greater than prostaglandin F2 alpha greater than thromboxane B2 with A23187 stimulation. Reverse-phase high performance liquid chromatography of media from cells prelabeled with [14C]arachidonic acid confirmed the identities and relative amounts of these metabolites. As expected, the production of these cyclooxygenase products was inhibited by indomethacin. Stimulation with A23187 led to no increment in immunoreactive leukotriene C4 production, but yielded a statistically significant but quantitatively small increment in leukotriene B4 production; its production by small numbers of contaminating macrophages cannot be ruled out. Analysis by high performance liquid chromatography of media from prelabeled cells after 30 minutes stimulation revealed no peaks of radioactivity coeluting with the lipoxygenase products leukotriene B4, leukotriene C4, or 5-, 12-, or 15-hydroxy-6,8,11,14-eicosatetraenoic acid. The results indicate that rat alveolar epithelial cells have the capacity to release arachidonic acid and metabolize it to an array of cyclooxygenase products. However, after stimulation, little or no lipoxygenase products accumulated in media. Thus, the alveolar epithelium may be a source of bioactive eicosanoids with potentially important roles in pulmonary physiology and pathophysiology.
Chauncey
J B
JB
Department of Internal Medicine, University of Michigan, Ann Arbor.
Peters-Golden
M
M
Simon
R H
RH
D000818:Animals; D001095:Arachidonic Acids / Q000378:metabolism*; D000001:Calcimycin / Q000494:pharmacology; D002851:Chromatography, High Pressure Liquid; D004847:Epithelial Cells; D004848:Epithelium / Q000201:enzymology / Q000378:metabolism; D007213:Indomethacin / Q000494:pharmacology; D007975:Leukotriene B4 / Q000096:biosynthesis; D008084:Lipoxygenase / Q000378:metabolism; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism; D011453:Prostaglandins / Q000096:biosynthesis; D011650:Pulmonary Alveoli / Q000166:cytology / Q000201:enzymology / Q000378:metabolism*; D011863:Radioimmunoassay; D051381:Rats; D013189:SRS-A / Q000096:biosynthesis; D013929:Thromboxane B2 / Q000096:biosynthesis
D001095:Arachidonic Acids; D011453:Prostaglandins; D013189:SRS-A; D007975:Leukotriene B4; D000001:Calcimycin; D013929:Thromboxane B2; D008084:Lipoxygenase; D011451:Prostaglandin-Endoperoxide Synthases; D007213:Indomethacin