Uterine Perfusion Research Articles

S-43-3: INTRAUTERINE GROWTH RESTRICTION CAUSES AN ALTERATION IN KIDNEY GENE EXPRESSION

High blood pressure (BP) and kidney disease are the leading cause of death in the world population. Experimental and epidemiological studies indicate that the fetal environment is critical in determining adult cardiovascular (CV)-renal outcome. It is known that intrauterine growth restriction (IUGR) or low birth weight (LBW) induces an increased risk of hypertension and kidney disease in later life. Placental insufficiency is the common cause of IUGR. Low birth weight is a significant public health problem globally and is associated with short- and long-term consequences. About 15–20% of all births worldwide are LBW, representing more than 20 million births a year. Clinical studies indicate that birth weight is positively associated with glomerular filtration rate (GFR) and negatively associated with BP. Also, animal models of IUGR exhibit a higher susceptibility to ANGII induced high BP and kidney damage in response to acute ischemic reperfusion injury. Thus, these data suggest that IUGR programs greater risk of hypertension and renal dysfunction. However, the exact mechanisms linking an adverse fetal environment with a later increased risk of these diseases are unclear. Objective: This study aims to evaluate the global genetic change in the kidney of the animal model of IUGR. Design: female C57Bl/6J were mated with male C57bl/6J. Upon confirming the pregnancy by sperm plug (day 0 of gestation), the pregnant animal will be undergone reduced uterine perfusion surgery (RUP) or shame surgery at day 13 of gestation. The offspring of RUP mom will be labeled as IUGR, and from the sham will be labeled as control. The offspring will be used for blood pressure measurement using radio-telemetry and kidney microarray analysis when they reach 24 weeks. Results: Th IUGR offspring significantly has an increase in blood pressure compared to the control. The kidneys of male IUGR have 6529 gene upregulation compared to male control. The make IUGR also has 5713 genes dow-regulation as compared to same-sex control. The female IUGR has 1635 genes upregulated and 1513 genes downregulated compared to the kidney from the female control group. Conclusion: the complication during pregnancy, such as reduced uterine perfusion, which is commonly found in preeclampsia programs, alters gene expression in the kidneys of their offspring. This finding suggests that IUGR causes the gene alteration in the kidneys, which is the primary organ in controlling the blood pressure of the offspring, and may lead to the development of hypertension and kidney disease.

AMPK regulates homeostasis of invasion and viability in trophoblasts by redirecting glucose metabolism: Implications for pre‐eclampsia

Pre-eclampsia (PE) is deemed an ischemia-induced metabolic disorder of the placenta due to defective invasion of trophoblasts during placentation; thus, the driving role of metabolism in PE pathogenesis is largely ignored. Since trophoblasts undergo substantial glycolysis, this study aimed to investigate its function and regulatory mechanism by AMPK in PE development. Metabolomics analysis of PE placentas was performed by gas chromatography-mass spectrometry (GC-MS). Trophoblast-specific AMPKα1-deficient mouse placentas were generated to assess morphology. A mouse PE model was established by Reduced Uterine Perfusion Pressure, and placental AMPK was modulated by nanoparticle-delivered A769662. Trophoblast glucose uptake was measured by 2-NBDG and 2-deoxy-d-[3 H] glucose uptake assays. Cellular metabolism was investigated by the Seahorse assay and GC-MS.PE complicated trophoblasts are associated with AMPK hyperactivation due not to energy deficiency. Thereafter, AMPK activation during placentation exacerbated PE manifestations but alleviated cell death in the placenta. AMPK activation in trophoblasts contributed to GLUT3 translocation and subsequent glucose metabolism, which were redirected into gluconeogenesis, resulting in deposition of glycogen and accumulation of phosphoenolpyruvate; the latter enhanced viability but compromised trophoblast invasion. However, ablation of AMPK in the mouse placenta resulted in decreased glycogen deposition and structural malformation. These data reveal a novel homeostasis between invasiveness and viability in trophoblasts, which is mechanistically relevant for switching between the 'go' and 'grow' cellular programs.

Decreased expression of caveolin-1 have relevance to promoted senescence in preeclamptic placenta.

To assess the association between altered expression of caveolin-1 and p53/p21, as indicatives of cellular senescence, in preeclamptic placenta. Placental tissues and serum were collected from rats (Sham and reduced uterine perfusion pressure group) at 18.5days post coitum and humans (normotensive pregnant and preeclampsia groups). The concentration and expression of caveolin-1 were measured in the collected tissues, and the correlation between p53 and p21 expression was evaluation. Placental mRNA expression and serum concentration of caveolin-1 were measured using qRT-PCR and ELISA, respectively. Altered expressions of caveolin-1 and p53/p21 were revealed and quantified by immunohistochemistry. The association between these changes was investigated using correlation analysis. Placental mRNA expressions and serum concentrations of caveolin-1 were significantly decreased in reduced uterine perfusion pressure and preeclampsia groups. The expressions of caveolin-1 and p53/ p21 were significantly altered in placenta complicated with preeclampsia. Correlation analysis revealed a significant inverse association between changes in caveolin-1 and p53/p21. Subsequently, these results were obtained by investigating the preeclampsia onset time. These results revealed that the expression of caveolin-1 profoundly decreases in the placenta and serum of preeclampsia. These factors contribute to the mechanism of accelerated cellular senescence in placenta, which is one of the various etiologies of preeclampsia.

Increased Ca2+-dependent intrinsic tone and arterial stiffness in mesenteric microvessels of hypertensive pregnant rats

Preeclampsia is a pregnancy-related hypertensive disorder (HTN-Preg) with unclear mechanisms. We have shown increased vascular reactivity to extrinsic vasoconstrictors in HTN-Preg rats. Here, we test whether microvascular intrinsic tone and arterial stiffness could contribute to HTN-Preg, and examined the underlying cellular mechanisms. On gestational day 19, BP was recorded in normal pregnant (Preg) rats and Preg rats with reduced uterine perfusion pressure (RUPP), and mesenteric microvessels were mounted on a pressure myograph for measurement of intrinsic tone, simultaneous changes in [Ca2+]i (fura-2 340/380 ratio), and arterial stiffness. Arteries were incubated in Ca2+-containing and 0 Ca2+ (2 mM EGTA) Krebs, pressurized at 10 to 110 mmHg in 10 mmHg increments, and the % change in vessel diameter from initial diameter at 10 mmHg was analyzed for measurement of total (active + passive) intrinsic tone and passive intrinsic response, respectively. The passive response was then subtracted from the total intrinsic tone to determine the active myogenic tone. The strain–stress relationship was also constructed as a measure of arterial stiffness. BP was higher in RUPP vs Preg rats. In Ca2+-containing Krebs, increases in intraluminal pressure caused smaller increases in diameter and greater increases in [Ca2+]i in microvessels of RUPP vs Preg rats, suggesting increased Ca2+-dependent myogenic tone. In 0 Ca2+ Krebs, increases in pressure also caused less increases in diameter in microvessels of RUPP vs Preg rats, but with no changes in [Ca2+]i, suggesting changes in the structure and mechanics of the arterial wall. The total and passive strain–stress relationship was shifted to the left in microvessels of RUPP vs Preg rats, suggesting increased arterial wall stiffness. Histology and immunohistochemistry showed greater vascular wall thickness and collagen-I staining in RUPP vs Preg rats, supporting changes in the wall architecture and structural proteins. The increased active myogenic tone and underlying increases in Ca2+ signaling as well as the increased passive intrinsic response, arterial stiffness and collagen-I in the mesenteric microvessels could play a role in the regulation of blood flow to the splanchnic region and the increased vascular resistance and BP in HTN-Preg.

MiR-2115-3p inhibits ferroptosis by downregulating the expression of glutamic-oxaloacetic transaminase in preeclampsia

IntroductionRecently, ferroptosis has been reported to be closely related to preeclampsia (PE). However, the mechanisms associated with ferroptosis in PE have been poorly studied. MethodsA PE rat model was established via reduced uterine perfusion pressure (RUPP) surgery. A Ferroptosis inhibitor was used to treat the model rats. Blood pressure, urine protein, sFlt-1, GSH, GSH-PX, MDA, total Fe, Fe (II), and Fe (Ⅲ) levels were detected. Placenta morphological changes and fetal survival rate were observed and counted, respectively. miRNA sequencing and bioinformatical analysis were conducted to establish the ferroptosis-related interaction network of miRNAs-mRNAs in PE. After hypoxia treatment, cells were silenced glutamic-oxaloacetic transaminase1 (GOT1) or overexpressed miR-2115-3p/GOT1. Cellular proliferation, invasion, and reactive oxygen species (ROS) were determined. The ferroptosis-related factors levels (ACSL4, TfR1, GPX4, SCL7A11, GSH, GSH-PX, Fe (II), and MDA) were quantified. ResultsIn vivo, ferrostatin-1 attenuated ferroptosis in the PE models, significantly increasing fetal survivability. The miR-2115-3p/GOT1 pathway was screened for possible association with abnormal ferroptosis in PE. miR-2115-3p was discovered to interact with the mRNA of GOT1. Inhibition of GOT1 and overexpression of miR-2115-3p reversed the decrease in cell proliferation capacity, GSH, GSH-PX, and GPX4 levels, and the increase in ROS, ACSL4, TfR1, MDA, total Fe, and Fe (II) levels induced by hypoxia. However, simultaneous overexpression of miR-2115-3p and GOT1 reversed the above results of overexpression of miR-2115-3p. DiscussionmiR-2115-3p might interact with the GOT1 mRNA to downregulate its expression, further inhibiting the hypoxia-promoted ferroptosis in a PE model.

Hormone therapy for uterine and endometrial development in women with premature ovarian insufficiency.

Premature ovarian insufficiency (POI) is a clinical syndrome resulting from loss of ovarian function before the age of 40. It is a state of hypergonadotropic hypogonadism, characterised by amenorrhoea or oligomenorrhoea, with low ovarian sex hormones (oestrogen deficiency) and elevated pituitary gonadotrophins. POI with primary amenorrhoea may occur as a result of chromosomal and genetic abnormalities, such as Turner syndrome, Fragile X, or autosomal gene defects; secondary amenorrhoea may be iatrogenic after the surgical removal of the ovaries, radiotherapy, or chemotherapy. Other causes include autoimmune diseases, viral infections, and environmental factors; in most cases, POI is idiopathic. Appropriate replacement of sex hormones in women with POI may facilitate the achievement of near normal uterine development. However, the optimal effective hormone therapy (HT) regimen to maximise the reproductive potential for women with POI remains unclear. To investigate the effectiveness and safety of different hormonal regimens on uterine and endometrial development in women with POI. We searched the Cochrane Gynaecology and Fertility (CGF) Group trials register, CENTRAL, MEDLINE, Embase, PsycINFO, CINAHL, and two trials registers in September 2021. We also checked references of included studies, and contacted study authors to identify additional studies. We included randomised controlled trials (RCTs) investigating the effect of various hormonal preparations on the uterine development of women diagnosed with POI. We used standard methodological procedures recommended by Cochrane. The primary review outcome was uterine volume; secondary outcomes were endometrial thickness, endometrial histology, uterine perfusion, reproductive outcomes, and any reported adverse events. We included three studies (52 participants analysed in total) investigating the role of various hormonal preparations in three different contexts, which deemed meta-analysis unfeasible. We found very low-certainty evidence; the main limitation was very serious imprecision due to small sample size. Conjugated oral oestrogens versus transdermal 17ß-oestradiol We are uncertain of the effect of conjugated oral oestrogens compared to transdermal 17ß-oestradiol (mean difference (MD) -18.2 (mL), 95% confidence interval (CI) -23.18 to -13.22; 1 RCT, N = 12; very low-certainty evidence) on uterine volume, measured after 12 months of treatment. The study reported no other relevant outcomes (including adverse events). Low versus high 17ß-oestradiol dose We are uncertain of the effect of a lower dose of 17ß-oestradiol compared to a higher dose of 17ß-oestradiol on uterine volume after three or five years of treatment, or adverse events (1 RCT, N = 20; very low-certainty evidence). The study reported no other relevant outcomes. Oral versus vaginal administration of oestradiol and dydrogesterone We are uncertain of the effect of an oral or vaginal administration route on uterine volume and endometrial thickness after 14 or 21 days of administration (1 RCT, N = 20; very low-certainty evidence). The study reported no other relevant outcomes (including adverse events). No clear conclusions can be drawn in this systematic review, due to the very low-certainty of the evidence. There is a need for pragmatic, well designed, randomised controlled trials, with adequate power to detect differences between various HT regimens on uterine growth, endometrial development, and pregnancy outcomes following the transfer of donated gametes or embryos in women diagnosed with POI.

Inhibition of the AT1R agonistic autoantibody in a rat model of preeclampsia improves fetal growth in late gestation.

Placenta ischemia, the initiating event in preeclampsia (PE), is associated with fetal growth restriction. Inhibition of the agonistic autoantibody against the angiotensin type 1 receptor AT1-AA, using an epitope-binding inhibitory peptide ('n7AAc') attenuates increased blood pressure at gestational day (G)19 in the clinically relevant reduced uterine perfusion pressure (RUPP) model of PE. Thus we tested the hypothesis that maternal administration of 'n7AAc' does not transfer to the fetus, improves uterine blood flow and fetal growth, and attenuates elevated placental expression of miRNAs implicated in PE and FGR. Sham or RUPP surgery was performed at G14 with vehicle or 'n7AAc' (144 µg/day) administered via an osmotic pump from G14 to G20. Maternal plasma levels of the peptide on G20 were 16.28 ± 4.4 nM, and fetal plasma levels were significantly lower at 1.15 ± 1.7 nM (P = 0.0007). The uterine artery resistance index was significantly elevated in RUPP (P < 0.0001) but was not increased in 'n7AAc'-RUPP or 'n7AAc'-Sham versus Sham. A significant reduction in fetal weight at G20 in RUPP (P = 0.003) was not observed in 'n7AAc'-RUPP. Yet, percent survival was reduced in RUPP (P = 0.0007) and 'n7AAc'-RUPP (P < 0.0002). Correlation analysis indicated the reduction in percent survival during gestation was specific to the RUPP (r = 0.5342, P = 0.043) and independent of 'n7AAc'. Placental miR-155 (P = 0.0091) and miR-181a (P = 0.0384) expression was upregulated in RUPP at G20 but was not elevated in 'n7AAc'-RUPP. Collectively, our results suggest that maternal administration of 'n7AAc' does not alter fetal growth in the RUPP implicating its potential as a therapeutic for the treatment of PE.NEW & NOTEWORTHY The seven amino acid inhibitory peptide to the AT1-AA ('n7AAc') has limited transfer to the fetus at gestational day 20, improves uterine blood flow and fetal growth in the reduced uterine perfusion pressure model of preeclampsia (PE), and does not impair fetal survival during gestation in sham-operated or placental ischemic rats. Collectively, these findings suggest that maternal administration of 'n7AAc' as an effective strategy for the treatment of PE is associated with improved outcomes in the fetus.

The vaginal and uterine blood flow changes during the ovsynch program and its impact on the pregnancy rates in Holstein dairy cows

AimOvSynch is a hormonal protocol for synchronization of estrus and use of artificial insemination (AI) at an optimal time without adverse effects on the ovaries or uterus. This study investigated the use of noninvasive color Doppler ultrasound to assess changes in uterine and vaginal blood flow during the Ovsynch program for synchronization of estrus and its relation to the pregnancy rates in Holstein cows.Materials and methodsThe experimental cows received an intramuscular dose of 10 μg of a GnRH analogue (G1), followed 7 days later with an intramuscular injection of synthetic prostaglandin F2α (P: PGF2α) analogue (500 μg cloprostenol sodium), and given a 10 μg, injection of the GnRH analogue (G2) i.m. 48 h after the PGF2α treatment, and the cows were bred 14-16 h after. Uterine and vaginal perfusion were investigated by performing transrectal Doppler ultrasonography of both the uterine and vaginal arteries in Holstein cows at different time points during the Ovsynch program to determine: peak systolic velocity (PSV), time-averaged maximum velocity (TAMV), the volume of blood flow (BFV), pulsatility index (PI), resistance index (RI), resistance impedance (S/D) and diameters of uterine (UA) and vaginal (VA) arteries. Steroid hormones were also assayed. Transrectal ultrasonography (TUS) was performed at 32 and 60 days to confirm the pregnancy per artificial insemination (P/AI).ResultsThe uterine PSV, TAMV, and PV were greater at the time of the cloprostenol sodium and second GnRH injections (p<0.05) than at the time of the first GnRH injection. The vaginal PSV, PV were greater at the time of the cloprostenol sodium than at the time of the first and second GnRH injections (p<0.05). The receiver operating characteristic curve (ROC curve) indicated a high correlation between the uterine and vaginal blood flow and the rate of the pregnancy (p<0.05). The area under the ROC curve was 0.920 and 0.87 (p<0.05) for vaginal and uterine arteries respectively at time of G2. The serum levels of progesterone, estrogen and cortisol were correlated with the P/AI (p<0.05). The P/AI significantly decreased from 43.9 % at 32 d to 35.37 % at 60 d.ConclusionThese results indicate that noninvasive Doppler ultrasonography is a valid method to evaluate changes in the characteristics of uterine and vaginal blood flow in cows during the Ovsynch protocol. Furthermore, vaginal and uterine blood flow are two determinant factors for the higher conception rates in Holstein dairy cows.

Abstract P349: Decreased Complement Component C3 In Islets Is Associated With A Reduction In Pancreatic Beta Cell Area In Rat Offspring Following Chronic Placental Ischemia-induced Hypertension

Gestational hypertension with or without low birth weight is associated with increased risk for Type 2 diabetes in offspring. Pancreatic beta cell mass is set very early in life and can influence whether an individual develops Type 2 diabetes. We previously demonstrated that postnatal day (PD)13 female offspring of rats subjected to chronic placental ischemia-induced hypertension had significantly reduced beta cell area. Previous studies in humans have noted increased circulating C3 in Type 2 diabetics, as well as an important role for intracellular complement C3 in maintaining integrity of the beta cell in the islet. Thus, we hypothesized that decreased C3 in islets is associated with reductions in beta cell area in rat offspring following placental insufficiency. The rat Reduced Uterine Perfusion Pressure (RUPP) model was used to mechanically induce placental insufficiency in the dam by placing silver clips on abdominal aorta and uterine arteries on gestation day 14 of a 21-day gestation, resulting in hypertension in the dam. Pancreatic islets and acinar were isolated by collagenase perfusion and hand picking from PD13 female offspring of RUPP and Sham dams. The concentration of C3 in serum, isolated islets and acinar was determined by ELISA. C3 was significantly increased in serum of female RUPP compared to female Sham offspring. In contrast, C3 in islets was significantly decreased in RUPP female offspring compared to Sham with no significant differences detected in acinar C3 concentrations. Thus, these data suggest that decreased C3 in islets of PD13 offspring may contribute to reduction of beta cell area and long-term susceptibility for Type 2 diabetes.

Abstract 103: Il-33 Supplementation Improves Vascular Function And Maternal Hypertension In Response To Placental Ischemia

Preeclampsia (PE), a leading cause of maternal and fetal morbidity and mortality, is a hypertensive pregnancy disorder with end-organ damage that develops after 20 weeks of gestation. PE is characterized by chronic immune activation, endothelial dysfunction, and fetal growth restriction (FGR). Clinical studies have shown decreased IL-33 signaling in PE women. We use the Reduced Uterine Perfusion Pressure (RUPP) rat model, which mimics many characteristics of PE patients including reduced IL-33 levels, to identify the mechanisms mediating maternal hypertension and FGR in PE. We hypothesized that IL-33 supplementation would improve maternal hypertension, fetal growth, and oxidative stress (ROS) during placental ischemia in pregnant rats. We implanted intraperitoneal mini-osmotic pumps infusing recombinant rat IL-33 (1 μg/kg/day) in normal pregnant (NP) and RUPP rats from gestation day 14-19. Maternal blood pressure, fetal growth, placental and renal ROS, phosphorylated endothelial nitric-oxide synthase ( p eNOS), and uterine artery resistance index (UARI) were assessed. MAP (mmHg) increased from 104±4 in NP to 124±7 in RUPP (p&lt;0.05) and was significantly lowered in RUPP+IL-33 to 108±10 (p&lt;0.05, n=6-8/group). Fetal weight was significantly increased in RUPP+ IL-33 compared to RUPP (p&lt;0.05). Placental ROS was not different among any groups. However, renal ROS (relative light units/min/mg protein) was increased from 452±33 in NP to 676±44 in RUPP (p&lt;0.05) and was significantly decreased to 309±61 in RUPP+IL-33 (n=4-5/group, p&lt;0.05 vs RUPP). Placental p eNOS expression was 4-fold lower in RUPP vs NP (p&lt;0.05) and 5-fold lower in RUPP+IL-33 vs NP, n=3-5/group). UARI (arbitrary units) was 0.709±0.012 in RUPP vs NP (0.585±0.025, p&lt;0.05) and was significantly lowered to 0.577±0.025 in RUPP+IL-33 rats (p&lt;0.05 vs RUPP, n=5-6/group). These findings demonstrate a role for IL-33 in controlling vascular function and maternal blood pressure during pregnancy independent of eNOS activation. Further studies will quantify effector immune cells and inflammatory cytokine levels in response to IL-33 supplementation during placental ischemia. These data suggest that IL-33 may have therapeutic potential in management of PE pathophysiology during pregnancy.

Abstract 050: Endothelial Cell Mineralocorticoid Receptor Deletion Improves Endothelial Function, Blood Pressure And Fetal Growth In The RUPP Mouse Model Of Preeclampsia

Preeclampsia (PE) is a hypertensive disorder of pregnancy that causes adverse pregnancy outcomes. Progesterone in pregnancy increases endothelial mineralocorticoid receptor (ECMR) expression, which we showed mediates endothelial function in hypertensive female mice, however, whether ablating endothelial dysfunction in PE improves pregnancy outcomes is unknown. We hypothesized that improving endothelial function via ECMR deletion increases fetal growth and ablates hypertension in the reduced uterine perfusion pressure (RUPP) PE mouse. Pregnant ECMR-intact (WT) and -deficient (KO) mice underwent RUPP or sham surgery on GD13 and sacrificed on GD18 (WT Sham=9, WT RUPP=5, KO Sham=6, KO RUPP=6). Vascular function was measured via wire myography on 2 nd order mesenteric arteries and thoracic aorta. Systolic blood pressure (BP) was measured by radiotelemetry in a subset of mice. RUPP did not decrease maternal weights in either WT or KO mice (P&gt;0.05) but decreased pup weight (726±20 WT+Sham vs 660±11mg WT+RUPP, *P&lt;0.05) and fetal demise (16±4 vs 28±4% resoprtions/litter, *P&lt;0.05) in WT pregnant mice, however, not in KO mice (710±15 KO+Sham vs 693±19 mg KO+RUPP, P&gt;0.05, and 13±5 vs 28±12% resoprtions/litter, P&gt;0.05), indicating KO protected fetal growth. RUPP reduced endothelial-dependent, acetylcholine-mediated vascular relaxation (10 -9 -3x10 -5 M) in WT, but not KO pregnant mice (*P&lt;0.05, 2-way ANOVA w/ RM), but not aorta ACh-mediated relaxation (P&gt;0.05). Nitric oxide synthase (NOS) inhibitor LNAME ablated differences in ACh-mediated relaxation between WT Sham and WT RUPP (P&gt;0.05), with no differences observed in KO Sham or KO RUPP, in both aorta and mesenteric arteries, indicating RUPP impaired endothelial function via decreasing NOS activity. Preliminary data demonstrates RUPP increased BP on GD15-17 in WT (127±21 mmHg, N=3), however, not in KO pregnant mice (111±4, N=2). RUPP also ablated circadian rhythm in WT, but not KO, pregnant mice from GD15-18. Collectively, these data indicate that ECMR activation induces reduced pup weight, endothelial dysfunction and hypertension in the RUPP model which may lead to clinical studies for consideration of specific MR antagonists as therapeutic options for endothelial dysfunction in PE pregnancies.

Abnormal cerebral microvascular perfusion and reactivity in female offspring of reduced uterine perfusion pressure (RUPP) mice model

Children born from women with preeclampsia have alterations in cerebral neurovascular development and a high risk for developing cognitive alterations. Because cerebral blood vessels are critical components in cerebrovascular development, we evaluated the brain microvascular perfusion and microvascular reactivity (exposed to external stimuli of warm and cold) in pups born to preeclampsia-like syndrome based on the reduction of uterine perfusion (RUPP). Also, we evaluate the angiogenic proteomic profile in those brains. Pregnant mice showed a reduction in uterine flow after RUPP surgery (−40 to 50%) associated with unfavorable perinatal results compared to sham mice. Furthermore, offspring of the RUPP mice exhibited reduced brain microvascular perfusion at postnatal day 5 (P5) compared with offspring from sham mice. This reduction was preferentially observed in females. Also, brain microvascular reactivity to external stimuli (warm and cold) was reduced in pups of RUPP mice. Furthermore, a differential expression of the angiogenic profile associated with inflammation, extrinsic apoptotic, cancer, and cellular senescence processes as the primary signaling impaired process was found in the brains of RUPP-offspring. Then, offspring (P5) from preeclampsia-like syndrome exhibit impaired brain perfusion and microvascular reactivity, particularly in female mice, associated with differential expression of angiogenic proteins in the brain tissue.

The Performance of the Uterine Artery Resistance Index following Stimulation of the Cervix in the Diagnosis of Endometriosis: A Prospective Study

Background: We speculated that stimulating the uterine cervix of patients may increase blood flow resistance of the uterine artery more significantly in women with endometriosis than those without. Accordingly, the resistance index (RI) of the uterine artery following stimulation of the cervix may offer a novel and promising non-invasive method for diagnosing endometriosis. In this prospective study, the aim was to determine the performance of the uterine artery resistance index (RI) following stimulation of the cervix in the diagnosis of endometriosis. Methods: This prospective cohort study included 270 patients with non-uterine diseases who were scheduled for laparoscopic surgery. Each patient underwent Doppler sonography for peak systolic velocity (PSV), end diastolic velocity (EDV), and RI before and after stimulation of the cervix, in addition to conventional ultrasound examination one day prior to laparoscopic surgery. The cervix was stimulated using a double convex array electronic vaginal probe for real-time tissue elastography. Results: Laparoscopic surgical evaluation revealed endometriosis in 91 (33.7%) patients. Following stimulation of the cervix, the RI of the uterine artery was significantly increased in the endometriosis group (p = 0.00), but no significant increase in the RI was observed in the control group (p = 0.45). The sensitivity and specificity of the post-stimulation RI for diagnosing endometriosis (using a cut off of 0.83) were 85.71% (95% confidence interval (CI): 76.45–91.88) and 88.83% (95% CI: 83.05–92.87), respectively. For peritoneal endometriosis (PEM), sensitivity and specificity were 86.96% (95% CI: 65.33–96.57) and 88.83% (95% CI: 83.05–92.87), respectively. The sensitivity and specificity of transvaginal ultrasonography (TVU) for diagnosing endometriosis were 63.74% (95% CI: 52.93–73.37) and 94.41% (95% CI: 89.68–97.14), respectively. The positive rate for TVU in diagnosing all endometrioses was 63.7%, and this was lower than that for the post-stimulation RI (84.6%, p = 0.001). Conclusions: Stimulation of the cervix leads to a significant decrease in uterine blood perfusion in women with endometriosis. TVU is a sensitive and specific method for the diagnosis of endometrioma, while the RI is a more sensitive parameter for diagnosing PEM than TVU. For the diagnosis of endometriosis, especially for PEM, the post-stimulation RI may prove to be a promising non-invasive diagnostic technique.

Exogenous L-arginine administration improves uterine vascular perfusion, uteroplacental thickness, steroid concentrations and nitric oxide levels in pregnant buffaloes under subtropical conditions.

Heat stress (HS) during pregnancy adversely affects uterine vascular perfusion and foetal development. L-arginine (L-Arg), a nitric oxide (NO) precursor, has been proven to enhance an organ's vascular perfusion. Therefore, this study aimed to examine the effect of L-Arg administration on the pregnant buffaloes' uterine haemodynamics and uteroplacental thickness under environmental HS conditions. For this purpose, pluriparous (n=12) HS-pregnant buffaloes (mid-gestation, 180-190 days; temperature humidity index >85) were haphazardly assigned into two groups, either administered a single intravenous (IV) bolus of 5mg/kg BW of L-Arg Hcl (n=6; ARG) or received normal saline (0.9%; IV; 25 ml) that served as a control group (n=6; CON). Combined uteroplacental thickness (CUPT, mm), middle uterine arteries' (MUA) diameter (mm) and uterine Doppler haemodynamics (ipsilateral and contralateral to the pregnant horn) were examined utilizing B-mode and colour Doppler ultrasonography, respectively. The serum levels of progesterone (P4), estradiol 17β (E2), nitric oxide (NO) and total antioxidant capacity (TAC) were assayed using commercial kits. The experimental time points were - 1, 0, 2, 4, 24, 48, 72, 96 and 120 h post-L-Arg administration. The ARG group showed higher (p < .05) CUPT and MUA diameter values starting at 24 h post-treatment and onward than the control buffalo cows. In addition, there were improvements (p < .05) in the blood flow parameters in the ipsilateral MUA after L-Arg treatment evidenced by lower values of pulsatility and resistive indices starting at 48 and 4h, respectively and onward, and higher values of peak systolic velocity, coloured areas towards the pregnant uterine horn (both 24-72 h). Increases in the NO levels were found during the period between 4 and 120 h in the ARG group compared to the CON group. Moreover, significant increases in the E2 and P4 means were noted in the ARG group, especially at 24 h onward, respectively compared to the CON group. Concerning the TAC status, neither the administration nor the hours affected serum TAC levels in the HS pregnant buffaloes (CON and ARG). In conclusion, L-Arg administration improved uteroplacental thickness and enhanced uterine haemodynamics, NO levels, and steroids production in mid-pregnant buffalo cows under environmental heat stress conditions which could improve foetal growth and development.

Effect of pentaerythritol tetranitrate (PETN) on the development of fetal growth restriction in pregnancies with impaired uteroplacental perfusion at midgestation—a randomized trial

Fetal growth restriction is strongly associated with impaired placentation and abnormal uteroplacental blood flow. Nitric oxide donors such as pentaerythritol tetranitrate are strong vasodilators and protect the endothelium. Recently, we demonstrated in a randomized controlled pilot study a 38% relative risk reduction for the development of fetal growth restriction or perinatal death following administration of pentaerythritol tetranitrate to pregnant women at risk, identified by impaired uterine perfusion at midgestation. Results of this monocenter study prompted the hypothesis that pentaerythritol tetranitrate might have an effect in pregnancies with compromised placental function as a secondary prophylaxis. This study aimed to test the hypothesis that the nitric oxide donor pentaerythritol tetranitrate reduces fetal growth restriction and perinatal death in pregnant women with impaired placental perfusion at midgestation in a multicenter trial. In this multicenter, randomized, double-blind, placebo-controlled trial, 2 parallel groups of pregnant women presenting with a mean uterine artery pulsatility index >95th percentile at 19+0 to 22+6 weeks of gestation were randomized to 50-mg Pentalong or placebo twice daily. Participants were assigned to high- or low-risk groups according to their medical history before randomization was performed block-wise with a fixed block length stratified by center and risk group. The primary efficacy endpoint was the composite outcome of perinatal death or development of fetal growth restriction. Secondary endpoints were neonatal and maternal outcome parameters. Between August 2017 and March 2020, 317 participants were included in the study and 307 were analyzed. The cumulative incidence of the primary outcome was 41.1% in the pentaerythritol tetranitrate group and 45.5% in the placebo group (unadjusted relative risk, 0.90; 95% confidence interval, 0.69-1.17; adjusted relative risk, 0.90; 95% confidence interval, 0.69-1.17; P=.43). Secondary outcomes such as preterm birth (unadjusted relative risk, 0.73; 95% confidence interval, 0.56-0.94; adjusted relative risk, 0.73; 95% confidence interval, 0.56-0.94; P=.01) and pregnancy-induced hypertension (unadjusted relative risk, 0.65; 95% confidence interval, 0.46-0.93; adjusted relative risk, 0.65; 95% confidence interval, 0.46-0.92; P=0.01) were reduced. Our study failed to show an impact of pentaerythritol tetranitrate on the development of fetal growth restriction and perinatal death in pregnant women with impaired uterine perfusion at midgestation. Pentaerythritol tetranitrate significantly reduced secondary outcome parameters such as the incidence of preterm birth and pregnancy-induced hypertension in these pregnancies.

O-026 RCT of INTRA - Uterine administration of granulocyte colony-stimulating factor (G-CSF) before embryo-transfer in resistant thin endometrium in IVF-ICSI cycles

Abstract Study question Does intrauterine administration of G-CSF (granulocyte-colony-stimulating factor) prior to Embryo-Transfer in patients with resistant thin endometrium improve endometrial thickness and pregnancy rate in IVF-ICSI cycles? Summary answer Yes, Intrauterine administration of G-CSF(granulocyte colony-stimulating factor) before embryo transfer in resistant thin endometrium increases endometrial thickness and improves the pregnancy-rate in IVF-ICSI cycles. What is known already Optimal endometrial thickness reflects adequate maturation, which is a key factor for implantation. Proliferative and secretory changes at the endometrial lining are the result of a complex intrauterine environment where sex steroid hormones and local factors play important role. Lucena et al. found that the uterine-infusion of G-CSF increased endometrial thickness resulting in a successful pregnancy . Li et al. evaluated the effectiveness of G-CSF in a thin endometrium in frozen ET-cycles. The implantation and pregnancy rates were significantly higher (Scarpellini &amp; Sbracia 2012,et al.2015), whereas did not affect endometrial thickness, implantation, or pregnancy rates (Barad et al. 2014, Li et al. 2014). Study design, size, duration 86 Infertile patients with thin endometrium below 45 years age (2018 to 2021) were included in this study, in whom traditional treatment with estradiol, sildenafil etc had been unsuccessful. Patients were randomly divided into two groups using computer generated list after proper consent .The study group received intrauterine- G-CSF and control group received placebo (saline) before Embryo Transfer in IVF-ICSI cycles. Primary Outcome Endometrial-thickness, Implantation &amp; Pregnancy rates. Secondary-outcome Abortion-rate Participants/materials, setting, methods The study group (n = 43) received intrauterine infusion of 300 microgramme/1ml of G-CSF, and the control group (n = 43) underwent placebo-saline infusion before Embryo-Transfer. G-CSF was administered by an intrauterine catheter by slow instillation on the day of hCG administration. If the endometrium had not reached at least a 7-mm within 48 h, a second dose was administered following oocyte retrieval. Main results and the role of chance The endometrial growth was significantly different in the two groups, with an improvement in the G-CSF group. Endometrial expansion to Optimal thickness occurred within approximately 48 -72 hour after G-CSF instillation. In all subjects, at the time of administration of G-CSF, endometrial thickness was 6.43 ± 1.49 mm, and,after G-CSF administration, it increased significantly to 8.11 ± 1.42 mm. The IR and PR were statistically significantly higher in the group receiving intrauterine G-CSF (22% and 33%, respectively) as compared to the control group (08% and 14%, respectively). Abortion Rates were comparable in both the groups and difference was not significant. Endometrial instillation of G-CSF may be effective in expanding chronically unresponsive thin endometrium, resistant to conventional remedies. This treatment needs more multicentric trials to assess its potential in improving the implantation chances in IVF-ICSI cycles, thereby improving pregnancy rates. Uterine perfusion or SC injection with G-CSF represents a promising new tool for intractable problem of thin endometrium . Proliferative and secretory changes in the endometrial lining are the result of a complex intrauterine environment where sex steroid hormones and different local factors play an important role for endometrial thickening. Limitations, reasons for caution Our study is not without limitations. We did not have a large number of patients. The main strengths of study were its RCT design to evaluate the efficacy of intrauterine G-CSF administration on IVF success. Further more RCT are required to establish the best route and dose of GCSF administration in thin-endometrium. Wider implications of the findings Several methods have been proposed, to increase endometrial-thickness in women undergoing IVF-ICSI . These therapies included -tocopherol, pentoxifylline, low-dose aspirin, sildenafil citrate, estradiol administration and hCG priming.Our study using Intra-Uterine instillation of G-CSF represents a promising and cost-effective therapy for the currently most intractable problem of resistant, thin endometrium. Trial registration number not applicable

Coilin as a regulator of NF-kB mediated inflammation in preeclampsia.

ABSTRACTThe nuclear factor-Kappa B (NF-κB) pathway is a crucial mediator of inflammatory signaling. Aberrant activation of NF-κB is associated with several disorders including preeclampsia (PE). Many regulators of the NF-κB pathway have been identified, including microRNAs (miRNAs). Specifically, miR-517-3p targets mRNA encoding TNFAIP3 Interacting Protein 1 (TNIP1), an inhibitor of NF-κB signaling. Activation of NF-κB increases production of the cytokine TNF superfamily member 15 (TNFSF15), leading to the upregulation of anti-angiogenic soluble vascular endothelial growth factor receptor 1 (sFlt-1). We have previously observed that Cajal bodies (CBs), subnuclear domains, are associated with the chromosome 19 miRNA gene cluster (C19MC), which encodes miR-517-3p. We have also found that coilin, the CB marker protein, is a positive regulator of miRNA biogenesis. Here we report that coilin is a regulator of miR-517-3p, sFlt-1, TNIP1, TNFSF15 and NF-κB activation, and this regulation is influenced by hypoxia. We also report that coilin and CBs are induced in the reduced uterine perfusion pressure (RUPP) rat model of PE. Collectively, the data presented here implicate coilin as a novel regulator of NF-κB activation and sFlt-1 upregulation.

Photoacoustic imaging provides an in vivo assessment of the preeclamptic placenta remodeling and function in response to therapy

IntroductionThere is a lack of effective therapeutic interventions for preeclampsia. A central factor in the etiology of the disease is the development of placental hypoxia due to abnormal vascular remodeling. However, methods to assess the impact of potential therapies on placental growth and remodeling are currently lacking. Here, we develop and validate ultrasound-guided photoacoustic imaging methods to monitor the placental response to therapeutic intervention. Establishing non-invasive tools to image placental function opens up previously unachievable understandings of placental therapeutic response. MethodsStudies were performed in the reduced uterine perfusion pressure (RUPP) rat model of preeclampsia. Preclinical research has identified tempol, a superoxide dismutase mimetic, and the phosphodiesterase inhibitor sildenafil as potential therapeutics for preeclampsia, as both improve in vivo maternal outcomes. PA images of the placental environment were acquired in RUPP rats receiving tempol (n = 8) or sildenafil (n = 8) to assess the longitudinal effects of treatment on placental oxygenation and vascular remodeling. Imaging measurements were validated with ex vivo histological analysis. ResultsSpectral photoacoustic imaging non-invasively measured placental hypoxia and impaired vascular growth two days after the RUPP procedure was implemented. Sildenafil significantly improved (p < 0.05) placental oxygenation and promoted vascular remodeling in RUPP animals, while RUPP animals treated with tempol had a diminished placental therapeutic response. DiscussionWe demonstrate that photoacoustic imaging provides in vivo measures of placental oxygenation and vascular remodeling, a previously unobtainable assessment of preeclamptic therapeutic response. These imaging tools have tremendous potential to accelerate the search for effective therapies for preeclampsia.

Nobiletin Inhibits Hypoxia-Induced Placental Damage via Modulating P53 Signaling Pathway.

In this study, we aimed to evaluate the effect of Nobiletin (NOB) on the placenta of Sprague–Dawley (SD) rats that had undergone reduced uterine perfusion pressure (RUPP) surgery and to evaluate the safety of NOB intervention during pregnancy. The results showed that NOB alleviated placental hypoxia, attenuated placental cell apoptosis, and inhibited placental damage in RUPP rats. No side effect of NOB intervention during pregnancy was observed. BeWo cell lines with P53 knockdown were then constructed using lentiviral transfection, and the P53 signaling pathway was found to be essential for NOB to reduce hypoxia-induced apoptosis of the BeWo cell lines. In summary, NOB attenuated hypoxia-induced placental damage by regulating the P53 signaling pathway, and those findings may contribute some insights into the role of NOB in placental development and the prevention of placental-related diseases.

Short-term effects of first trimester low-dose aspirin therapy on uterine artery flow in women at high risk for preeclampsia

The aim of the study was to investigate the short-term effects of low-dose aspirin treatment on uterine artery Doppler in pregnancies that were high-risk for preeclampsia. This prospective observational study included 82 patients with singleton pregnancies between 11–14 weeks of gestation. Uterine artery Doppler measurements were obtained by transvaginal ultrasonography at the first prenatal visit of women who started low-dose aspirin treatment due to the high risk of preeclampsia and women who did not receive low-dose aspirin. Uterine artery Doppler measurements of both groups were repeated 7–10 days after the first examination. There was a significant decrease in the presence of uterine artery diastolic notch in the low-dose aspirin group (p < .001). In both groups, the mean uterine artery PI values between the two exams tended to decrease, but the decrease in the control group was the only significant decrease (p = .014). The changes in Doppler indices were more pronounced in the control group. Therefore, they are much more likely to be related to the increase in gestational age than with the use of aspirin. Although there was an improvement in uterine arteries of Doppler measurements in high-risk women, they still had a high resistant flow compared to women with normal pregnancies. IMPACT STATEMENT What is already known on this subject? Pregnant women at high risk of preeclampsia have increased uterine artery resistance. Low-dose aspirin therapy is the only strategy to prevent the development of preeclampsia in these patients. What do the results of this study add? Low-dose aspirin therapy may improve uterine perfusion in the short term. After treatment, uterine artery resistance remains higher than in normal pregnancies. What are the implications of these findings for clinical practice and/or further research? Since baseline uterine blood flow and changes throughout pregnancy can be very different depending on the risk of preeclampsia, aspirin-treated or untreated groups may be used to evaluate the efficacy of aspirin in future studies. For example, patients with 11 weeks and 13 weeks of gestation can be divided into aspirin treated and control groups and efficacy of starting the aspirin treatment at 11 weeks of gestation vs 13 weeks of gestation can be evaluated.