L-arginine prevents hypoxia-induced vasoconstriction in dual-perfused human placental cotyledons.

INTRODUCTION: Chronic hypoxia in the uteroplacental unit is associated with increased resistance to blood flow in the fetal-placental circulation. These changes can lead to adverse cardiovascular events in adulthood. This study investigates whether L-arginine (substrate for nitric oxide synthase (NOS) or endothelin-A receptor antagonist BQ123 administration reverses hypoxia-induced changes in perfusion pressure in the fetal compartment in dual-perfused placental cotyledons. METHODS: Human placental cotyledons (n = 15) from term deliveries (n = 15) were perfused with Krebs solution from maternal and fetal sides. Normal and reduced oxygen tension conditions were sequentially created in the perfused maternal compartment. Fetal perfusion pressure was continuously monitored. 1 mM L-arginine, D-arginine (an enantiomer of L-arginine and not a substrate for NOS), and BQ123 or normal saline were administered to the fetal compartment; L-arginine was also administered to the maternal compartment prior to maternal side hypoxia. Changes in perfusion pressure were compared between groups. RESULTS: Maternal hypoxia increased (19 ± 6%) perfusion pressure and this was blunted by L-arginine injection (3 ± 5%; p = 0.006) into the fetal compartment. L-arginine in the maternal compartment had no significant effect (22 ± 4% with L-arginine vs.14 ± 3% at control) on perfusion pressure. Similarly, D-arginine (23 ± 11% vs.19 ± 8% at control) or BQ123 (12 ± 3% vs.13 ± 3% at control) in the fetal compartment did not blunt the hypoxia-induced increase in perfusion pressure. CONCLUSIONS: Fetal vasoconstriction induced by maternal hypoxia is blunted by NO synthase substrate L-arginine, but not by D-arginine, in the fetal compartment, suggesting the involvement of NO synthesis in regulating the hypoxia-induced fetal vasoconstriction. Endothelin A receptor-related mechanisms does not appear to play a role in the maternal hypoxia-induced fetal vasoconstriction.

Challenge with ovalbumin antigen increases uterine and cervical contractile activity in sensitized guinea pigs.

OBJECTIVE: The aims of this study were to investigate the effects of ovalbumin challenge on uterine and cervical contractility, intrauterine pressure, and uterine electromyography activity in sensitized guinea pigs. STUDY DESIGN: Guinea pigs were sensitized by injection of ovalbumin-aluminum hydroxide suspension. Control animals were injected with the aluminum hydroxide suspension only. On days 55-57 of pregnancy, longitudinal uterine and cervical strips from guinea pigs were prepared for isometric tension recording. Nonpregnant guinea pigs were outfitted with telemetric transducers to record intrauterine pressure and uterine electromyography. RESULTS: Ovalbumin significantly increased contractility of uterine and cervical strips from sensitized versus nonsensitized animals. These effects were abolished by histamine H(1) receptor antagonist in uterine strips and by histamine H(1) receptor antagonist and a mast cell stabilizer in cervical strips from sensitized animals. Cyclooxygenase and 5-lipoxygenase inhibitors had no significant effect on the response to ovalbumin. Treatment with ovalbumin in vivo significantly increased intrauterine pressure and uterine electromyography activity in sensitized but not in nonsensitized, animals. CONCLUSION: Our findings indicate that type I hypersensitivity reactions may be important in mediating uterine contractility in pregnant and nonpregnant states.

IgE-independent mast cell activation augments contractility of nonpregnant and pregnant guinea pig myometrium.

BACKGROUND: We have previously shown that in sensitized guinea pigs premature labor can be induced by a type I hypersensitivity reaction. We further hypothesize that premature labor occurs due to increased uterine contractility caused by activation of mast cells and possibly eosinophils, and collective release of their mediators. The objective of this study was to test the hypothesis that IgE-independent mast cell degranulation could increase uterine contractility. METHODS: Longitudinal uterine strips from nonpregnant and pregnant guinea pigs were incubated in organ chambers with vehicle, histamine H(1), serotonin 5-HT(2)/5-HT(1C), thromboxane A(2), leukotriene D(4) receptor antagonists, mast cell stabilizer, and cyclooxygenase or lipoxygenase inhibitors. Then, supernatant, obtained after activation of a mast cell line (MC/9) with compound 48/80, culture medium, or compound 48/80 alone were added. Cumulative concentration-response curves to histamine and serotonin were also obtained. RESULTS: The supernatant and compound 48/80 significantly increased contractility of uterine strips. A mast cell stabilizer considerably reduced the effect of compound 48/80. Other substances attenuated uterine contractile responses to supernatant and compound 48/80, and responses varied depending on the pregnancy period. Histamine and serotonin increased contractility of uterine strips, and uterine sensitivity to these agents were dependent on gestational age. CONCLUSIONS: In summary, mast cells increase uterine contractility through multiple mediators, and uterine responses to these mediators are dependent on gestational age. We postulate that the simultaneous release of these mast cell/eosinophil mediators in the uterus could be a stimulus to trigger and/or maintain myometrial contractions during preterm and term labor.

The effect of a mast cell degranulating agent on vascular resistance in the human placental vascular bed and on the tone of isolated placental vessels.

The objective of this study is to investigate the effect of a mast cell degranulating agent, compound 48/80, on vascular resistance in the perfused human placenta and on the tone of isolated human chorionic vessels. Human placenta was obtained from term nonlaboring women undergoing cesarean delivery. Placental vascular bed perfusion pressure and isometric tension for segments of chorionic plate artery and vein were obtained in response to potassium chloride, compound 48/80, a mast cell stabilizer (cromolyn), and thromboxane A2 mimetic (U46619). Compound 48/80 significantly increased perfusion pressure in isolated human placental cotyledons. This effect was significantly potentiated further after induction of active vascular tone by thromboxane A2 mimetic U46619. Cromolyn significantly attenuated responses to compound 48/80 in these preparations. Compound 48/80 also significantly increased tone in isolated human chorionic artery and vein rings, and responses were abolished by cromolyn. In conclusion, degranulation of placental and intravascular mast cells by compound 48/80 leads to the release of vasoconstrictive substances. This could impair placental blood flow and result in growth restriction in fetuses of women with type l hypersensitivity reactions.

Influence of progesterone on myometrial contractility in pregnant mice treated with lipopolysaccharide.

AIM: To evaluate the effect of progesterone on interleukin (IL)-6, prostaglandin (PG) E2 and nitric oxide (NO) metabolite (NOx) production and contractile activity by NO in pregnant mice treated with lipopolysaccharide (LPS). METHODS: Pregnant C57BL mice on day 14 of gestation were killed 6 h after i.p. injection of LPS (400 microg/kg) or vehicle. Progesterone (2 mg) was subcutaneously injected 2 h before LPS treatment. Uterine rings were equilibrated in Krebs-Henseleit solution (37 degrees C) bubbled with 20% O2 and 5% CO2 (pH 7.4) for sampling and isometric tension recording. IL-6, PGE2 and NOx productions were measured from the bathing solution. Changes in spontaneous contractile activity in response to cumulative concentrations of l-arginine, diethylamine/nitric oxide (DEA/NO, the NO donor), and 8-bromo-cGMP (8-br-cGMP) were compared. Integral contractile activity over 10 min after each concentration was calculated and expressed as percentage change from basal activity. Statistical analyses were performed using one-way anova followed by Dunnett's test (significance was defined as P < 0.05). RESULTS: Interleukin-6 (34.7 +/- 6.0 pg/g tissue), PGE2 (66.8 +/- 6.7 pg/g tissue) and NOx (51.0 +/- 5.4 pmol/2 mL/g wet tissue) production were significantly stimulated by LPS treatment (138.2 +/- 23.2, 147.0 +/- 29.0, 98.6 +/- 16.2, respectively; P < 0.05). L-arginine, DEA/NO and 8-br-cGMP concentration-dependently inhibited spontaneous contractions in uterine rings both in LPS-treated and -untreated animals. Treatment with LPS significantly attenuated the maximal inhibition induced by l-arginine, DEA/NO and 8-br-cGMP in uterine rings from pregnant mice. Progesterone significantly decreased the levels of IL-6 production (74.9 +/- 12.1, P < 0.05), but not PGE2 and NOx production, and contractile responses by l-arginine, DEA/NO and 8-br-cGMP. CONCLUSIONS: The administration of LPS is associated with increases in IL-6, PGE2 and NO, and these increases may or may not have a role to play in LPS-induced preterm labor. Progesterone reduced the LPS-induced increase in IL-6 production and this may be one of the ways that progesterone reduces the risk of preterm labor.

Influence of lipopolysaccharide on the uterine contraction inhibitory effects of tocolytic agents in pregnant mice.

The study was undertaken to investigate the influence of lipopolysaccharide (LPS) on the uterine contraction inhibitory effects of tocolytic agents such as ritodrine, magnesium, and diethylamine/nitric oxide (DEA/NO), and on prostaglandin (PG) E2 and nitric oxide (NO) metabolite (NOx) production in pregnant mice at midgestation. Pregnant C57BL mice on day 14 of gestation were sacrificed 6 hours after intraperitoneal injection of LPS (400 mug/kg) or vehicle. Uterine rings were equilibrated in Krebs-Henseleit solution (37 degrees C) bubbled with 20% O (2) and 5% CO (2) (pH ~7.4) for sampling and isometric tension recording. The concentration levels of PGE2 and NOx in the solution were determined. Changes of spontaneous contractile activity in response to cumulative concentrations of ritodrine, magnesium, and the NO donor, DEA/NO, from the baseline were determined. Integral contractile activity over 10 minutes at each concentration was calculated and expressed as percentage change from basal activity. Statistical analysis was performed using one-way analysis of variance (ANOVA) followed by the Dunnett test (significance: P < 0.05). LPS treatment significantly increased the production levels of PGE2 and NOx (from 66.8 +/- 6.7 pg/g tissue to 147.0 +/- 29.0, from 51.0 +/- 5.4 pmol/2 muL/g tissue to 98.0 +/- 16.2, respectively), P < 0.05). Ritodrine, magnesium, and DEA/NO inhibited spontaneous contractions concentration dependently in uterine rings from both LPS-treated and -untreated animals. Treatment with LPS significantly attenuated the maximal inhibition induced by DEA/NO in uterine rings from pregnant mice. LPS significantly suppressed the uterine contraction inhibitory effects of ritodrine at 10 (-8) M concentrations and of magnesium at 4.2 mmol concentration ( P < 0.05). We concluded that the effects of ritodrine and magnesium may be reduced under inflammatory conditions because LPS increases the production levels of PGE2 and NOx, which cause increased spontaneous contractions of the uterine myometrium. Therefore, when uterine contractions are not controlled by tocolytics in pregnant patients with preterm labor associated with inflammation, labor induction or pregnancy termination may become significant options in clinical practice.

PAR-2 activating peptide-induced stimulation of pregnant rat myometrium contractile activity partly involves the other membrane receptors.

OBJECTIVE: To study if spontaneous contractions augmented by proteinase-activated receptor-2 (PAR-2)-activating peptide serine-leucine-isoleucine-glycine-arginine-leucine (SLIGRL) involve coactivation of membrane chemoceptors and are associated with expression of PAR-2 mRNA in non-pregnant and pregnant rat myometrium. MATERIALS AND METHODS: Non-pregnant, mid-pregnant, and late pregnant rat uterine horn and small intestine segments were snap-frozen in liquid nitrogen to determine PAR-2 mRNA levels by real time polymerase chain reaction (PCR). Uterine rings were used for isometric tension recording. Effect of SLIGRL (0.1 mM) on spontaneous contractions before and after exposure to ibuprofen (cyclooxygenase inhibitor, 1.0 microM), SQ-29548 (thromboxane A(2) receptor inhibitor, 1.0 microM), ketotifen (histamine 1 receptor inhibitor, 10 microM), WEB-2170BS (platelet-activating factor (PAF) receptor inhibitor, 10 microM), atropine (muscarinic receptor inhibitor, 0.1 microM), or ketanserin (serotonin receptor inhibitor, 10 microM) were compared. Paired t-test and one-way ANOVA followed by Dunnett's or Newman-Keuls post hoc tests were used for statistical analysis when appropriate. SIGNIFICANCE: P<0.05. RESULTS: The agents did not significantly affect time-associated decay in spontaneous contractile activity in any group of the tissues. Activation of spontaneous contractions induced by SLIGRL in non-pregnant rat myometrium did not involve coactivation of membrane chemoceptors, while in mid-pregnant rat myometrium coactivation of prostanoid, histamine, and serotonin receptors and in late pregnant rat myometrium coactivation of thromboxane receptors was noted. Expression of PAR-2 mRNA was similar in non-pregnant, mid-pregnant, and late pregnant rat myometrium. CONCLUSIONS: Expression of PAR-2 in rat myometrium is not dependent on gestational age. Stimulation of PAR-2 is associated with production/release of cyclooxygenase pathway product(s) activating thromboxane/prostaglandin H2 receptors, partial involvement of histamine H1 receptors and serotonin receptors in midpregnancy and thromboxane A2/prostaglandin H2 receptors in late pregnancy.

Adaptation to pregnancy leads to attenuated rat uterine artery smooth muscle sensitivity to oxytocin.

OBJECTIVE: We tested the hypothesis that oxytocin (OT) contracts blood vessels via vasopressin V1A (VP) receptors, and that this depends on pregnancy. STUDY DESIGN: Concentration-contraction relationships (CCR) to OT and VP (10(-12)-10(-6) mol/L) were obtained in different blood vessels. CCR were obtained in uterine arteries (UA) from nonpregnant, mid-pregnant, and late pregnant rats (n = 6-10 per group) in the absence and presence of selective antagonists (10(-7) mol/L). RESULTS: Sensitivity to OT, but not to VP, is attenuated in pregnant rat UA. Antagonists shifted CCR of OT and VP to the right, and, to a lesser extent, of the counterpart, in all UA. VP antagonist depresses oxytocin CCR much more than OT antagonist in pregnant rat UA. CONCLUSION: OT and VP contract UA via their own receptors, although partial cross-activation is evident. Adaptation to pregnancy led to attenuated sensitivity of UA smooth muscle to OT and transformed OT receptors into VP-like ones.

Fetal origins of adult vascular dysfunction in mice lacking endothelial nitric oxide synthase.

Epidemiological studies have shown increased incidence of hypertension and coronary artery disease in growth-restricted fetuses during their adult life. A novel animal model was used to test the hypothesis regarding the role of an abnormal uterine environment in fetal programming of adult vascular dysfunction. Mice lacking a functional endothelial nitric oxide synthase (NOS3-/-KO, where KO is knockout) and wild-type (WT) mice (NOS3+/+WT) were crossbred to produce homozygous NOS3-/-KO, maternally derived heterozygous (NOS3+/-mat, mother with NOS3 deficiency), paternally derived heterozygous (NOS3+/-pat, normal mother), and NOS3+/+WT litters. Number of fetuses per litter was smaller in NOS3-/-KO and NOS3+/-mat compared with NOS3+/-pat and NOS3+/+WT mice. Adult female mice from these litters (7-8 wk old) were killed, and ring preparations of carotid and mesenteric arteries were mounted in a wire myograph to evaluate the passive and reactive vascular characteristics. Slope of the length-tension plot (a measure of vascular compliance) was increased, and optimal diameter (as calculated by Laplace equation) was decreased in NOS3-/-KO and NOS3+/-mat compared with NOS3+/-pat and NOS3+/+WT mice. Acetylcholine caused vasorelaxation in NOS3+/-pat and NOS3+/+WT and contraction in NOS3-/-KO and NOS3+/-mat mice. Responses to phenylephrine and Ca2+ were increased in NOS3-/-KO and NOS3+/-mat compared with NOS3+/-pat and NOS3+/+WT mice. Relaxation to isoproterenol was decreased in NOS3-/-KO and NOS3+/-mat vs. NOS3+/-pat and NOS3+/+WT mice. Abnormalities in the passive and reactive in vitro vascular properties seen in NOS+/-mat that developed in a NOS3-deficient maternal/uterine environment compared with the genetically identical NOS3+/-pat mice that developed in a normal environment are the first direct evidence in support of a role for uterine environment in determining vascular function in later life.

Degranulation of uterine mast cell modifies contractility of isolated myometrium from pregnant women.

OBJECTIVE: This study was undertaken to test that uterine mast cell degranulation alters human myometrial contractility in vitro and to define what mediators are involved in this process. STUDY DESIGN: Longitudinal myometrial strips prepared from biopsy specimen obtained from the lower uterine segment of women at preterm and term gestation (with and without labor) were studied. Contractile responses to compound 48/80, a mast cell degranulator, were compared in the absence or presence of a mast cell stabilizer, H 1 and H 2 receptor antagonists, cyclooxygenase, and lipoxygenase inhibitors. RESULTS: Compound 48/80 increased myometrial contractility in all groups. The mast cell stabilizer cromolyn inhibited contractility, whereas the cyclooxygenase inhibitor ibuprofen, the H 1 -receptor antagonist S(+)-chlorpheniramine maleate, but not the H 2 antagonist cimetidine, only slightly attenuated this effect. The lipoxygenase inhibitor linoleyl hydroxamic acid augmented the responses to compound 48/80 in the preterm but not in the term group. CONCLUSION: Uterine mast cell degranulation, or the effects of their mediators, can modulate uterine contractility during pregnancy.

Induction of premature labor and delivery by allergic reaction and prevention by histamine H1 receptor antagonist.

OBJECTIVE: The purpose of this study was to determine whether type I hypersensitivity can induce preterm labor and delivery that may be prevented by the administration of an H 1 receptor antagonist or a mast-cell stabilizer. STUDY DESIGN: At days 40 through 50 of gestation, ovalbumin-sensitized guinea pigs that had not been treated or had been pretreated with an H 1 receptor antagonist (ketotifen) or a mast-cell stabilizer (cromolyn sodium) were challenged with either ovalbumin or saline solution. Nonsensitized guinea pigs were challenged with ovalbumin. RESULTS: The duration of gestation was significantly shorter in sensitized animals that were challenged with ovalbumin than in the other groups. Pretreatment with ketotifen significantly increased the duration of pregnancy in sensitized animals, compared with untreated animals. Cromolyn sodium had no effect. CONCLUSION: An allergic reaction can induce premature labor/delivery, and pretreatment with histamine H 1 receptor antagonist can prevent it. These observations provide evidence of a novel mechanism of disease for premature labor and delivery.

Enhanced growth and improved vascular function in offspring from successive pregnancies in endothelial nitric oxide synthase knockout mice.

OBJECTIVE: Transgenic mice that lack endothelial nitric oxide synthase have offspring with growth deficiency and abnormal vascular reactivity in later life. Our objective was to evaluate the role of parity in the modulation of the fetal programming of growth and vascular responses in these transgenic mice. STUDY DESIGN: Oligoparous (0-2 previous pregnancies) and multiparous (5-9 previous pregnancies) nitric oxide synthase knockout (-/-KO) female mice were bred with nitric oxide synthase -/-KO and wild type (+/+WT) male mice to produce nitric oxide synthase -/-KO and maternally derived heterozygous (+/-Mat) litters. The pups were weighed weekly. Carotid arteries of the adult females from these litters were used for in vitro vascular reactivity studies. RESULTS: Nitric oxide synthase knockout and nitric oxide synthase maternal litters that were born to oligoparous mothers had significant growth lag compared with corresponding multiparous litters. Length-tension characteristics were not different between the groups. However, optimal diameter, which is a measure of vascular tensile properties and resistance, was decreased in oligoparous compared with multiparous female offspring. Acetylcholine-mediated vasorelaxation was abolished, and contraction by phenylephrine and Ca ++ was increased in oligoparous, but not multiparous, female offspring ( P < .05). CONCLUSION: These data support the role of abnormal uterine environment in the fetal programming of postnatal growth and vascular function in later life. Successive pregnancies may lead to maternal uterine adaptations that bypass the lack of a functional nitric oxide synthase, which leads to improvement in postnatal growth and vascular function in the offspring. Given the reported effect of parity on the risk of preeclampsia, similar mechanisms may be operative in human pregnancy.

Effects of nitric oxide donors on non-pregnant and pregnant rat uterine and aortic smooth muscle.

OBJECTIVE: To compare the effects of nitric oxide (NO) donors, diethylamine/nitric oxide (DEA/NO) and nitroglycerin (NTG), on isolated uterine and aortic tissues from non-pregnant, mid and late pregnant rats. METHODS: The uterus and thoracic aorta were obtained from non-pregnant (estrous cycle) and pregnant Sprague-Dawley rats on day 14 and day 21. The uterine and aortic rings were incubated in organ chambers filled with Krebs-Henseleit solution bubbled with 5% CO2 in air for isometric tension recordings. Cumulative concentration-response relationships to DEA/NO and NTG were obtained in the aortic rings contracted with phenylephrine and in spontaneously contracting uterine rings. RESULTS: The sensitivity and the maximal inhibitory effects of DEA/NO did not differ in aortic tissues of any group. DEA/NO-induced Maximal inhibition of spontaneous contractions of uterine tissues from mid-pregnant rats was greater (although not significantly) than in the tissues from non-pregnant animals (with similar sensitivity), but it was significantly depressed in tissues from late pregnant rats. The sensitivity to and maximal inhibitory effects of NTG were less in aortic tissues from late pregnant versus mid-pregnant and non-pregnant rats. In uterine tissues from late pregnant rats the effect of NTG was negligible. The inhibitory action of both NO donors was much more pronounced in aortic versus uterine tissues. CONCLUSIONS: Uterine smooth muscle is less sensitive than vascular smooth muscle to NO. Uterine smooth muscle from late pregnant animals demonstrates refractoriness to both DEA/NO and NTG, while vascular smooth muscle from late pregnant animals demonstrates refractoriness to NTG, but not to DEA/NO.

Effects of recombinant human relaxin on pregnant rat uterine artery and myometrium in vitro.

OBJECTIVE: The purpose of this study was to evaluate the effects of recombinant human relaxin on the uterine artery and myometrial contractility in pregnant rats. STUDY DESIGN: Uterine artery and myometrial rings from mid and term pregnant rats were used. Relaxin effect was studied on phenylephrine-induced contraction in the presence or absence of nitric oxide synthase inhibitor, N omega-nitro-l-arginine methyl ester, soluble guanylate cyclase inhibitor, 1H-oxadiazolo-quinoxaline-1-one, or adenylate cyclase inhibitor, SQ-22,536. The myometrial inhibitory effect of relaxin was studied on spontaneous and oxytocin- or protein kinase C activator-induced contractions. RESULTS: Uterine artery relaxation by relaxin was greater at mid pregnancy compared with term. Relaxin effect was decreased by SQ-22,536, 1H-oxadiazolo-quinoxaline-1-one and N omega-nitro-l-arginine methyl ester at mid pregnancy. Relaxin inhibited spontaneous contractions at mid pregnancy but not at term. Relaxin had no effect on oxytocin- or indolactam-V-induced contractions. CONCLUSION: Relaxin effect is mediated by nitric oxide, soluble guanylate cyclase, and adenylate cyclase in mid pregnant uterine artery. Relaxin inhibits spontaneous uterine activity at mid pregnancy. Relaxin effect decreased at term gestation in both tissues.

Proteinase-activated receptor-2 activation induces uterine contractility in term pregnant rats that is not dependent on mast cell activation and cyclo-oxygenase products.

OBJECTIVES: This study was undertaken to evaluate the effect of proteinase-activated receptor-2 (PAR-2) activation on the contractility of uterine tissues from term pregnant rats and the role of mast cells and prostaglandins in such an effect. STUDY DESIGN: Uterine rings from pregnant (day 20-21) Sprague-Dawley rats were used for isometric tension recording in organ chamber experiments (Krebs solution, 5% carbon dioxide in air, 37 degrees C, pH approximately 7.4). Responses to the PAR-2 activating peptide SLIGRL (serine-leucine-isoleucine-glycine-arginine-leucine), and to the inactive reverse peptide LRGILS (leucine-arginine-glycine-isoleucine-leucine-serine) were determined after pretreatments with compound 48/80, cromolyn, S[+]-chlorpheniramine maleate, cimetidine, combinations of histamine (H) receptor antagonists with cromolyn or ibuprofen and compared with vehicle. RESULTS: SLIGRL significantly augmented contractility of uterine tissues, and this response was not inhibited by compound 48/80, cromolyn, and ibuprofen, as well as by H(1)- and H(2)-receptor antagonists, alone or in combination with cromolyn. CONCLUSION: PAR-2 activation augments uterine contractility in tissues obtained from term pregnant rats, and this effect is independent of mast cell activation or cyclo-oxygenase pathway products.

Sex hormone effects in non-pregnant rat and human myometrium.

OBJECTIVE: To study effects of sex hormones on spontaneous contractility and on the effects of depolarizing agent potassium chloride (KCl), M-cholinoceptor and prostaglandin receptor agonists on non-pregnant rat and human uterine tissues. STUDY DESIGN: Uterine rings from ovariectomized rats treated with sex hormones or placebo, and uterine strips from premenopausal and postmenopausal women were equilibrated in Krebs buffer (t=37 degrees C, pH approximately 7.4) for isometric tension recordings. Spontaneous contractile activity and contractions in response to KCl, eicosanoids, and acetylcholine were compared. RESULTS: In tissues from ovariectomized rats, spontaneous contractility was increased, while KCl-induced contractions were decreased. Treatment with 17beta-estradiol, but not progesterone, inhibited spontaneous contractions, but potentiated KCl evoked contractions. Treatment with 17beta-estradiol did not influence responses to prostanoids in ovariectomized rats, while treatment with both sex hormones restored decreased the responses. Spontaneous contractility and responses to KCl were less in uterine tissues from postmenopausal versus premenopausal women. Hormone replacement therapy partly restored the responses to KCl, prostanoids, and acetylcholine. CONCLUSIONS: Ovarian steroids modulate spontaneous contractile activity, responses to depolarization, prostanoids and M-cholinoceptor activation in non-pregnant rat and human uterine tissues in vitro.

Effect of histamine on phasic and tonic contractions of isolated uterine tissue from pregnant women.

OBJECTIVE: To enhance our understanding of the uterotonic effect of histamine, we compared the effects of histamine on spontaneous phasic and tonic contractile activity of uterine strips from term pregnant nonlaboring women. STUDY DESIGN: Longitudinal uterine strips were used from the lower uterine segment of term pregnant nonlaboring women undergoing elective cesarean section. The concentration-response relationship to histamine (10(-8) to 10(-4) mol/L) was determined in spontaneously contracting strips or in strips contracted tonically with a protein kinase C activator (-)-indolactam V in the presence of H(1) receptor antagonist (S[+]-chlorpheniramine maleate), H(2) receptor antagonist (cimetidine), or solvent. RESULTS: Histamine increased spontaneous phasic myometrial contractions in a concentration-dependent manner. H(1), but not H(2), receptor antagonist significantly attenuated the response to histamine. Histamine significantly reduced tonic contractions of uterine strips induced by indolactam V. H(1) histamine receptor antagonist abolished relaxation, whereas H(2) histamine receptor antagonist had no effect. CONCLUSION: Histamine increases spontaneous, but inhibits tonic, contractions of uterine strips from term pregnant nonlaboring women. Both effects are mediated through activation of H(1) receptors.

Flow rate-perfusion pressure relationships in situ in the uterine circulation of pregnant rats.

OBJECTIVE: Our purpose was to study the effects of inhibition of nitric oxide synthesis on perfusion pressure and flow rate-perfusion pressure relationships in the rat uterine circulation in situ. STUDY DESIGN: Nonpregnant, midpregnant (day 14), and late pregnant (day 21) Sprague-Dawley rats were studied. The vascular bed of the intact uterus and its contents were isolated and perfused with Krebs buffer (37 degrees C, pH approximately 7.4, 2% dextran and indomethacin, 10(-5) mol/L) through a cannula inserted into the abdominal aorta close to the iliac artery bifurcation, and perfusion pressure was monitored. After equilibration, the flow rate was increased from 1 mL/min to 8 or 16 mL/min, in the absence and presence of N(omega)-nitro-L-arginine methyl ester (L-NAME), phenylephrine, or both. RESULTS: The flow rate-perfusion pressure relationship in midpregnant rats (n = 9) was not significantly different from that in late pregnant rats (n = 12), but the latter was significantly greater than the relationship in nonpregnant animals (n = 5). L-NAME did not influence perfusion pressure and flow rate-perfusion pressure relationships in any of the groups. However, L-NAME enhanced the phenylephrine-induced and flow rate-induced increase in perfusion pressure in the vascular beds from nonpregnant and midpregnant animals, and to a lesser extent in late pregnant rats. L-arginine did not influence perfusion pressure or the flow rate-perfusion pressure relationship in any group. CONCLUSIONS: Vasoconstriction produced by phenylephrine uncovers basal release of endothelium derived nitric oxide. Vasoconstriction increases perfusion pressure responses to increases in flow rate in the uterine vascular beds of nonpregnant, midpregnant, and late pregnant rats. The release of endothelial nitric oxide in the uterine vascular beds depends on the basal contractile state of the vasculature.

Endogenous mast cell degranulation modulates cervical contractility in the guinea pig.

OBJECTIVE: The purpose of this study was to investigate the effect of endogenous mast cell degranulation on the contractility of isolated cervical strips from nonpregnant and pregnant guinea pigs. STUDY DESIGN: Longitudinal cervical strips from nonpregnant and pregnant (mid and term) guinea pigs were used for isometric tension recording. Responses to the mast cell degranulating agent, compound 48/80, were compared in the absence or presence of different inhibitors and receptor antagonists. Concentration-response curves were obtained to histamine and 5-hydroxytryptamine in strips that were incubated with antagonists or solvent. RESULTS: Compound 48/80 and histamine significantly increased contractility of cervical strips in all 3 groups of animals. The inhibitor of mast cell degranulation significantly reduced responses to compound 48/80 and histamine-1 receptor antagonist reduced responses to histamine in all 3 groups. Histamine-1 receptor antagonist significantly inhibited responses to compound 48/80 in nonpregnant and mid pregnant guinea pigs. Histamine-2 receptor antagonist did not alter responses to compound 48/80 nor to histamine. The receptor antagonist 5-hydroxytryptamine-2 significantly inhibited cervical contractility that was induced by compound 48/80 in tissues from mid pregnant and term pregnant guinea pigs. Lipoxygenase inhibitor was effective in mid pregnant guinea pigs. Cyclooxygenase inhibitor, 5-hydroxytryptamine, and a combination of lipoxygenase and cyclooxygenase inhibitors had no effect on cervical contractility. CONCLUSION: The degranulation of mast cells releases histamine and other mediators that stimulate cervical contractility through histamine-1 receptors. Cervical infiltration and modulation of contractility by mast cells may play an important physiologic and/or pathologic role in the control of cervical function during pregnancy.

Prostaglandin-induced activation of uterine contractility in pregnant rats does not involve potassium channels.

OBJECTIVE: The uterus is a target for prostaglandins, especially at the end of gestation. Whether potassium channels are involved in the effect of prostaglandins is not clear. The aim of this study was to find out. STUDY DESIGN: Concentration-response relationships to prostaglandins (prostaglandin F2alpha, prostaglandin E2, and prostaglandin I2 [carbacyclin]; 10(-10) mol/L-10(-4) mol/L) were studied in isolated uterine rings from mid pregnancy (day 14) and late pregnancy (day 21) rats (Krebs solution, 5% CO2 in air, 37 degrees C; pH, 7.4). Rings were incubated for 30 minutes with either solvent or adenosine triphosphate-sensitive potassium channel inhibitor or opener glibenclamide and levcromakalim or with calcium-sensitive potassium channel inhibitor or opener NS1619 and iberiotoxin, respectively. The changes in integral activity were compared after each concentration of the agent and were expressed as a percent of the basal integral activity. RESULTS: The increases in spontaneous contractile activity induced by prostaglandin E2 and carbacyclin, but not prostaglandin F2alpha, were statistically significantly higher in tissues from late pregnancy versus mid pregnancy rats and were not affected by any of the K-channel openers or inhibitors. CONCLUSION: Adenosine triphosphate-sensitive and calcium-sensitive potassium channels are not involved in the effect of prostaglandin F2alpha, prostaglandin E2, and prostaglandin I2 on pregnant rat uterus.