Poorly controlled diabetes mellitus alters placental structure, efficiency, and plasticity

IntroductionThe hemochorial placenta provides a critical barrier at the maternal–fetal interface to modulate maternal immune tolerance and enable gas and nutrient exchange between mother and conceptus. Pregnancy outcomes are adversely affected by diabetes mellitus; however, the effects of poorly controlled diabetes on placental formation, and subsequently fetal development, are not fully understood.Research design and methodsStreptozotocin was used to induce hyperglycemia in pregnant rats for the purpose of investigating the impact of poorly controlled diabetes on placental formation and fetal development. The experimental paradigm of hypoxia exposure in the pregnant rat was also used to assess properties of placental plasticity. Euglycemic and hyperglycemic rats were exposed to ambient conditions (~21% oxygen) or hypoxia (10.5% oxygen) beginning on gestation day (gd) 6.5 and sacrificed on gd 13.5. To determine whether the interaction of hyperglycemia and hypoxia was directly altering trophoblast lineage development, rat trophoblast stem (TS) cells were cultured in high glucose (25 mM) and/or exposed to low oxygen (0.5% to 1.5%).ResultsDiabetes caused placentomegaly and placental malformation, decreasing placental efficiency and fetal size. Elevated glucose disrupted rat TS cell differentiation in vitro. Evidence of altered trophoblast differentiation was also observed in vivo, as hyperglycemia affected the junctional zone transcriptome and interfered with intrauterine trophoblast invasion and uterine spiral artery remodeling. When exposed to hypoxia, hyperglycemic rats showed decreased proliferation and ectoplacental cone development on gd 9.5 and complete pregnancy loss by gd 13.5. Furthermore, elevated glucose concentrations inhibited TS cell responses to hypoxia in vitro.ConclusionsOverall, these results indicate that alterations in placental development, efficiency, and plasticity could contribute to the suboptimal fetal outcomes in offspring from pregnancies complicated by poorly controlled diabetes.

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Characterization of the adverse effects of nicotine on placental development: in vivo and in vitro studies

AJP Endocrinology and Metabolism ◽

10.1152/ajpendo.00478.2013 ◽

2014 ◽

Vol 306 (4) ◽

pp. E443-E456 ◽

Cited By ~ 30

Author(s):

A. C. Holloway ◽

A. Salomon ◽

M. J. Soares ◽

V. Garnier ◽

S. Raha ◽

...

Keyword(s):

Adverse Effects ◽

Cell Line ◽

Angiogenic Factor ◽

Trophoblast Invasion ◽

Placental Development ◽

Nicotine Receptors ◽

Increased Risk ◽

The Impact

In utero exposure to nicotine is associated with increased risk of numerous adverse fetal and neonatal outcomes, which suggests that it acts directly to affect placental development and the establishment of the fetomaternal circulation (FC). This study used both in vivo [Wistar rats treated with 1 mg/kg nicotine from 2 wk prior to mating until gestational day (GD) 15] and in vitro (RCHO-1 cell line; treated with 10−9 to 10−3M nicotine) models to examine the effects of nicotine on these pathways. At GD 15, control and treated placentas were examined for the impact of nicotine on 1) trophoblast invasion, proliferation, and degree of hypoxia, 2) labyrinth vascularization, 3) expression of key genes of placental development, and 4) expression of placental angiogenic factors. The RCHO-1 cell line was used to determine the direct effects of nicotine on trophoblast differentiation. Our in vivo experiments show that nicotine inhibits trophoblast interstitial invasion, increases placental hypoxia, downregulates labyrinth vascularization as well as key transcription factors Hand1 and GCM1, and decreases local and circulating EG-VEGF, a key placental angiogenic factor. The in vitro experiments confirmed the inhibitory effects of nicotine on the trophoblast migration, invasion, and differentiation processes and demonstrated that those effects are most likely due to a dysregulation in the expression of nicotine receptors and a decrease in MMP9 activity. Taken together, these data suggest that adverse effects of maternal smoking on pregnancy outcome are due in part to direct and endocrine effects of nicotine on the main processes of placental development and establishment of FC.

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Maternal Oxidative Stress, Placental Morphometry, and Fetal Growth in Diabetic Rats Exposed to Cigarette Smoke

Reproductive Sciences ◽

10.1177/1933719118815589 ◽

2018 ◽

Vol 26 (9) ◽

pp. 1287-1293 ◽

Cited By ~ 4

Author(s):

Yuri K. Sinzato ◽

Estela M. Bevilacqua ◽

Gustavo T. Volpato ◽

Rogelio E. Hernandez-Pando ◽

Marilza V. C. Rudge ◽

...

Keyword(s):

Oxidative Stress ◽

Cigarette Smoke ◽

Fetal Growth ◽

Fetal Development ◽

Oxidative Stress Biomarkers ◽

Junctional Zone ◽

Pregnant Rats ◽

Stress Biomarkers ◽

Vascular Walls ◽

The Impact

The diabetic syndrome affects pregnancy, contributing to placental functional and structural disruptions and impaired fetal development, with many reports indicating tobacco-associated morbidity and perinatal mortality. In our study, an experimental rat model of diabetes and cigarette smoke exposure in pregnant rats was used to determine the impact of the combination of diabetes and exposure to cigarette smoke during pregnancy on maternal oxidative stress biomarkers and placental and fetal development. Diabetes was induced by streptozotocin, and dams were exposed to cigarette smoke by mainstream smoke generated by a mechanical smoking device and delivered into a chamber. Four groups of dams were studied: nondiabetic (C, control) and diabetic (D) exposed to filtered air and nondiabetic (CS) and diabetic (DS) exposed to cigarette smoke prior to and during pregnancy. Maternal oxidative stress biomarkers, placental morphology, and fetal growth were determined close to term. The combination of diabetes and cigarette smoke resulted in elevated maternal blood glucose levels and increased number of small fetuses. Placentas from the DS group showed increased junctional zone and decreased labyrinthine area. The morphological alterations were characterized by extensive vascular congestion, thickness, and hyalinization of the vascular walls, numerous decidual cells with abundant glycogen, and macrophages with cytoplasmic inclusions of hemosiderin. Additionally, they showed increased glycogen accumulation and junctional zone structural derangement with ectopic giant cells. No alterations were observed in maternal oxidative stress status. Thus, our result suggests that diabetes makes pregnant rats more susceptible to the adverse effects of exposure to cigarette smoke on placental morphometry and fetal growth.

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Embryotoxic impact of Zika virus in a rhesus macaque in vitro implantation model†

Biology of Reproduction ◽

10.1093/biolre/ioz236 ◽

2020 ◽

Vol 102 (4) ◽

pp. 806-816 ◽

Cited By ~ 4

Author(s):

Lindsey N Block ◽

Matthew T Aliota ◽

Thomas C Friedrich ◽

Michele L Schotzko ◽

Katherine D Mean ◽

...

Keyword(s):

Rhesus Macaque ◽

Pregnancy Outcomes ◽

Zika Virus ◽

Negative Impact ◽

Culture Media ◽

Blastocyst Formation ◽

Placental Development ◽

Zikv Infection ◽

The Impact

Abstract Zika virus (ZIKV) infection is associated with adverse pregnancy outcomes in humans, and infection in the first trimester can lead to miscarriage and stillbirth. Vertical and sexual transmissions of ZIKV have been demonstrated, yet the impact of infection during the initial stages of pregnancy remains unexplored. Here we defined the impact of ZIKV on early embryonic and placental development with a rhesus macaque model. During in vitro fertilization (IVF), macaque gametes were inoculated with a physiologically relevant dose of 5.48log10 plaque-forming units (PFU) of Zika virus/H.sapiens-tc/PUR/2015/PRVABC59_v3c2. Exposure at fertilization did not alter blastocyst formation rates compared to controls. To determine the impact of ZIKV exposure at implantation, hatched blastocysts were incubated with 3.26log10, 4.26log10, or 5.26log10 PFU, or not exposed to ZIKV, followed by extended embryo culture for 10 days. ZIKV exposure negatively impacted attachment, growth, and survival in comparison to controls, with exposure to 5.26log10 PFU ZIKV resulting in embryonic degeneration by day 2. Embryonic secretion of pregnancy hormones was lower in ZIKV-exposed embryos. Increasing levels of infectious virus were detected in the culture media post-exposure, suggesting that the trophectoderm is susceptible to productive ZIKV infection. These results demonstrate that ZIKV exposure severely impacts the zona-free blastocyst, whereas exposure at the time of fertilization does not hinder blastocyst formation. Overall, early stages of pregnancy may be profoundly sensitive to infection and pregnancy loss, and the negative impact of ZIKV infection on pregnancy outcomes may be underestimated.

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Molecular pathophysiology of diabetes mellitus during pregnancy with antenatal complications

Scientific Reports ◽

10.1038/s41598-020-76689-9 ◽

2020 ◽

Vol 10 (1) ◽

Author(s):

Arthur T. Kopylov ◽

Olga Papysheva ◽

Iveta Gribova ◽

Galina Kotaysch ◽

Lubov Kharitonova ◽

...

Keyword(s):

Diabetes Mellitus ◽

Lipid Metabolism ◽

Cord Blood ◽

Fetal Development ◽

Secondary Response ◽

Molecular Events ◽

Ppar Signaling ◽

Quantitative Alteration ◽

Molecular Pathophysiology ◽

The Impact

AbstractGestational diabetes mellitus is a daunting problem accompanied by severe fetal development complications and type 2 diabetes mellitus in postpartum. Diagnosis of diabetic conditions occurs only in the second trimester, while associated antenatal complications are typically revealed even later. We acquired an assay of peripheral and cord blood samples of patients with different types of diabetes mellitus who delivered either healthy newborns or associated with fetopathy complications. Obtained data were handled with qualitative and quantitative analysis. Pathways of molecular events involved in diabetes mellitus and fetopathy were reconstructed based on the discovered markers and their quantitative alteration. Plenty of pathways were integrated to differentiate the type of diabetes and to recognize the impact of the diabetic condition on fetal development. The impaired triglycerides transport, glucose uptake, and consequent insulin resistance are mostly affected by faulted lipid metabolism (APOM, APOD, APOH, APOC1) and encouraged by oxidative stress (CP, TF, ORM2) and inflammation (CFH, CFB, CLU) as a secondary response accompanied by changes in matrix architecture (AFM, FBLN1, AMBP). Alterations in proteomes of peripheral and cord blood were expectedly unequal. Both up- and downregulated markers were accommodated in the cast of molecular events interconnected with the lipid metabolism, RXR/PPAR-signaling pathway, and extracellular architecture modulation. The obtained results congregate numerous biological processes to molecular events that underline diabetes during gestation and uncover some critical aspects affecting fetal growth and development.

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MEF2 transcription factors in human placenta and involvement in cytotrophoblast invasion and differentiation

Physiological Genomics ◽

10.1152/physiolgenomics.00076.2017 ◽

2018 ◽

Vol 50 (1) ◽

pp. 10-19 ◽

Cited By ~ 5

Author(s):

Lucy Li ◽

Lewis P. Rubin ◽

Xiaoming Gong

Keyword(s):

Transcription Factors ◽

Human Placenta ◽

Cellular Proliferation ◽

Cell Types ◽

Dominant Negative ◽

Trophoblast Invasion ◽

Placental Development ◽

Cytotrophoblast Cell ◽

Adverse Pregnancy

Development of the human placenta and its trophoblast cell types is critical for a successful pregnancy. Defects in trophoblast invasion and differentiation are associated with adverse pregnancy outcomes, including preeclampsia. The members of myocyte enhancer factor-2 (MEF2) family of transcription factors are key regulators of cellular proliferation, differentiation, and invasion in various cell types and tissues and might play a similarly important role in regulating trophoblast proliferation, invasion, and differentiation during human placental development. In the present study, using human cytotrophoblast cell lines (HTR8/SVneo and BeWo) and primary human cytotrophoblasts (CTBs), we show that members of the MEF2 family are differentially expressed in human placental CTBs, with MEF2B and MEF2D being highly expressed in first trimester extravillous CTBs. Overexpression of MEF2D results in cytotrophoblast proliferation and enhances the invasion and migration of extravillous-like HTR8/SVneo cells. This invasive property is blocked by overexpression of a dominant negative MEF2 (dnMEF2). In contrast, MEF2A is the principal MEF2 isoform expressed in term CTBs, MEF2C and MEF2D being expressed more weakly, and MEF2B expression being undetected. Overexpression of MEF2A induces cytotrophoblast differentiation and syncytium formation in BeWo cells. During in vitro differentiation of primary CTBs, MEF2A expression is associated with CTB differentiation into syncytiotrophoblast. Additionally, the course of p38 MAPK and ERK5 activities parallels the increase in MEF2A expression. These findings suggest individual members of MEF2 family distinctively regulate cytotrophoblast proliferation, invasion, and differentiation. Dysregulation of expression of MEF2 family or of their upstream signaling pathways may be associated with placenta-related pregnancy disorders.

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Superoxide anion curbs nitric oxide modulation of afferent arteriolar ANG II responsiveness in diabetes mellitus

AJP Renal Physiology ◽

10.1152/ajprenal.2000.278.2.f302 ◽

2000 ◽

Vol 278 (2) ◽

pp. F302-F309 ◽

Cited By ~ 28

Author(s):

Gwynn C. Schoonmaker ◽

Rachel W. Fallet ◽

Pamela K. Carmines

Keyword(s):

Nitric Oxide ◽

Diabetes Mellitus ◽

Superoxide Anion ◽

Insulin Dependent Diabetes Mellitus ◽

Dependent Diabetes Mellitus ◽

Ang Ii ◽

Endogenous Nitric Oxide ◽

The Impact ◽

Arteriolar Diameter

Experiments were performed to test the hypothesis that the impact of endogenous nitric oxide (NO) on ANG II-induced renal arteriolar constriction is reduced in rats with insulin-dependent diabetes mellitus (65 mg/kg streptozotocin; STZ). Arteriolar diameter responses to exogenous ANG II were quantified before and during NO synthase inhibition (100 μM N ω-nitro-l-arginine;l-NNA) by using the in vitro blood-perfused juxtamedullary nephron technique. Afferent arteriolar lumen diameter averaged 20.7 ± 2.0 μm in Sham kidneys and 25.9 ± 1.3 μm in STZ kidneys ( P < 0.05). Efferent arteriolar diameter did not differ between Sham and STZ rats. In kidneys from Sham rats, afferent and efferent arteriolar responses to ANG II (0.1–10.0 nM) were exaggerated significantly by l-NNA. l-NNA also augmented efferent arteriolar ANG II responses in kidneys from STZ rats (high-glucose bath) but did not alter ANG II responses in afferent arterioles from STZ rats. l-NNA also accentuated efferent, but not afferent, arteriolar ANG II responses in STZ kidneys during acute restoration of bath glucose to normal levels. Superoxide dismutase (150 U/ml) restored the ability of l-NNA to allow exaggerated afferent arteriolar responses to ANG II in kidneys from STZ rats. These observations indicate that superoxide anion suppresses the modulatory influence of endogenous NO on ANG II-induced afferent arteriolar constriction in diabetes mellitus.

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Abstract P639: Sildenafil Treatment Improves Placental Levels of Placental Growth Factor in the Dahl Salt-Sensitive Pregnant Rat Model of Superimposed Preeclampsia

Hypertension ◽

10.1161/hyp.68.suppl_1.p639 ◽

2016 ◽

Vol 68 (suppl_1) ◽

Author(s):

Ana C Palei ◽

Jennifer M Sasser ◽

Joey P Granger

Keyword(s):

Growth Factor ◽

Umbilical Vein ◽

Resistance Index ◽

Placental Growth Factor ◽

Trophoblast Invasion ◽

Maternal Circulation ◽

Pregnant Rats ◽

Pregnant Rat

Although the etiology of preeclampsia (PE) remains unclear, evidence indicates that impaired trophoblast invasion followed by placental ischemia promotes the release of placental anti-angiogenic factors into the maternal circulation. These factors then elicit maternal endothelial dysfunction and hypertension by blocking the action of molecules such as the placental growth factor (PlGF). Inhibition of phosphodiesterase (PDE)-5 with sildenafil or other has been proposed as a potential therapy for PE; however, the mechanisms whereby PDE-5 inhibitors reduce blood pressure (BP) and improve uteroplacental perfusion during pregnancy are not clear. While previous studies have shown that PDE-5 inhibition induces PlGF production from human umbilical vein endothelial cells; it is unknown whether PDE-5 inhibitors also increase PlGF from placenta. Thus, the aim of this study was to evaluate whether sildenafil enhance placental secretion/production of PlGF in vitro and in vivo. In our in vitro protocol, we incubated placental villous explants from Sprague Dawley (SD) pregnant rats (n=4, 2-3 placentas per rat) at gestational day (GD)19 with different doses of sildenafil for 48h at 37°C under normoxia (8% O 2 ). PlGF-2 was measured in media of cultured explants by ELISA. We observed that sildenafil had no effect on PlGF-2 secretion from rat placental villi (vehicle: 562.7±46.6, 10nM: 559.3±39.5, 100nM: 556.4±35.9, 10uM: 546.2±37.5, and 100uM: 558.7±48.2pg/mg; P>0.05). In our in vivo protocol, we treated Dahl Salt-Sensitive (DS) pregnant rats (n=6-8 per group), which we had previously characterized as a model of superimposed PE, with sildenafil (50mg/kg per day, via food) from GD10 to 20. PlGF-2 was measured in placental homogenates by ELISA. While untreated DS dams exhibited an increase in BP and uterine artery resistance index (UARI) from baseline to late pregnancy, sildenafil-treated DS dams exhibited a significant decrease in BP and UARI. In addition, we found that placental levels of PlGF-2 were elevated in sildenafil-treated DS dams compared with untreated counterparts (1019±107.3 and 646.8±125.1pg/mg; P=0.0407). In conclusion, our findings suggest that the BP and UARI reduction in response to sildenafil may involve the indirect production of PlGF.

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Abstract 15820: Differential Myocardial Gene Expression of Glucose Transporters in Heart Transplant Recipients With and Without Diabetes Mellitus

Circulation ◽

10.1161/circ.142.suppl_3.15820 ◽

2020 ◽

Vol 142 (Suppl_3) ◽

Author(s):

Marc Vanderheyden ◽

Leen Delrue ◽

Sofie Verstreken ◽

Riet Dierckx ◽

Ward Heggermont ◽

...

Keyword(s):

Gene Expression ◽

Diabetes Mellitus ◽

Experimental Data ◽

Heart Transplant ◽

Glucose Transporters ◽

Myocardial Glucose Uptake ◽

Glucose Levels ◽

Elevated Glucose ◽

The Impact ◽

Hba1c Levels

Introduction: The Sodium Glucose cotransporter (SGLT) and glucose transporters(GLUTs) play a crucial role in cellular glucose transport. Although experimental data have shown differential regulation of GLUT4 and SGLT in diabetic cardiomyopathy, the impact of diabetes mellitus (DM) upon myocardial glucose transporters in humans remains undetermined. Aim: To better understand the impact of elevated glucose levels upon myocardial expression of glucose transporters, the endomyocardial gene expression of GLUT1, GLUT4 and SGLT1 was investigated in heart transplant(HTx) recipients, with and without DM, who received a heart from a DM- donor. Methods: At baseline(BL), immediately after HTx and 12 ± 2 months(FU) later, serial endomyocardial biopsies were procured in 26 Htx pts, free of clinical or histological rejection, at time of routine surveillance biopsy. Patients were categorized in DM+ (n = 13pts) and DM- (n = 13 pts), according to the presence of diabetes mellitus (DM) at FU. Results: Despite similar hemodynamics and HbA1c levels at BL, DM+ pts had higher HbA1c levels (46,00 ± 13,79 vs 38,33 ± 4,88; p < 0,05) at FU. No differences were noted in BL GLUT1, GLUT4 and SGLT gene expression between both groups. In DM- pts SGLT1(0,081 ± 0,080 vs 0,188 ± 0,108; p = 0,0036) , GLUT4(0,076 ± 0,068 vs 0,137 ± 0,065; p = 0,0011 )and GLUT1(0,020 ± 0,021 vs 0,022 ± 0,009; p = 0,043) increased significantly at FU whereas no change was observed in DM+ pts. Conclusion: Similar to experimental data, differential endomyocardial regulation in SGLT1 and GLUT4 was noted between DM+ and DM-pts with a blunted upregulation of glucose transporters at 1 year in DM+ HTx pts. These observations are in line with experimental data and suggest that myocardial glucose uptake is differentially regulated in DM+ HTx pts.

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Impact of Diabetes Mellitus on Bone Health

International Journal of Molecular Sciences ◽

10.3390/ijms20194873 ◽

2019 ◽

Vol 20 (19) ◽

pp. 4873 ◽

Cited By ~ 11

Author(s):

Murray ◽

Coleman

Keyword(s):

Diabetes Mellitus ◽

Fracture Risk ◽

Bone Health ◽

Assessment Tools ◽

Risk Assessment Tools ◽

Control Of Diabetes ◽

Structural Levels ◽

The Impact

Long-term exposure to a diabetic environment leads to changes in bone metabolism and impaired bone micro-architecture through a variety of mechanisms on molecular and structural levels. These changes predispose the bone to an increased fracture risk and impaired osseus healing. In a clinical practice, adequate control of diabetes mellitus is essential for preventing detrimental effects on bone health. Alternative fracture risk assessment tools may be needed to accurately determine fracture risk in patients living with diabetes mellitus. Currently, there is no conclusive model explaining the mechanism of action of diabetes mellitus on bone health, particularly in view of progenitor cells. In this review, the best available literature on the impact of diabetes mellitus on bone health in vitro and in vivo is summarised with an emphasis on future translational research opportunities in this field.

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Lipids from Oxidized Low-Density Lipoprotein Modulate Human Trophoblast Invasion: Involvement of Nuclear Liver X Receptors

Endocrinology ◽

10.1210/en.2003-1747 ◽

2004 ◽

Vol 145 (10) ◽

pp. 4583-4591 ◽

Cited By ~ 59

Author(s):

Laëtitia Pavan ◽

Axelle Hermouet ◽

Vassilis Tsatsaris ◽

Patrice Thérond ◽

Tatsuya Sawamura ◽

...

Keyword(s):

Nuclear Receptors ◽

Low Density Lipoprotein ◽

Density Lipoprotein ◽

Trophoblast Invasion ◽

Low Density ◽

Dependent Manner ◽

Placental Development ◽

Concentration Dependent Manner ◽

Human Trophoblast

Abstract Human embryonic implantation involves major invasion of the uterine wall and remodeling of the uterine arteries by extravillous cytotrophoblast cells (EVCT). Abnormalities in these early steps of placental development lead to poor placentation and fetal growth defects and are frequently associated with preeclampsia, a major complication of human pregnancy. We recently showed that oxidized low-density lipoproteins (oxLDLs) are present in situ in EVCT and inhibit cell invasion in a concentration-dependent manner. The aim of the present study was to better understand the mechanisms by which oxLDL modulate trophoblast invasion. We therefore investigated the presence of oxLDL receptors in our cell culture model of human invasive primary EVCT. We found using immunocytochemistry and immunoblotting that the lectin-like oxLDL receptor-1 was the scavenger receptor mainly expressed in EVCT and was probably involved in oxLDL uptake. We next examined the effect of low-density lipoprotein oxidative state on trophoblast invasion in vitro using EVCT cultured on Matrigel-coated Transwell. We demonstrated that only oxLDL containing a high proportion of oxysterols and phosphatidylcholine hydroperoxide derivatives that provide ligands for liver X receptor (LXR) and peroxisomal proliferator-activated receptor γ (PPARγ), respectively, reduced trophoblast invasion. We next investigated the presence and the role of these nuclear receptors and found that in addition to PPARγ, human invasive trophoblasts express LXRβ, and activation of these nuclear receptors by specific synthetic or natural ligands inhibited trophoblast invasion. Finally, using a PPARγ antagonist, we suggest that LXRβ, rather than PPARγ, is involved in oxLDL-mediated inhibition of human trophoblast invasion in vitro.

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