Myometrial Contractions Research Articles

Activation of prostaglandin EP4 receptor attenuates the induction of cyclooxygenase-2 expression by EP2 receptor activation in human amnion fibroblasts: implications for parturition.

Human amnion fibroblasts produce abundant prostaglandin E2 (PGE2), which plays a crucial role in parturition by stimulating not only myometrial contraction and cervical ripening but also the expression of the rate-limiting enzyme in PGE2 synthesis-namely, cyclooxygenase-2 (COX-2). This feed-forward induction of COX-2 expression by PGE2 is mediated via its receptors coupled with the cAMP and PKA pathway and subsequent phosphorylation of the transcription factors cAMP-response element binding protein (CREB) and signal transducer and activator of transcription 3 (STAT3). Although prostaglandin E receptor (EP)-2 and EP4 for PGE2 are coupled with activation of the cAMP and PKA pathway, the exact roles of these 2 receptors in the regulation of COX-2 expression in amnion fibroblasts remain to be determined. Here, we clarify this issue by employing human amnion tissue and fibroblasts with the long-term objective of specific targeting of prostaglandin synthesis in prevention of preterm birth. We find that an EP2 agonist caused long-lasting increases in CREB phosphorylation and COX-2 expression, whereas an EP4 agonist induced only transient increases in CREB phosphorylation and COX-2 expression in amnion fibroblasts. Moreover, only EP2 stimulation increased STAT3 phosphorylation, whereas only EP4 stimulation increased PI3K activity. EP4 antagonist or inhibition of PI3K enhanced the induction of CREB and STAT3 phosphorylation and COX-2 expression by PGE2 or EP2 stimulation, which was attenuated by EP4 overexpression. Of interest, PGE2 and cortisol, both well-demonstrated stimulants of COX-2 expression in amnion fibroblasts, increased EP2 but decreased EP4 receptor expression. Furthermore, increased EP2 but decreased EP4 abundance were observed in amnion tissue at parturition. We conclude that EP2 and EP4 receptors play different roles in the regulation of COX-2 expression in human amnion fibroblasts. EP2 is the dominant PGE2 receptor mediating the induction of COX-2 at parturition, which can be attenuated by simultaneous activation of PI3K coupled to the EP4 receptor.-Lu, J.-W., Wang, W.-S., Zhou, Q., Gan, X.-W., Myatt, L., Sun, K. Activation of prostaglandin EP4 receptor attenuates the induction of cyclooxygenase-2 expression by EP2 receptor activation in human amnion fibroblasts: implications for parturition.

GIT2 deficiency attenuates inflammation-induced expression of pro-labor mediators in human amnion and myometrial cells†.

Untimely activation of the inflammatory response by sterile or infective insults in uterine tissues can result in preterm birth. Pro-inflammatory cytokines and pathogenic activation of toll-like receptors (TLRs) initiate a biochemical cascade of events leading to myometrial activation and contractility, cervical dilatation, and rupture of the chorioamniotic membranes. GIT2 is a signaling protein known to play a role in innate and adaptive immunity; however, its role in the inflammatory pathways of human labor is not known. In this article, we report that GIT2 expression is lower in human myometrium and fetal membranes with term labor, and in preterm amnion with histological chorioamnionitis. GIT2 knockdown by siRNA in primary myometrial and amnion cells exhibited reduced expression of pro-inflammatory cytokines and chemokines in response to inflammatory challenge by cytokines or TLR ligands. In addition, the pro-inflammatory cytokines IL1B and TNF could not induce the expression of extracellular matrix degrading enzymes in GIT2-deficient amnion cells. Myometrial activation in response to pro-inflammatory cytokines was also significantly suppressed in GIT2-deficient cells as evidenced by decreased prostaglandin release and expression of contraction-associated proteins. Further to this, collagen gel assays demonstrated that TNF had a reduced ability to induce myometrial contractility in situ in GIT2-deficient myometrial cells compared to control-transfected cells. In summary, the loss of GIT2 diminishes the effects inflammatory mediators have in promoting myometrial contraction and fetal membrane rupture in vitro, suggesting that GIT2 could be a possible target for preterm birth therapies.

Role of biologically active molecules in uterine contractile activity

Hypothesis/aims of study. Myometrial relaxation and contraction require synchronous cellular interactions. At present, it has been established that the coordination of myometrial contractile activity is carried out by a conduction system constructed from gap junctions with intercellular channels. There are no clinical data on inhibiting (nitric oxide synthase) and activating (connexin-43) factors of uterine contractile activity in the myometrium during pregnancy and parturition in the published literature. This study was undertaken to measure the expression levels of nitric oxide synthase, adhesion molecules CD51, CD61, and connexin-43 in the myometrium during pregnancy and parturition; and to assess the role of inhibitory and activating factors in the development of uterine contractile activity.
 Study design, materials and methods. An immunohistochemical study of myometrial biopsy specimens obtained from the lower uterus segment during cesarean section was performed in eight women with a full-term physiological pregnancy, in another eight individuals in the active phase of uncomplicated parturition, and in eight patients with uterine inertia. Integrins (CD51 and CD61 proteins) were used as markers of cell adhesion. Localization and the number of intercellular contacts were assessed by measuring the expression level of connexin-43, with the intensity of oxidative processes assessed by nitric oxide synthase activity.
 Results. In the myometrium, in the active phase of physiological parturition, a three-fold increase in the expression of activating (CD51, CD61, and connexin-43) factors of uterine contractile activity and a five-fold decrease in that of inhibitory (nitric oxide synthase) ones occur compared to those in full-term physiological pregnancy.
 Conclusion. In the pathogenesis of uterine inertia and resistance to labor induction, an important role is played by the decreased expression of adhesion molecules (CD51, CD61) and connexin-43 and the increased expression of nitric oxide synthase in the myometrium.

Effect of Inclined Magnetic Field on Peristaltic Flow of Carreau Fluid through Porous Medium in an Inclined Tapered Asymmetric Channel

During this article, we have a tendency to show the peristaltic activity of magnetohydrodynamics flow of carreau fluid with heat transfer influence in an inclined tapered asymmetric channel through porous medium by exploitation the influence of non-slip boundary conditions. The tapered asymmetric channel is often created because of the intrauterine fluid flow induced by myometrial contraction and it had been simulated by asymmetric peristaltic fluid flow in an exceedingly two dimensional infinite non uniform channel, this fluid is known as hereby carreau fluid, conjointly we are able to say that one amongst carreau's applications is that the blood flow within the body of human. Industrial field, silicon oil is an example of carreau fluid. By exploitation, the perturbation technique for little values of weissenberg number, the nonlinear governing equations in the two-dimensional Cartesian coordinate system is resolved under the assumptions of long wavelength and low Reynolds number. The expressions of stream function, temperature distribution, the coefficient of heat transfer, frictional forces at the walls of the channel, pressure gradient are calculated. The effectiveness of interesting parameters on the inflow has been colluded and studied.

Toll-like receptor 9, maternal cell-free DNA and myometrial cell response to CpG oligodeoxynucleotide stimulation.

Among mechanisms triggering onset of parturition, it has been recently postulated that Toll-Like Receptor (TLR)9 engagement by cell-free DNA (cfDNA) triggers inflammation, myometrial contractions, and labor in absence of infection. The current study evaluated whether direct (myometrial) or indirect (decidual) TLR9 engagement enhances human myometrial contractility. Toll-like receptor 9 expression and cellular localization were surveyed by immunohistochemistry of placenta, fetal membranes, and myometrium in term (gestational age [GA]: >37weeks) labor (TL, n=7) or term non-labor (TNL, n=7) tissues. Non-pregnant myometrium (n=4) served as reference. TLR9 mRNA expression relative to other TLRs was evaluated through the mining of an RNA-seq dataset and confirmed by RT-PCR. Immortalized human myometrial cells (hTERT-HM) were treated with incremental concentrations of TLR9 agonist ODN2395, TNF-α, or LPS. Secreted cytokines were quantified by multiplex immunoassay, and contractility was assessed by an in-gel cell contraction assay (n=9). Induction of hTERT-HM contractility was also evaluated indirectly following exposure to conditioned media from primary term decidual cells (n=4) previously stimulated with ODN2395. Toll-like receptor 9 immunostaining in placenta and amniochorion was strongest in decidual cells, but unrelated to labor. TLR9 staining intensity was significantly decreased in TL compared with TNL myometrium (P=0.002). Although total cfDNA in maternal circulation increased in TL (P=0.025 vs TNL), difference in cffDNA was non-significant. Myometrial TLR9 mRNA levels were unaffected by contractile status and far less abundant than other pro-inflammatory TLRs. hTERT-HM contractility was enhanced by LPS (P=0.002) and TNF-α (P=0.003), but not by ODN2395 (P=0.345) or supernatant of TLR9-stimulated decidual cells. Myometrial and decidual TLR9 are unlikely to directly regulate human parturition.

Computer modelling of peristalsis-driven intrauterine fluid flow in the presence of electromagnetohydrodynamics

The intrauterine fluid motion is responsible for embryo transport and its implantation at fundus. It is believed that myometrial contractions induced by peristaltic propulsion and pressure gradient typically regulate the intrauterine fluid movement. Several factors like changes in the mechanical behavior of uterus with age, hormonal variations, number of pregnancies, and mode of delivery (vaginal delivery versus cesarean), and any gestation may impair this mechanism. There are limited reports which indicate that the intrauterine fluid movement is possible to control with the help of external electric or magnetic fields. However, there are very few studies that investigate the intrauterine fluid motion in the presence of aforementioned external stimuli. This encouraged us to develop a computer model to study intrauterine fluid movement induced by peristalsis in the presence of electro-magnetohydrodynamics. Uterus geometry is idealized as a tapered micro-channel, whereas the Williamson fluid model is used to represent the intrauterine fluid. The Debye-Huckel linearization and perturbation method are employed to obtain the mathematical solution for fluid motion in the presence of any external field. The effect of the Debye-Huckel parameter, Helmholtz-Smoluchowski velocity, zeta potential and Hartmann number on pumping characteristics, flow characteristics, shear stress and trapping are studied to analyze the sensitivity of the fluid model. Overall, this study presents a model to understand the behavior of the intrauterine fluid motion in the presence of electric and magnetic fields. The model and associated results may be encouraged and may helpful for biomedical engineers in the design and development of such biomicrofluidic devices which can transport the embryo at a suitable location of the uterus for implantation.

Development of a Method to Characterize Vascular Contributions to Cramping Pain in Dysmenorrhea

Objective: Increased vascular resistance during myometrial contractions could theoretically induce transient ischemia and menstrual cramping pain. However, diagnostic tests have not yet been developed to evaluate vascular contributions to cramping pain in dysmenorrhea. This pilot study used a novel method to examine changes in uterine artery resistance during self-reported cramping.

Repurposing simvastatin as a therapy for preterm labor: evidence from preclinical models.

Preterm birth (PTB), the leading cause of neonatal morbidity and mortality, urgently requires novel therapeutic agents. Spontaneous PTB, resulting from preterm labor, is commonly caused by intrauterine infection/inflammation. Statins are well-established, cholesterol-lowering drugs that can reduce inflammation and inhibit vascular smooth muscle contraction. We show that simvastatin reduced the incidence of PTB in a validated intrauterine LPS-induced PTB mouse model, decreased uterine proinflammatory mRNA concentrations (IL-6, Cxcl1, and Ccl2), and reduced serum IL-6 concentration. In human myometrial cells, simvastatin reduced proinflammatory mediator mRNA and protein expression (IL-6 and IL-8) and increased anti-inflammatory cytokine mRNA expression (IL-10 and IL-13). Critically, simvastatin inhibited myometrial cell contraction, basally and during inflammation, and reduced phosphorylated myosin light chain concentration. Supplementation with mevalonate and geranylgeranyl pyrophosphate, but not farnesyl pyrophosphate, abolished these anticontractile effects, indicating that the Rho/Rho-associated protein kinase pathway is critically involved. Thus, simvastatin reduces PTB incidence in mice, inhibits myometrial contractions, and exhibits key anti-inflammatory effects, providing a rationale for investigation into the repurposing of statins to treat preterm labor in women.—Boyle, A. K., Rinaldi, S. F., Rossi, A. G., Saunders, P. T. K., Norman, J. E. Repurposing simvastatin as a therapy for preterm labor: evidence from preclinical models.

Prostaglandin D2 evokes potent uterine contraction via the F prostanoid receptor in postpartum rats

Prostaglandin (PG) D2, a prostanoid known to have hypotensive effect, can evoke increased in vitro prepartum myometrial contraction resulting from up-regulation of the F prostanoid (FP) receptor. The present study further determined postpartum rat uterine responses to PGD2 to evaluate the possibility of the prostanoid becoming a therapeutic for postpartum uterine atony, a major cause of postpartum hemorrhage that can lead to maternal morbidity. In vitro and in vivo postpartum uterine responses to PGD2 were determined and compared to those of prepartum rats. Here we show that in postpartum myometrial strips PGD2 did evoke a contraction sensitive to FP receptor antagonism. Interestingly, this response was not only to a greater extent than that of prepartum rats, but also comparable with the contraction obtained with PGF2α, a therapeutic for postpartum uterine atony but contradicted in conditions including hypertension. Indeed, PGD2 was also found to cause increases of basal uterine contraction under in vivo conditions. Western blots revealed that the expression of FP receptors in postpartum myometrium was higher than that of prepartum rats. Moreover, we noted that the amount of PGD2 produced in postpartum uteri, although lower than that of prepartum rats, was increased compared to non-pregnant conditions. These results thus demonstrate that due to a further up-regulation or high expression of myometrial FP receptors, PGD2 can evoke potent uterine contraction postpartum, and hence the prostanoid, which is naturally synthesized in uterine tissues, could be a potential therapeutic for postpartum uterine atony, especially in settings, such as hypertension.

Do chlorinated insecticides (aldrin and DDT) or products of their transformations (dieldrin and DDE) impair signal transfer from regulators (oxytocin and relaxin) of bovine myometrium motility in vitro?

Aldrin, dieldrin, and DDT are chlorinated insecticides that are unintentionally widespread in the environment. It was previously shown that all of the aforementioned compounds increased secretion of ovarian oxytocin (OT), which is a potent uterotonic agent. However, only DDT and its metabolite (DDE) promoted, while aldrin and dieldrin inhibited basal and OT-stimulated myometrial contractions in cows. Therefore, the aim of this study was to determine the effect of these treatments on the reception and further transmission of the OT-signal for myometrial contractions and on the levels of contractile-associated integral proteins (caveolin; CAV) and gap junction proteins (GAPs). Moreover, their effect on reception of signal for the relaxation of myometrium was also studied. Myometrial strips or cells from non-pregnant (8–12 days of oestrous cycle) or late pregnant (5–8 months) cows were incubated with the studied compounds at environmentally relevant dose (10 ng/ml), which was chosen according to the previous studies. DDT and DDE increased the CAV protein level, while dieldrin decreased the GAPs level. None of the studied compounds affected mRNA expression of the OT receptor and expression of the second messengers (DAG, IP3, PKC, MLCK). Oppositely, DDE and dieldrin decreased mRNA expression of the relaxin (RLX) receptor. Changes in the amount of contractile-associated integral proteins may be involved in the molecular mechanism underlying the adverse effects of the studied insecticides on myometrial motility. Admittedly, none of the studied compounds impaired the reception or further intracellular transmission of the OT signal to promote contractions during the oestrous cycle, while they showed potential to impair the transmission the signal between cells as well as to diminish the effects of one of the primary inhibitor (RLX) of myometrial contractions during gestation.

The steroid hormone dydrogesterone inhibits myometrial contraction independently of the progesterone/progesterone receptor pathway

AimsIn this study, we aimed to investigate the direct effects of steroid hormones on pregnant myometrial contraction. Main methodsThe effect of steroids on oxytocin-induced contraction was examined in vitro using pregnant rat or human myometrium. Subsequently, we evaluated whether RU486, a potent progesterone antagonist, influenced the effects of progestin on myometrial contraction. Additionally, we evaluated the effects of progestin on high-concentration KCl-induced contraction caused by voltage-dependent calcium channels in order to investigate the mechanisms involved in this process. Key findingsOf the natural steroids examined, 17β-estradiol, progesterone, testosterone, cortisol, and aldosterone did not influence oxytocin-induced contraction at concentrations <10−6 M. Of the tested progestins, medroxyprogesterone acetate, norethisterone, chlormadinone acetate, levonorgesterol, 17α-hydroxyprogesterone capronate, and dienogest had no effect on contraction at <10−6 M. However, dydrogesterone showed rapid and direct inhibition of contraction at 10−6 M, and this inhibitory effect was dependent on dose and time. RU486 did not block the inhibitory effects of dydrogesterone on contraction. High-concentration KCl-induced contraction was also inhibited by dydrogesterone, and the inhibitory effects of dydrogesterone were observed at concentrations as low as 10−7 M. Additionally, oxytocin-induced contraction in pregnant human myometrium was inhibited by 10−6 M dydrogesterone. SignificanceThese results suggested that the rapid and direct effects of dydrogesterone on myometrial contraction were caused by a nongenomic pathway and that the progesterone receptor was not required for dydrogesterone action. Additionally, the mechanism of dydrogesterone action may involve voltage-dependent calcium channels.

Transient uterine contractions as a potential pathology mimic on premenopausal pelvic MRI and the role of routine repeat T2 sagittal images to improve observer confidence.

Pelvic MRI has an increasingly important role in the evaluation of non-malignant uterine pathology including uterine leiomyomas, adenomyosis and endometriosis. Normal physiological myometrial junctional zone transient contractions can also be identified on MRI and have the potential to act as pathology mimics. This study aims to evaluate both the incidence of visible physiological contractions in premenopausal female pelvic MRI and also to support the routine acquisition of a repeat T2 sagittal sequence to differentiate transient physiological contractions from true underlying pathology and therefore improve observer confidence. A total of 279 female patients of child-bearing age who had undergone a pelvic MRI over a 16month period met the inclusion criteria. All patients underwent a standard examination protocol on the same hardware. This included performing two separate T2-weighted sagittal sequences as part of the protocol firstly as the initial and then as the final series for the examination. The sagittal series were reviewed separately by four readers and conclusions made for each case with regards to the presence of identifiable contractions on one or both series and their potential to act as pathology mimics. Of the 279 cases, there were 34 cases (12.2%) that were found to have transient junctional zone contractions acting as pathology mimics, resembling either leiomyomata or adenomyosis. Standard MRI sequences need to be able to distinguish normal transient physiological uterine contractions from true pathology to avoid diagnostic error. The routine utility of a repeat T2-weighted sagittal sequence performed at the conclusion of a patient's examination was shown to improve reader confidence in distinguishing transient contractions from true uterine pathology while adding minimal time penalty to the overall examination. It is therefore advocated that all premenopausal female pelvic MRI cases have a T2 sagittal series as the initial and then the final series as part of a routine protocol.

Abscisic acid: a novel uterine stimulator in normal and diabetic rats.

Diabetes is usually associated with alterations in myometrial contractility with altered oxytocin responsiveness that increase the incidence of fetal and maternal morbidity and mortality. Pancreatic β-cells release abscisic acid (ABA) in response to glucose, which in turn potentiates insulin secretion. The aim of the study was to find out the effect of ABA on the uterine contractility in normal and diabetic induced rats and tried to detect its possible underlying signaling pathway. Adult non-pregnant female rats were divided into normal nondiabetic group (n = 27) and diabetic group (n = 12). The effect of ABA on the normal and diabetic isolated myometrium was determined alone or after different blockers. Spontaneous diabetic myometrial contraction showed significant decrease and less responsiveness to oxytocin, KCL, and acetylcholine than nondiabetic samples. ABA showed 60% of oxytocin stimulatory effects on myometrial contraction in a dose-response manner in both groups. Meanwhile, this effect was decreased after blocking L-type calcium channels and completely abolished after blocking prostaglandin F (FP) and inositol trisphosphate (IP3) receptors. ABA is found to have an uterotonic effect that is mediated mainly via FP receptor through increasing the level of IP3. So, ABA by its novel effect could be beneficial as pre-labor prescription, especially in diabetic females.

Calcium Channels, Rho-Kinase, Protein Kinase-C, and Phospholipase-C Pathways Mediate Mercury Chloride-Induced Myometrial Contractions in Rats.

Adverse effects of mercury on female reproduction are reported; however, its effect on myogenic activity of uterus and mechanism thereof is obscure. Present study was undertaken to unravel the mechanistic pathways of mercuric chloride (HgCl2)-induced myometrial contraction in rats. Isometric tension in myometrial strips of rats following in vitro exposure to HgCl2 was recorded using data acquisition system-based physiograph. HgCl2 produced concentration-dependent (10nM-100μM) uterotonic effect which was significantly (p < 0.05) reduced in Ca2+-free solution and inhibited in the presence of nifedipine (1μM), a L-type Ca2+ channel blocker, thus suggesting the importance of extracellular Ca2+ and its entry through L-type calcium channels in HgCl2-induced myometrial contractions in rats. Cumulative concentration-response curve of HgCl2 was significantly (p < 0.05) shifted towards right in the presence of Y-27632 (10μM), a Rho-kinase inhibitor, suggesting the involvement of Ca2+-sensitization pathway in mediating HgCl2-induced myometrial contraction. HgCl2-induced myometrial contraction was also significantly (p < 0.05) inhibited in the presence of methoctramine or para-fluoro-hexahydro-siladifenidol, a selective M2 and M3 receptor antagonists, respectively, which evidently suggest that mercury also interacts with M2 and M3 muscarinic receptors to produce myometrial contractions. U-73122 and GF-109203X, the respective inhibitors of PLC and PKC-dependent pathways, downstream to the receptor activation, also significantly (p < 0.05) attenuated the uterotonic effect of HgCl2 on rat uterus. Taken together, present study evidently reveals that HgCl2 interacts with muscarinic receptors and activates calcium signaling cascades involving calcium channels, Rho-kinase, protein kinase-C, and phospholipase-C pathways to exert uterotonic effect in rats. Graphical Abstract Graphical abstract depicting the mechanism of mercury-induced myometrial contraction in rats. M receptor: Muscarinic receptor; PIP2: phospho-inositol bisphosphate; PLC: phospholipase-C; DAG: diacyl glycerol; IP3: inositol triphosphate; IP3R: inositol triphosphate receptor; PKC; protein kinase-C; MLCP: myosin light chain phosphatise; MYPT: myosin phosphatase; SR: sarco-endoplasmic reticulum.

OBE022, an Oral and Selective Prostaglandin F2α Receptor Antagonist as an Effective and Safe Modality for the Treatment of Preterm Labor.

Preterm birth is the major challenge in obstetrics, affecting ∼10% of pregnancies. Pan-prostaglandin synthesis inhibitors [nonsteroidal anti-inflammatory drugs (NSAIDs)] prevent preterm labor and prolong pregnancy but raise concerns about fetal renal and cardiovascular safety. We conducted preclinical studies examining the tocolytic effect and fetal safety of the oral prodrug candidate OBE022 [(S)-2-amino-3-methyl-butyric acid (S)-3-{[(S)-3-(biphenyl-4-sulfonyl)-thiazolidine-2-carbonyl]-amino}-3-(4-fluoro-phenyl)-propyl ester] and its parent OBE002 [(S)-3-(biphenyl-4-sulfonyl)-thiazolidine-2-carboxylic acid [(S)-1-(4-fluoro-phenyl)-3-hydroxy-propyl]-amide], both potent and highly selective antagonist of the contractile prostaglandin F2α (PGF2α ) receptor (FP). Efficacy of OBE022 and OBE002, alone and in combination with other tocolytics, was assessed in human tissues and pregnant animal models for inhibition of uterine contraction and delay of parturition. Selective safety of OBE022 and/or OBE002, compared with NSAID indomethacin, was assessed on renal function, closure of the ductus arteriosus, and inhibition of platelet aggregation. In in vitro studies, OBE002 inhibited spontaneous, oxytocin- and PGF2α -induced human myometrial contractions alone and was more effective in combination with atosiban or nifedipine. In in vivo studies, OBE022 and OBE002 reduced spontaneous contractions in near-term pregnant rats. In pregnant mice, OBE022 delayed RU486 [(8S,11R,13S,14S,17S)-11-[4-(dimethylamino)phenyl]-17-hydroxy-13-methyl-17-prop-1-ynyl-1,2,6,7,8,11,12,14,15,16-decahydrocyclopenta[a]phenanthren-3-one] -induced parturition and exerted synergistic effects in combination with nifedipine. OBE022 and/or OBE002 did not show the fetal side effects of ductus arteriosus constriction, impairment of kidney function, or inhibition of platelet aggregation observed with indomethacin. Orally active OBE022 and OBE002 exhibits potent tocolytic effects on human tissues ex vivo and animal models in vivo without causing the adverse fetal side effects seen with indomethacin. Selectively targeting the FP receptor in combination with existing tocolytics may be an effective strategy for preventing or delaying preterm delivery.

Bisphenol A decreases the spontaneous contractions of rat uterus in vitro through a nitrergic mechanism.

Bisphenol A (BPA), a chemical used in the manufacture of plastics, has toxic effects on various systems of the human body including the reproductive system. BPA possesses estrogenic activity and is implicated in altering oogenesis, ovulation, and fertility. In addition to ovulatory changes, uterine contractility is an important factor for fertility. However, the effects of BPA on myometrial contractions are not known. Therefore, we examined the effect of BPA on rat uterine contractions. The uterus was isolated from adult rats showing estrous phase, and spontaneous in vitro contractions were recorded (35±1 °C). The effect of cumulative concentrations of BPA was determined. Further, the involvement of nitric oxide (NO) and guanylyl cyclase (GC) for the BPA-induced changes on uterine contractility was evaluated using the NO synthase inhibitor (L-NAME) or GC inhibitor (methylene blue). BPA decreased the amplitude and frequency of spontaneous uterine contractions in a concentration-dependent manner. A decrease of 50% occurred at 1 and 3 μM for amplitude and frequency, respectively. L-NAME (N-ω-nitro-l-arginine methyl ester) blocked the BPA-induced decrease in amplitude at all concentrations but antagonized the frequency only at the maximum concentration (10 μM). Methylene blue (a GC inhibitor) did not block the BPA-induced responses but for the frequency at 10 μM of BPA. The results indicate that BPA decreased the amplitude and frequency of spontaneous uterine contractions by involving the nitrergic mechanism; however, the GC mechanism is not involved in the depression.

Glyphosate affects the secretion of regulators of uterine contractions in cows while it does not directly impair the motoric function of myometrium in vitro

The effects of pure glyphosate and its most popular product (brand name: Roundup) on the secretion of hormones involved in the regulation of myometrial contractions as well as their direct effects on myometrial contractions were examined. Myometrial strips as well as uterine and ovarian cells were taken from cows during the oestrous cycle and they were treated with both compounds at concentrations from their environmental range. Glyphosate stimulated the secretion of oestradiol from granulosa cells while both herbicides increased and decreased oxytocin (OT) and progesterone secretion from luteal cells respectively. However only Roundup stimulated mRNA expression of the precursor of OT. Both compounds decreased the secretion of prostaglandins from endometrial cells while they exerted no effect on the basal and OT-stimulated force of myometrial contractions. The studied herbicides did not directly impair the motoric function of the myometrium. However our data indicate the potential of these compounds to disturb the secretory functions of the ovaries and uterus which can lead to the deregulation of uterine contractions and to the impairment of fertilisation or to difficulties in the maintenance of gestation.

Maternal obesity-induced decreases in plasma, hepatic and uterine polyunsaturated fatty acids during labour is reversed through improved nutrition at conception

Maternal obesity is associated with prolonged and dysfunctional labour, potentially through decreased synthesis of prostaglandins that stimulate myometrial contractions. We assessed the impact of maternal obesity on concentrations of precursor fatty acids (FA) for prostaglandin synthesis and whether any changes could be reversed by improved nutrition post-conception. Wistar rats were fed control (CON) or High-Fat, High-cholesterol (HFHC) diets 6 weeks before mating. At conception half the dams switched diets providing 4 dietary groups: (1) CON, (2) HFHC, (3) CON-HFHC or (4) HFHC-CON. During parturition rats were euthanized and FA composition of plasma, liver and uterus determined. Visceral fat was doubled in rats exposed to the HFHC diet prior to and/or during pregnancy compared to CON. HFHC diet increased MUFAs but decreased omega-3 and omega-6 PUFAs in plasma and liver. Uterine omega-3 FA concentrations were halved in HFHC versus CON rats, but all other FAs were similar. Switching from HFHC to CON diet at conception restored all FA profiles to those seen in CON rats. The increased MUFA and decreased PUFA concentrations in obese HFHC dams may contribute to aberrant prostaglandin synthesis and dysfunctional myometrial activity and it may be possible to reverse these changes, and potentially improve labour outcomes, by improving nutrition at conception.

Chlorinated insecticides (toxaphene and endrin) affect oxytocin, testosterone, oestradiol and prostaglandin secretion from ovarian and uterine cells as well as myometrial contractions in cow in vitro

We examined the direct effects of toxaphene and endrin, chlorinated insecticides that are widespread in the environment, on myometrial contractions and on the secretion of hormones involved in regulating these contractions. Granulosa, luteal, endometrial and myometrial cells, and myometrial strips from non-pregnant cows were incubated with both insecticides at environmentally relevant doses.Toxaphene inhibited and endrin stimulated the secretion of testosterone and oestradiol from granulosa cells. Toxaphene also inhibited and endrin stimulated the expression of the mRNA encoding the precursor of oxytocin (OT), as well OT secretion in luteal cell cultures. Moreover, endrin increased OT secretion from granulosa cells. Neither insecticide exerted an effect on progesterone secretion from luteal cells. Only toxaphene decreased the secretion of prostaglandins (PGF2 and PGE2) from endometrial cells. Meanwhile, only endrin decreased basal myometrial contractions, which was accompanied by inhibition of PGF2 secretion from the myometrium. Both endrin and toxaphene also decreased the force of the OT-stimulated myometrial contractions, whereas only toxaphene inhibited the stimulatory effect of OT on the force of myometrial contractions.In contrast to endrin, toxaphene decreased synthesis and secretion of one of the primary stimulators of myometrial contractions (OT) and indirectly inhibited OT signal reception in the myometrium by reducing E2 secretion. Both insecticides decreased OT-stimulated myometrial contractions; therefore, they may inhibit further transmission of the OT signal. Moreover, endrin inhibited basal myometrial contractions, potentially resulting from reduced PGF2 secretion from the myometrium. Our data indicate the potential of these insecticides to disturb the course of the oestrous cycle or fertilisation.

Contractility Measurements of Human Uterine Smooth Muscle to Aid Drug Development

Discovery and characterization of novel pharmaceutical compounds or biochemical probes rely on robust and physiologically relevant assay systems. We describe methods to measure ex vivo myometrium contractility. This assay can be used to investigate factors and molecules involved in the modulation of myometrial contraction and to determine their excitatory or inhibitory actions, and hence their therapeutic potential in vivo. Biopsies are obtained from women undergoing cesarean section delivery with informed consent. Fine strips of myometrium are dissected, clipped and attached to a force transducer within 1 mL organ baths superfused with physiological saline solution at 37 °C. Strips develop spontaneous contractions within 2–3 h under set tension and remain stable for many hours (>6 h). Strips can also be stimulated to contract such as by the endogenous hormones, oxytocin and vasopressin, which cause concentration-dependent modulation of contraction frequency, force and duration, to more closely resemble contractions in labor. Hence, the effect of known and novel drug leads can be tested on spontaneous and agonist-induced contractions. This protocol specifically details how this assay can be used to determine the potency of known and novel agents by measuring their effects on various parameters of human myometrial contraction. We use the oxytocin- and V1a receptor antagonists, atosiban and SR49059 as examples of known compounds which inhibit oxytocin- and vasopressin-induced contractions, and demonstrate how this method can be used to complement and validate pharmacological data obtained from cell-based assays to aid drug development. The effects of novel agonists in comparison to oxytocin and vasopressin can also be characterized. Whilst we use the example of the oxytocin/ vasopressin system, this method can also be used to study other receptors and ion channels that play a role in uterine contraction and relaxation to advance the understanding of human uterine physiology and pathophysiology.