Vitamin C supplementation modulates crude oil contaminated water induced gravid uterine impaired contractile mechanism and foetal outcomes in Wistar rats.

OBJECTIVES: Crude oil is a common environmental contaminant that impacts the reproductive functions of women. Understanding the contractile mechanism of the gravid uterus and how it impacts fetal outcomes during crude oil-contaminated water (CCW) exposure is still evolving. This study investigates the effect of vitamin C supplementation during the ingestion of CCW from Bayelsa, Nigeria, on the contractile mechanism of the gravid uterus and fetal outcomes. METHODS: Fifteen nulliparous pregnant rats were randomly divided into 3 groups of 5 rats each and treated with normal saline (control), CCW (2.5 mL), and CCW + vitamin C (10 mg/kg bwt), respectively. Treatments were via oral gavage from gestation days 1-19. Gas chromatography-mass spectrometry of CCW, uterine oxidative biomarkers, and in vitro contractile activity of excised uterine tissue to acetylcholine, oxytocin, magnesium, and potassium were determined. Furthermore, uterine responses to acetylcholine after incubation with nifedipine, indomethacin, and N-nitro-L-arginine methyl ester were also recorded using the Ugo Basile data capsule acquisition system. Fetal weights, morphometric indices, and anogenital distance were also determined. RESULTS: Acetylcholine, oxytocin, magnesium, diclofenac, and indomethacin-mediated contractile mechanisms were significantly impaired with CCW exposure; however, vitamin C supplementation significantly attenuated the impaired uterine contractile activity. Maternal serum estrogen, weight, uterine superoxide dismutase, fetal weight, and anogenital distance were significantly reduced in the CCW group compared to the vitamin C supplemented group. CONCLUSIONS: Ingestion of CCW impaired the uterine contractile mechanism, fetal developmental indices, oxidative biomarkers, and estrogen. Vitamin C supplementation modulated these by elevating uterine antioxidant enzymes and reducing free radicals.

Obesity alters the uterine environment before pregnancy.

Obesity is a metabolic disorder that predisposes to numerous diseases and has become a major global public health concern. Cafeteria diet (CAF) is the animal model used for the study of obesity that more closely reflects Western diet habits. Previously, we described that CAF administration for 60 days induces obesity in female rats and their fetuses develop macrosomia. Given that, in our model, rats are not genetically modified and that obese mothers were fed standard chow during pregnancy, the aim of the current study was to test the hypothesis that obesity alters the intrauterine environment prior to pregnancy, and this may explain the exacerbated fetal weight gain. We found that uteri from obese rats during the estrous phase developed insulin resistance through mechanisms that involve the induction of uterine hypoxia and the down-regulation of the insulin receptor gene. Moreover, uterine cell proliferation was induced by obesity concomitantly with the reduction in the uterine contractile response to a ß2 AR agonist, salbutamol, and this may be consequence of the down-regulation in the uterine ß2 AR expression. We conclude that CAF-induced obesity alters the uterine environment in rats during the estrous phase and may cause the fetal macrosomia previously described by us in obese animals. The lower sensitivity of the uterus to a relaxation stimulus (salbutamol) is not a minor fact given that for implantation to occur the uterus must be relaxed for embryo nidation. Thus, the alteration in the uterine quiescence may impair implantation and, consequently, the foregoing pregnancy.

Leukotriene receptor antagonist as a novel tocolytic in an in vitro model of human uterine contractility.

OBJECTIVE: This study analyzed the ability of montelukast, a cysteinyl-leukotrienes receptor antagonist and anti-inflammatory agent, to produce a consistent tocolytic effect alone or in combination with nifedipine, a calcium (Ca(2+)) channel blocker currently used in clinical practice. STUDY DESIGN: Uterine biopsies were obtained from consenting women undergoing elective cesarean sections at term (n=20). Myometrial microsomal fractions were analyzed by immunoblotting to quantify relative cysteinyl leukotrienes receptor 1 (CysLTR1) levels. Isometric tension measurements were performed in vitro on human myometrial strips (n=120) in isolated organ baths in order to establish concentration-response curves to montelukast and to quantify changes in Ca(2+) sensitivity on ß-escin permeabilized tissues. RESULTS: Immunodetection analysis revealed the presence of CysLTR1 receptor in uterine tissues, fetal membranes and placenta. A significant increase in area under the curve (AUC) was quantified following the addition of leukotriene D4 (LTD4) (0.01-0.3 µM), an end-product of the lipoxygenase pathway. Conversely, addition of montelukast produced a significant tocolytic effect by decreasing the frequency and AUC (IC50=1 µM). Moreover, addition of montelukast also resulted in a reduced Ca(2+) sensitivity as compared to control tissues (EC50 values of 654 and 403 nM; p=0.02 at pCa 6), while an additive effect was observed in combination with 0.1 nM nifedipine (p=0.004). CONCLUSION: This original study demonstrates the potency of montelukast as a tocolytic agent in an in vitro human uterine model. Montelukast, in combination with nifedipine, could represent a therapeutic approach to reduce inflammation associated with prematurity while facilitating the inhibition of preterm labor.

Effect of cytochrome P-450 epoxygenase and hydroxylase metabolites on rat myometrium contractility in non-pregnancy, late pregnancy and late pregnancy under inflammatory conditions.

AIM: The aim of the present experimental study was to assess the tocolytic effect of eicosanoids on myometrium from non-pregnant and pregnant rats with or without an induced inflammatory condition. METHODS: Three hundred myometrial rings were obtained by median laparotomy from 50 Sprague-Dawley rats divided into three groups: (i) non-pregnant (n = 15); (ii) pregnant in absence (n = 20); or (iii) pregnant in presence (n = 15) of lipopolysaccharide treatment, timed at 22 days of pregnancy. Spontaneous contractile activities were compared by isometric tension measurements. The effects of epoxy- and hydroxyeicosanoids derived from arachidonic acid as well as specific enzyme inhibitors were assessed. Changes were expressed as percentage of basal activity by calculating the area under the curve as a function of drug concentration and compared to the effect of the vehicle. RESULTS: A decrease in contractile activity ranging 10-25% was observed upon addition of epoxy- and hydroxyeicosanoids. Increasing epoxyeicosanoid bioavailability by inhibiting their degradation induced a tocolytic effect in the non-pregnant group (20%) and in inflammation-induced condition (40%). There was a significant difference in reactivity between groups and pregnancy condition. Semiquantification of metabolic enzymes that produce (cytochrome P-450 epoxygenase) and degrade (soluble epoxide hydrolase) epoxyeicosanoids by western blot analysis revealed that these enzymes were mainly detected in the non-pregnant group. CONCLUSION: Eicosanoids can modify myometrial reactivity and their presence and effects are amplified in non-pregnant and in inflammation-induced condition. Our data suggest that in contrast to prostaglandins, epoxyeicosatrienoic acids are likely involved in the quiescence phase of parturition because they reduce the rhythmic contractile activity of uterine tissues in pregnant rats.

The effects of etomidate on the contraction of pregnant rat uterine smooth muscle.

BACKGROUND: It has been reported that etomidate has the relaxant effects on vascular, tracheal, and non-pregnant uterine smooth muscle in vitro. The purpose of this study was to investigate the relaxant effects of etomidate on the contraction of the pregnant rat uterine smooth muscle. METHODS: Uterine muscle tissues were obtained from pregnant rats (n = 15). The uterine segments were mounted in organ baths filled with Krebs solution. After oxytocin-induced contractile activity had been established, etomidate in incremental concentrations (10(-7) to 10(-3) M) was added cumulatively to the bath, each administered 20 min apart, and resultant changes in contractile activity were continuously recorded. EC5 (effective concentration of 5% reduction), EC25, EC50, EC75, and EC95 on active tension were calculated using a probit model. RESULTS: Etomidate (10(-7) to 10(-3) M) induced dose-dependent decreases in amplitude and frequency of uterine contraction. The EC50 of etomidate on active tension were 5.91 x 10(-5) M. CONCLUSIONS: These results demonstrate that etomidate had inhibitory effects on pregnant rat uterine muscle at supraclinical concentration (5.91 x 10(-5) M).