David M. Olson

Estradiol-17β and 2-hydroxyestradiol-17β-induced differential production of prostaglandins by cells dispersed from human intrauterine tissues at parturition

Abstract Prostaglandin (PGE, 6-keto PGF 1α ) output by cells dispersed from human amnion and decidua in the presence of increasing levels (0–5000 ng/ml) of estradiol-17β (E 2 ) or 2-hydroxyestradiol-17β (2-OH E 2 ) was studied in relation to parturition. Tissues were obtained from women at term either before (CS) or after (SL) spontaneous labor and vaginal delivery. In the absence of estrogens, the output of both PGs from amnion increased significantly with labor. No significant increase in decidua PG output occurred with labor. Neither estrogen influenced CS amnion PG output. However, both E 2 and 2-OH E 2 stimulated SL amnion PGE output (2-OH E 2 >E 2 ) while having no affect on 6-keto PGF 1α output. Only the highest dose of 2-OH E 2 stimulated PGE output in CS decidua, but both estrogens significantly inhibited 6-keto PGF 1α output in this tissue. In SL decidua only 2-OH E 2 significantly stimulated PGE, and neither estrogen affected 6-keto PGF 1α output. These results might suggest that estrogens modulate PG biosynthesis at the level of endoperoxide to primary PG conversion.

The effects of mechanical stretching on fetal rat lung cell prostacyclin production

A model system was used to determine the effect of stretch on prostacyclin (PGI) production by organotypic fetal rat lung cultures grown on gelatin foam in vitro, measured by RIA of 6-keto-PGF1 alpha (6KF) in the culture medium. The stretching apparatus was programmable for stretch of varying frequency and duration. The effective stimuli for PGI production were: continuous pulsatile stretch greater than intermittent pulsatile stretch greater than permanent stretch (p less than 0.05). The rate of PGI production was greatest in the first 15 min of pulsatile stretch and was associated with a 70% increase in cAMP production (p less than 0.05). When the effect of magnitude of stretch was compared (15% vs 28% extension), there was a significant increase with a maximum in the 28% stretch group double that of the 15% stretch group (p less than 0.01). PGI production in response to pulsatile stretching was inhibited by indomethacin but not by pretreatment with cortisol. These results suggest that the production of PGI by lung cells may be significantly affected by the frequency and magnitude of pulsatile stretching.

Response of fetal rat lung fibroblasts to elevated oxygen concentrations after liposome-mediated augmentation of antioxidant enzymes.

Cultured fetal rat lung fibroblasts are growth-inhibited and have increased lactate dehydrogenase release and prostaglandin synthesis in response to 50% and 95% oxygen exposure. Extended exposure to 50% oxygen, but not to 95% oxygen, was associated with tolerance to the cytotoxic effects of oxygen. Pretreatment of fibroblasts with liposome-encapsulated superoxide dismutase and catalase also conferred protection against the cytotoxic effects of 50% and 95% oxygen. Exogenous enhancement of intracellular superoxide dismutase and catalase specific activities did not attenuate the growth inhibition or increased prostaglandin synthesis associated with exposure to 50% or 95% oxygen. The growth inhibition could not be attributed to an autocrine prostaglandin effect, since inhibition of prostaglandin synthesis with 50 microM ibuprofen did not prevent O2-mediated inhibition of DNA synthesis.

Production of prostaglandins by fetal rat lung type II pneumonocytes and fibroblasts

The output of prostaglandins I2, E2, F2 alpha and 13,14-dihydro-15-keto-PGF2 alpha (PGFM) from third passage day 20 rat fetal fibroblasts and type II alveolar pneumonocytes was studied. In 2 h incubations, the output levels for each cell type were: PGI2 greater than PGE2 much greater than PGF2 alpha = PGFM when cells were incubated with Ca2+ ionophore A23187 (10 microM) or arachidonic acid (1 microgram/ml).

Arachidonic acid incorporation into lipids of term human amnion

There were no differences in the rate or amount of (1-14C)-labeled arachidonic acid incorporated into triacylglycerides, diacylglycerides, or any phospholipid species of freshly dispersed term human amnion cells obtained before or after labor. Both phosphatidylcholine and phosphatidylethanolamine incorporated 14C-arachidonic acid in proportion to their molar percent of total amnion phospholipids, but phosphatidylinositol incorporated three times as much 14C-arachidonic acid, suggesting either a rapid turnover in this specific phospholipid pool or a greater specificity for the transfer of arachidonoyl-coenzyme A to lysophosphatidylinositol. No or little competition of 14C-arachidonic acid incorporation into triacylglycerides or phospholipids occurred with palmitic acid, linoleic acid, or gamma-linolenic acid. However, dihomo-gamma-linolenic acid, eicosapentaenoic acid, and unlabeled arachidonic acid were effective inhibitors. We conclude that the term amnion has high acyl transferase activity, that no change in the basal activity of this enzyme occurs with the onset of labor, and that a specific acyl transferase exists for 20-carbon polyunsaturated fatty acids.

STIMULATION OF OVINE MYOMETRIAL ACTIVITY BY 6-KETO PROSTAGLANDIN-E1

6-keto prostaglandin E1 (6KE) is a metabolite of PGI2, which we have shown previously inhibits spontaneous myometrial activity. In the present study we examined the effects of 6KE on uterine electrical and mechanical activity in non-pregnant ovariectomized sheep. 6KE stimulated uterine activity in a dose-dependent fashion. The effect was enhanced by pre-treatment with estradiol (E2). It was not influenced by pre-treatment with meclofenamic acid and was not associated with significant changes in the concentrations of 13,14 dihydro 15-keto PGF2 alpha in vena cava plasma. After E2 treatment, 6KE had 0.2-0.3 of the stimulatory activity of PGF2 alpha. In the absence of E2, the uterine response to both 6KE and PGF2 alpha was decreased. In animals in which spontaneous myometrial activity was inhibited by PGI2, the uterus remained responsive to 6KE. We conclude that in the ovariectomized non-pregnant sheep 6KE stimulates uterine activity, and that the effect is independent of endogenous PG production.