Platelets and endothelial dysfunction in gestational diabetes mellitus.

The prevalence of gestational diabetes mellitus (GDM) has increased in recent years, along with the higher prevalence of obesity in women of reproductive age. GDM is a pathology associated with vascular dysfunction in the fetoplacental unit. GDM-associated endothelial dysfunction alters the transfer of nutrients to the foetus affecting newborns and pregnant women. Various mechanisms for this vascular dysfunction have been proposed, of which the most studied are metabolic alterations of the vascular endothelium. However, different cell types are involved in GDM-associated endothelial dysfunction, including platelets. Platelets are small, enucleated cell fragments that actively take part in blood haemostasis and thrombus formation. Thus, they play crucial roles in pathologies coursing with endothelial dysfunction, such as atherosclerosis, cardiovascular diseases, and diabetes mellitus. Nevertheless, platelet function in GDM is understudied. Several reports show a potential relationship between platelet volume and mass with GDM; however, platelet roles and signaling mechanisms in GDM-associated endothelial dysfunction are unclear. This review summarizes the reported findings and proposes a link among altered amount, volume, mass, reactivity, and function of platelets and placenta development, resulting in fetoplacental vascular dysfunction in GDM.

The effect of Gestational Diabetes Mellitus on the fetal compartment.

In indicated preterm births such a Gestational Diabetes Mellitus (GDM), little is known about the role of the amnion membranes. Investigating the role of amnion membrane inflammation in response GDM may suggest novel pathophysiologic mechanisms. We hypothesize that increased GDM inflammatory mediators may weaken the amnion membrane predisposing them to infection. Maternal and fetal serum and amnion membrane biopsies were collected from 20 GDM and 38 normoglycemic subjects (control) who underwent elective cesarean sections. Cytokines and adipokines were evaluated in serum and amnion culture supernatant samples. Amnion membrane biopsies from GDM and control subjects were studied: fresh frozen for RNA analysis for Toll-like receptor expression; cultured with LPS to test membrane permeability, and inflammation LPS + anti-TLR4 for testing mechanism. GDM was associated with higher fetal serum leptin (p = 0.004) and IL-10 (p = 0.04) compared to controls. Amnion membrane explants from GDM had higher levels of IL-6 (p = 0.019), and lower expression of Claudin-4 (p = 0.007) and increased permeability (p = 0.046) compared to controls. GDM membranes treated with LPS showed an increased expression of IL-10 (p = 0.013); IL-6 (p = 0.004) and TNF-α (p = 0.0005) but did not affect membrane permeability. LPS and anti-TLR4 antibody treatment reduced the production of TNF-α in controls (p = 0.03) and GDM (p = 0.007) compared to LPS alone. Fetal inflammatory response seems more balanced in GDM and does not impact membrane permeability function even with an infectious stimulus. Light fetal membrane inflammatory response may explain lack of preterm labor in GDM. Concluding, benign inflammation in the membranes may not be harmful for pregnancy maintenance.

Stromal Cell-Derived Factor (SDF) 2 and the Endoplasmic Reticulum Stress Response of Trophoblast Cells in Gestational Diabetes Mellitus and In vitro Hyperglycaemic Condition.

BACKGROUND AND AIM: The endoplasmic reticulum (ER) stress response and the unfolded protein response (UPR) are essential cellular mechanisms to ensure the proper functioning of ER in adverse conditions. However, activation of these pathways has also been associated with insulin resistance and cell death in pathological conditions such as diabetes mellitus. In the present study, we investigated whether stromal cell-derived factor 2 (SDF2)-an ER stress-responsive factor-is related to ER response in placental cells exposed to maternal gestational diabetes mellitus (GDM) or to a hyperglycaemic in vitro condition. OBJECTIVE: The study aimed to investigate the role of SDF2 in BeWo cells , a trophoblast cell line originating from choriocarcinoma , and in placental tissue under hyperglycaemic conditions. METHODS: Protein levels of SDF2 and UPR factors, glucose-related protein 78 (GRP78) and eukaryotic initiation factor 2 alpha (elF2 alpha) were evaluated in the placentae of pregnant women diagnosed with GDM and treated by diet-control (insulin was added when necessary). The mRNA expression of SDF2 and UPR factors CHOP and sXBP1 were assessed in cultured BeWo cells challenged with glucose and treated with or without insulin. RESULTS: SDF2 expression was increased in the placentae of GDM women treated with diet. However, its values were similar to those of normoglycemic controls when the GDM women were treated with insulin and diet. BeWo cells cultured with high glucose and insulin showed decreased SDF2 expression, while high glucose increased CHOP and sXBP1 expression, which was then significantly reverted with insulin treatment. CONCLUSION: Our findings extend the understanding of ER stress and SDF2 expression in placentae exposed to hyperglycaemia, highlighting the relevance of insulin in reducing the levels of ER stress factors in placental cells. Understanding the effect of ER stress partners such as SDF2 on signalling pathways involved in gestation, complicated by hyperglycaemia, is pivotal for basic biomedical research and may lead to new therapeutic possibilities.

High levels of maternal total tri-iodothyronine, and low levels of fetal free L-thyroxine and total tri-iodothyronine, are associated with altered deiodinase expression and activity in placenta with gestational diabetes mellitus.

Gestational Diabetes Mellitus (GDM) is characterized by abnormal maternal D-glucose metabolism and altered insulin signaling. Dysregulation of thyroid hormones (TH) tri-iodethyronine (T3) and L-thyroxine (T4) Hormones had been associated with GDM, but the physiopathological meaning of these alterations is still unclear. Maternal TH cross the placenta through TH Transporters and their Deiodinases metabolize them to regulate fetal TH levels. Currently, the metabolism of TH in placentas with GDM is unknown, and there are no other studies that evaluate the fetal TH from pregnancies with GDM. Therefore, we evaluated the levels of maternal TH during pregnancy, and fetal TH at delivery, and the expression and activity of placental deiodinases from GDM pregnancies. Pregnant women were followed through pregnancy until delivery. We collected blood samples during 10-14, 24-28, and 36-40 weeks of gestation for measure Thyroid-stimulating hormone (TSH), Free T4 (FT4), Total T4 (TT4), and Total T3 (TT3) concentrations from Normal Glucose Tolerance (NGT) and GDM mothers. Moreover, we measure fetal TSH, FT4, TT4, and TT3 in total blood cord at the delivery. Also, we measured the placental expression of Deiodinases by RT-PCR, western-blotting, and immunohistochemistry. The activity of Deiodinases was estimated quantified rT3 and T3 using T4 as a substrate. Mothers with GDM showed higher levels of TT3 during all pregnancy, and an increased in TSH during second and third trimester, while lower concentrations of neonatal TT4, FT4, and TT3; and an increased TSH level in umbilical cord blood from GDM. Placentae from GDM mothers have a higher expression and activity of Deiodinase 3, but lower Deiodinase 2, than NGT mothers. In conclusion, GDM favors high levels of TT3 during all gestation in the mother, low levels in TT4, FT4 and TT3 at the delivery in neonates, and increases deiodinase 3, but reduce deiodinase 2 expression and activity in the placenta.

An extra virgin olive oil-enriched diet improves maternal, placental, and cord blood parameters in GDM pregnancies.

AIMS: To address the effect of a diet enriched in extra virgin olive oil (EVOO) on maternal metabolic parameters and placental proinflammatory markers in Gestational diabetes mellitus (GDM) patients. METHODS: Pregnant women at 24-28 weeks of gestation were enrolled: 33 GDM patients which were randomly assigned or not to the EVOO-enriched group and 17 healthy controls. Metabolic parameters were determined. Peroxisome proliferator activated receptor (PPAR) γ and PPARα protein expression, expression of microRNA (miR)-130a and miR-518d (which respectively target these PPAR isoforms) and levels of proinflammatory markers were evaluated in term placentas. Matrix metalloproteinases (MMPs) activity was evaluated in term placentas and umbilical cord blood. RESULTS: GDM patients that received the EVOO-enriched diet showed reduced pregnancy weight gain (GDM-EVOO:10.3 ± 0.9, GDM:14.2 ± 1.4, P = .03) and reduced triglyceridemia (GDM-EVOO:231 ± 14, GDM:292 ± 21, P = .02) compared to the non-EVOO-enriched GDM group. In GDM placentas, the EVOO-enriched diet did not regulate PPARγ protein expression or miR-130a expression, but prevented the reduced PPARα protein expression (P = .02 vs GDM) and the increased miR-518d expression (P = .009 vs GDM). Increased proinflammatory markers (interleukin-1ß, tumour necrosis factor-α and nitric oxide overproduction) in GDM placentas were prevented by the EVOO-enriched diet (respectively P = .001, P = .001 and P = .01 vs GDM). MMPs overactivity was prevented in placenta and umbilical cord blood in the EVOO-enriched GDM group (MMP-9: respectively P = .01 and P = .001 vs GDM). CONCLUSIONS: A diet enriched in EVOO in GDM patients reduced maternal triglyceridemia and weight gain and has antiinflammatory properties in placenta and umbilical cord blood, possibly mediated by the regulation of PPAR pathways.

Alterations in the structural characteristics of rectus abdominis muscles caused by diabetes and pregnancy: A comparative study of the rat model and women.

BACKGROUND AND OBJECTIVE: In the present study, we compared the effect of diabetic pregnancy on the rectus abdominis muscle (RAM) in humans and rats. We hypothesized that our animal model could provide valuable information about alterations in the RAM of women with Gestational Diabetes (GDM). METHOD: Newborns female rats (n = 10/group) were administered streptozotocin (100 mg/kg body weight) subcutaneously and were mated on reaching adulthood, to develop the mild hyperglycemic pregnant (MHP) rat model. At the end of pregnancy, the mothers were sacrificed, and the RAM tissue was collected. Pregnant women without GDM (non-GDM group; n = 10) and those diagnosed with GDM (GDM group; n = 8) and undergoing treatment were recruited, and RAM samples were obtained at C-section. The RAM architecture and the distribution of the fast and slow fibers and collagen were studied by immunohistochemistry. RESULTS: No statistically significant differences in the maternal and fetal characters were observed between the groups in both rats and women. However, significant changes in RAM architecture were observed. Diabetes in pregnancy increased the abundance of slow fibers and decreased fast fiber number and area in both rats and women. A decrease in collagen distribution was observed in GDM women; however, a similar change was not observed in the MHP rats. CONCLUSION: Our results indicated that pregnancy- associated diabetes- induced similar structural adaptations in the RAM of women and rats with slight alterations in fiber type number and area. These findings suggest that the MHP rat model can be used for studying the effects of pregnancy-associated diabetes on the fiber structure of RAM.

Placental exosomes viewed from an 'omics' perspective: implications for gestational diabetes biomarkers identification.

Exosomes are defined as extracellular vesicles that are released from cells upon fusion of an intermediate endocytic compartment - the multivesicular body - with the plasma membrane. Recently, placenta-derived exosomes have gained special attention, since they play a crucial role in the communication between the mother and fetus. It is known that the concentration of placenta-derived exosomes in the maternal bloodstream is higher in patients with preeclampsia or gestational diabetes mellitus. However, their composition in terms of the content of proteins, nucleic acids or lipids is uncertain. In this work, we reviewed the recent advances in placental exosomes characterization through omics-based methods, and their potential to predict gestational diabetes mellitus.

Late Cognitive Consequences of Gestational Diabetes to the Offspring, in a New Mouse Model.

Gestational diabetes mellitus (GD) is a form of insulin resistance triggered during gestation, which affects approximately 10% of pregnant women. Although previously considered a transient condition with few long-term consequences, growing evidence suggest that GD may be linked to permanent metabolic and neurologic changes in the offspring. Currently available GD models fail to recapitulate the full spectrum of this disease, thus providing limited information about the true burden of this condition. Here, we describe a new mouse model of GD, based on the administration of an insulin receptor antagonist (S961, 30 nmol/kg s.c. daily) during pregnancy. Pregnant mice developed increased fasting glycemia and glucose intolerance in the absence of maternal obesity, with a return to normoglycemia shortly after parturition. Moreover, we showed that the adult offspring of GD dams presented pronounced metabolic and cognitive dysfunction when exposed to short-term high-fat diet (HFD). Our data demonstrate that S961 administration to pregnant mice comprises a valuable approach to study the complex pathophysiology of GD, as well as strategies focused on prevention and treatment of both the mother and the offspring. Our findings suggest that the offspring of GD mothers are more susceptible to metabolic and cognitive impairments when exposed to high-fat diet later in life, thus indicating that approaches to prevent and treat these late effects should be pursued.

Morphological features of the human placenta and its free chorionic villi in normal pregnancies and those with diabetes and high blood pressure: literature review / Características morfológicas de la placenta humana y sus vellosidades coriales libres en gestas normales y con diabetes e hipertensión arterial: revisión de la literatura

The most prevalent pathologies during pregnancy, such as hypertension, gestational diabetes mellitus (GDM), and intrauterine growth restriction (IUGR), can determine modifications in macro- and microscopic morphological features of the placenta and its free chorionic villi. In the fetus it may be accompanied by pathological manifestations, with the embryo's future quality of life, and even its viability, at risk. The aim of this work is to perform a review of the morphological alterations that the placenta exhibits in pregnancies with GDM. A search on the topic in the scientific literature and specialized textbooks was carried out. The terms "placenta", "diabetes mellitus gestational" and "gestational diabetes mellitus" were used to define the search. Among the morphological modifications it was found that the placental weight/fetal weight ratio is higher in GDM and is associated mainly with histological changes. The distance between maternal and fetal circulation is increased because of an increase in the chorionic villi on the surface as well as greater thickness of the syncytiotrophoblast basal membrane due to an increased type IV collagen deposition. The stroma between the villi is edematous, which modifies the metabolic and endocrinal function of these placentas. Moreover, the capillary surface is enlarged due to the phenomena of vascular neoformation and a greater penetration of these vessels within the villi. Low oxygen partial pressure (pO2) was detected, which would produce a compensatory hyperplasia of terminal chorionic villi. In GDM the placenta undergoes alterations in its formation, structure, and function. According to the review, these alterations are related to an oxygenation deficiency in the fetus and changes in the transplacental transport of nutrients and other alterations, causing fetal overgrowth by increasing their availability in addition to other consequences to the developing fetus. In the case of high blood pressure during pregnancy, this produces accelerated maturation and rapid aging of the chorionic villi, with the risk of inducing a placental abruption. In addition, placental circulation is reduced by a third, decreasing oxygen saturation in the umbilical vessels and placing the health of the fetus at risk.

Las patologías de la gestación más prevalentes, como hipertensión, diabetes mellitus gestacional (DMG) y restricción del crecimiento uterino, pueden determinar modificaciones en las características morfológicas macro y microscópicas de la placenta y sus vellosidades coriales libres,y en el feto se puede acompañar de manifestaciones patológicas, con riesgo para su calidad de vida futura, e incluso su viabilidad. El objetivo de este trabajo consiste en realizar una revisión acerca de las alteraciones morfológicas que presenta la placenta en gestaciones con DMG. Se llevó a cabo una búsqueda sobre la temática en literatura científica y libros de texto especializados. Se utilizaron los términos "placenta", "diabetes mellitus gestacional" y "gestational diabetes mellitus" para orientar la búsqueda. Entre las modificaciones morfológicas se encontró que la relación peso placentario/peso fetal está aumentado en la DMG y se asocia principalmente a cambios histológicos. La distancia entre la circulación materna y la fetal está aumentada debido a un aumento de la superficie entre las vellosidades coriales, así como al mayor grosor de la membrana basal del sincitiotrofoblasto por un depósito elevado de colágeno tipo IV. El estroma entre las vellosidades se encuentra edematoso lo que va a modificar la función metabólica y endocrina de estas placentas. También hay aumento de la superficie capilar por fenómenos de neoformación vascular y una mayor penetración de estos vasos dentro de las vellosidades. Se detectó baja presión parcial de oxígeno (pO2), que produciría una hiperplasia compensatoria de vellosidades coriónicas terminales. En la DMG la placenta experimenta alteraciones en su formación, estructura y función. Según lo revisado, estas alteraciones están relacionadas con déficit en la oxigenación del feto, cambios en el transporte transplacentario de los nutrientes y otras alteraciones, ocasionando sobrecrecimiento fetal por aumento de su disponibilidad, entre otras consecuencias al feto en desarrollo. Y en el caso de la hipertensión arterial durante la gestación produce una maduración acelerada y envejecimiento rápido de las vellosidades coriales, con el riesgo de inducir un desprendimiento prematuro de placenta. Además se reduce la circulación placentaria en un tercio, disminuyendo la saturación de oxígeno en los vasos umbilicales del, poniendo en riesgo la salud del feto.

Aspectos morfocuantitativos de las vellosidades coriales libres en gestas normales, con diabetes, hipertensión arterial y restricción del crecimiento intrauterino / Morphoquantitative characteristics of free corial vellosities in normal births, with diabetes, hypertension and restriction of intrauterine growth

Las patologías de la gestación como la hipertensión, diabetes mellitus gestacional, o restricción del crecimiento intrauterino, pueden determinar modificaciones en las características morfológicas macro y microscópicas de la placenta y sus vellosidades coriales libres, y en el feto se puede acompañar de manifestaciones patológicas, con riesgo para su calidad de vida futura, e incluso su viabilidad. El objetivo de este trabajo consistió en describir aspectos morfométricos e histológicos de las vellosidades coriales libres en gestas normales, con diabetes e hipertensión arterial. Se utilizaron 30 placentas humanas y fueron separadas, según presencia o ausencia de patologías en el embarazo, en tres grupos: Normal (N), Síndrome Hipertensivo del Embarazo (SHE), Diabetes (D) y Restricción del Crecimiento Intrauterino (RCIU). Se usó ficha para registrar peso placentario y del recién nacido Todas las muestras fueron fijadas en formalina tamponada al 10 %. De cada una fueron extraídas 5 muestras, obteniendo 25 cortes por cada placenta. Posteriormente, fueron teñidas con H&E, Azul Alcián y Tricrómico de Masson. Además, se efectuó el análisis histológico y morfométrico (ImageJ®) de las vellosidades coriales. El análisis estadístico fue realizado utilizando ANOVA. Entre los cambios morfológicos, se encontró una relación peso placentario/peso del recién nacido aumentada en la Diabetes Mellitus Gestacional asociada a cambios histológicos. No hubo cambios morfométricos significativos entre placentas N, SHE y D. Hubo un aumento en el número de vasos coriales en placentas del grupo D (P < 0,05) y de la superficie entre las vellosidades coriales. En el grupo SHE hubo aumento moderado de nudos sinciciales y presencia de fibrina en el estroma. Las placentas con Diabetes Mellitus Gestacional experimentan alteraciones histológicas, como consecuencia de cambios estructurales y funcionales. Además, el aumento de vasos sanguíneos en placentas con diabetes se produce por neoformación vascular y mayor penetración de vasos sanguíneos dentro de las vellosidades. En el caso del SHE las alteraciones placentarias se relacionan con la gravedad de la enfermedad.

Gestational pathologies such as hypertension, gestational diabetes mellitus and restriction of intrauterine growth can determine changes in the macro and microscopic morphological characteristics of the placenta and its free chorionic villi. In the fetus it can be accompanied by pathological manifestations with risk to its viability and future quality of life. The aim of this work was to describe morphometric and histological aspects of free chorionic villi in normal pregnancies associated with diabetes, hypertension and restriction of intrauterine growth. Thirty human placentas were used and were separated into three groups: Normal (N), Hypertensive Pregnancy Syndrome (SHE), Diabetes (D), and Restriction of Intrauterine Growth (RIG) according to evident pathologies or absence thereof during pregnancy. Tab was used to record placental and newborn weight. All samples were fixed in 10 % buffered formalin. From each, 5 samples were extracted, obtaining 25 cuts for each placenta. Subsequently, they were stained with H & E, Alcian Blue and Masson's Trichrome. In addition, histological and morphometric analysis (ImageJ®) of the chorion villus was carried out. Statistical analysis was performed using ANOVA. Among the morphological changes, an increased placental weight / weight ratio of the newborn was found in Gestational Diabetes Mellitus associated with histological changes. There were no significant morphometric changes between placentas N, SHE and D. There was an increase in the number of corial vessels in placentas of group D (P <0.05) and of the surface between the chorion villi. In the SHE group there was a moderate increase in syncytial nodes and presence of fibrin in the stroma. Placentas with Gestational Diabetes Mellitus experience histological alterations, as a consequence of structural and functional changes. In addition, the increase of blood vessels in placentas D is produced by vascular neoformation and increased penetration of blood vessels into the villi. In the case of SHE, placental alterations are related to the severity of the disease.

Histomorphological study of placenta in gestational diabetes mellitus / Estudio histomorfológico de la placenta en diabetes mellitus gestacional

Evidence from the literature shows that well-controlled glucose levels during pregnancy are usually associated with normal placental morphology. The aim of this study was to identify the lacental changes attributed to maternal hyperglycemia. A total of 20 placentae were selected for study from a tertiary care medical center in Makkah city, Saudi Arabia. Out of 20, 10 placentae were from patients diagnosed with GDM based on IADSPG criteria, and 10 placentae were from patients with normal pregnancies without GDM. The morphometric measurements were recorded. The mean weight of GDM placentae were more than the normal placentae. Upon histopathology, significant changes such as syncytial knots, cytotrophoblastic cell proliferation, fibrinoid necrosis, stromal fibrosis, and hyalinized villi were observed in GDM placentae. GDM produces significant morphological alterations in the placentae, which might affect the developing fetus.

La evidencia de la literatura muestra que niveles de glucosa bien controlados durante el embarazo generalmente se asocian con una morfología placentaria normal. El objetivo de este estudio fue identificar los cambios placentarios atribuidos a la hiperglucemia materna. Un total de 20 placentas fueron seleccionadas para un estudio en un centro médico de atención terciaria en la ciudad de La Meca, Arabia Saudita. De 20 placentas, 10 de estas fueron de pacientes diagnosticadas con diabetes mellitus gestacional (DMG) según los criterios de IADSPG, y 10 placentas fueron de pacientes con embarazos normales sin DMG. Las mediciones morfométricas fueron registradas. El peso medio de las placentas GDM fue mayor que la placenta normal. Tras la histopatología, se observaron cambios significativos tales como nudos sincitiales, proliferación celular citotrofoblástica, necrosis fibrinoide, fibrosis estromal y vellosidades hialinizadas en placenta con DMG. La DMG produce alteraciones morfológicas significativas en las placentas, que pueden afectar al desarrollo del feto.

Circular RNA expression profiles in placental villi from women with gestational diabetes mellitus.

Circular RNAs (circRNAs) have recently been shown to exert their effects on multiple pathological processes by acting as microRNA (miRNA) sponges. However, the roles of circRNAs in gestational diabetes mellitus (GDM) are largely unknown. This study aimed to identify the circRNAs involved in GDM and predict their potential biological functions. We first performed next-generation sequencing (NGS) to generate unbiased placental villi circRNA expression profiles of GDM and normal controls. In total, 48,270 circRNAs from the placental villi of the two groups were sequenced. Of these, 227 circRNAs were significantly up-regulated and 255 circRNAs were significantly down-regulated. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) biological pathway analyses demonstrated that glycometabolism and lipometabolism processes, which are important in GDM development, were significantly enriched. Further analysis showed that most of the circRNAs harbored miRNA binding sites, and some were associated with GDM. These results showed that circRNAs are aberrantly expressed in the placental villi of GDM patients and play potential roles in the development of GDM.

Maternal insulin therapy does not restore foetoplacental endothelial dysfunction in gestational diabetes mellitus.

Pregnant women diagnosed with gestational diabetes mellitus subjected to diet (GDMd) that do not reach normal glycaemia are passed to insulin therapy (GDMi). GDMd associates with increased human cationic amino acid transporter 1 (hCAT-1)-mediated transport of L-arginine and nitric oxide synthase (NOS) activity in foetoplacental vasculature, a phenomenon reversed by exogenous insulin. Whether insulin therapy results in reversal of the GDMd effect on the foetoplacental vasculature is unknown. We assayed whether insulin therapy normalizes GDMd-associated foetoplacental endothelial dysfunction. Primary cultures of human umbilical vein endothelial cells (HUVECs) from GDMi pregnancies were used to assay L-arginine transport kinetics, NOS activity, p44/42mapk and protein kinase B/Akt activation, and umbilical vein rings reactivity. HUVECs from GDMi or GDMd show increased hCAT-1 expression and maximal transport capacity, NOS activity, and eNOS, and p44/42mapk, but not Akt activator phosphorylation. Dilation in response to insulin or calcitonin-gene related peptide was impaired in umbilical vein rings from GDMi and GDMd pregnancies. Incubation of HUVECs in vitro with insulin (1 nmol/L) restored hCAT-1 and eNOS expression and activity, and eNOS and p44/42mapk activator phosphorylation. Thus, maternal insulin therapy does not seem to reverse GDMd-associated alterations in human foetoplacental vasculature.

Gestational diabetes induces pancreatic beta-bells apoptosis in adult rat offspring / Diabetes gestacional induce apoptosis de células beta del páncreas en crías de ratas adultas

Several studies indicated that pancreatic ß-cell death occurs in both type 1 and type 2 diabetes. This experimental study was designed to determine the effect of gestational diabetes on the ß-cells in 16-week-old rat offspring. By this aim, adult Wistar rats aged 10-12 weeks were randomly allocated in control and diabetic groups. The diabetic group received 40 mg/kg/body weight of streptozotocin (STZ) on day zero of gestation. After delivery, diabetic offspring of GDM mothers and controls at the age of 16 weeks were sacrificed and pancreases harvested and fixed. The number of ß-cells and were counted by Gomori's method staining. Also, apoptosis in pancreas tissue of diabetic and control offspring was detected by TUNEL assay. Results showed a significant reduction in ß-cell number in offspring of GDM (p<0.05). TUNEL assay showed that the number of apoptotic cells increased in GDM compared to controls (P<0.05). This study revealed that gestational diabetes induces pancreatic beta-cells apoptosis in 16-week-old rat offspring.

Varios estudios indican que la muerte de las células ß del páncreas se produce tanto en la diabetes Tipo 1 como en la Tipo 2. Este estudio experimental fue diseñado para determinar el efecto de la diabetes gestacional en las células ß del páncreas en crías de ratas de 16 semanas. Para ello, ratas Wistar adultas de entre 10-12 semanas fueron asignadas al azar en dos grupos: control y diabetes. El grupo diabetes recibió 40 mg / kg / peso corporal de estreptozotocina (STZ) en el día cero de la gestación. Después del parto, a las 16 semanas, las crías de las madres diabéticas y controles de madres con diabetes gestacional (MDG), fueron sacrificadas para la extracción del páncreas, el cual posteriormente fue fijado. Se contó el número de células ß del páncreas mediante tinción con el método de Gomori. Además, se detectó apoptosis en el tejido del páncreas de la descendencia diabética y el grupo control mediante un ensayo TUNEL. Los resultados mostraron una reducción significativa en el número de células b en la descendencia de MDG (p <0,05). El ensayo TUNEL mostró que el número de células apoptóticas aumentó en MDG en comparación con los controles (P <0,05). Este estudio reveló que la diabetes gestacional induce apoptosis de células ß en el páncreas de crías de ratas de 16 semanas.

The effect of gestational diabetes mellitus on sciatic nerve in adult offspring rats / Efecto de la diabetes mellitus gestacional sobre el nervio ciático en ratas adultas

Gestational diabetes mellitus (GDM) is one form of diabetes affect approximately 7 % of pregnancies. Diabetic peripheral neuropathy (DPN) is a common complication of diabetes that is associated with loss of nerve fibers, myelin abnormalities and significant decrease in the expression of myelin basic protein (MBP) in peripheral nerves. This study was done to determine the effect of induced diabetes during pregnancy on sciatic nerve in adult rat offspring. In this study, wistar rats' dams were allocated to control and diabetic groups. Diabetic rats were received 40 mg/kg/body weight of streptozotocin (STZ) on the first day of gestation. Six offspring of each group were randomly selected on 12 weeks postnatal and histopathological changes in their nerve tissue were examined through H&E staining and transmission electron microscopy. Furthermore, the expression of MBP in sciatic nerve was examined by immunohistochemistry. We found that the myelinated fiber number of sciatic nerve in offspring of diabetic rats was reduced compared to the controls, but this difference was not significant. The average thickness of the myelin sheath of sciatic nerve fibers in the control and GDM was 97.1±0.1and 94.1±0.2 µm, respectively that the difference was not statistically significant. The expression of MBP protein in the myelin sheath of both groups was similar. TEM results showed that myelin sheath of diabetic offspring had not any changes compared to control. Atrophy of axons and schwannocytus (Schwann cells) alterations were not observed in diabetic offspring. Induction of diabetes during pregnancy reduced the number of nerve fibers and thickness of the myelin sheath. But it has no effect on MBP expression and schwannocytus morphology.

La diabetes mellitus gestacional (DMG) es una forma de diabetes que afecta aproximadamente al 7 % de los embarazos. La neuropatía periférica diabética (NPD) es una complicación frecuente de la diabetes asociada a la pérdida de fibras nerviosas, anomalías de la mielina y disminución significativa de la expresión de la proteína básica de mielina (PBM) en los nervios periféricos. Este estudio se realizó para determinar el efecto de la diabetes inducida durante el embarazo en el nervio ciático en descendientes de ratas adultas. Las ratas Wistar madres fueron asignadas a los grupos control y diabéticas. Las ratas diabéticas recibieron 40 mg/kg/peso corporal de estreptozotocina (STZ) el primer día de gestación. Seis descendientes de cada grupo fueron seleccionados al azar en la semana 12 postnatal y los cambios histopatológicos en su tejido nervioso se examinaron a través de tinción H-E y microscopía electrónica de transmisión. Además, la expresión de PBM en el nervio ciático se examinó mediante inmunohistoquímica. Se encontró que el número de fibras mielinizadas de nervio ciático en descendientes de ratas diabéticas se redujo en comparación con los controles, pero esta diferencia no fue significativa. El espesor medio de la vaina de mielina de las fibras nerviosas ciáticas en el control y DMG fue de 97,1±0,1 y 94,1±0,2 µm, respectivamente, y la diferencia no fue estadísticamente significativa. La expresión de la proteína PBM en la vaina de mielina de ambos grupos fue similar. Los resultados del TEM mostraron que la vaina de mielina de la descendencia diabética no tuvo ningún cambio en comparación con el control. La atrofia de los axones y las alteraciones de los schwannocitos (células de Schwann) no se observaron en descendientes diabéticos. La inducción de diabetes durante el embarazo redujo el número de fibras nerviosas y el grosor de la vaina de mielina. Pero no tiene ningún efecto sobre la expresión de PBM y la morfología de las schwannocitos.

Histopathological and immunohistochemical changes in the amniotic membrane of gestational diabetic mothers / Cambios histopatológicos e inmunohistoquímicos de la membrana amniótica de madres diabéticas gestacionales

The aim of this study is to examine the changes in the amniotic membrane diagnosed with gestational diabetes mellitus. In this study, as a control group human amnion membrane from normotensive pregnancies was collected from diabetic women at 28­35 weeks of gestation. Gestational diabetes (n= 6) and normal amnion membrane (n= 6) for a total of 12 units were received. Amniotic membrane thickness was measured (p<0.0001) and it was significantly higher in GDM groups compared to control group. The diameter of the amniotic epithelial cell nuclei was measured (p=0.0022). Gestational diabetes results show that there was weakening between amniotic epithelial cell-cell junction. This study showed that structural changes in epithelial cells of amniotic membrane were formed due to diabetes. The membrane thickness has led to structural changes in diameter and in diabetes group cause extracellular matrix to increase, thus leading to MMP-9 expression increase eventually disrupting matrix balance. In addition, with cd44 increase angiogenesis has been induced and thought to influence material pass between fetus and mother.

El objetivo de este estudio fue examinar los cambios en la membrana amniótica diagnosticada con diabetes mellitus gestacional (DMG). En este estudio, como grupo control, se recogió la membrana amniótica de embarazos normotensos de mujeres diabéticas a las 28 y 35 semanas de gestación. La muestra consistió en 6 casos con diabetes gestacional (n = 6) y 6 casos de membrana amniótica normal (n = 6), para un total de 12 casos. El espesor de la membrana amniótica se midió (p <0,0001) y fue significativamente mayor en los grupos de DMG en comparación con el grupo control. Se midió el diámetro de los núcleos de las células epiteliales amnióticas (p = 0,0022). Los resultados demostraron que en la DMG hubo debilitamiento entre la célula epitelial amniótica-célula de unión. Este estudio mostró que los cambios estructurales en las células epiteliales de la membrana amniótica se presentaron debido a la DMG. El espesor de la membrana ha dado lugar a cambios estructurales en el diámetro y en el grupo de DMG debido a un aumento de la matriz extracelular, lo que condujo al aumento de la expresión de MMP-9, eventualmente interrumpiendo el equilibrio de la matriz. Además, el aumento de cd44 indujo la angiogénesis y se cree que también influye en el material que se comparte entre el feto y la madre.

Streptozotocin-Induced Maternal Hyperglycemia Increases the Expression of Antioxidant Enzymes and Mast Cell Number in Offspring Rat Ventral Prostate.

Gestational diabetes mellitus (GDM) has increased in recent years. Although the cellular and molecular mechanisms involved in GDM-increased risk factors to offspring remained poorly understood, some studies suggested an association between an increase in oxidative stress induced by maternal hyperglycemia and complications for both mothers and newborns. Here, we investigated the impact of maternal hyperglycemia followed by maternal insulin replacement during lactation on the expression of antioxidant enzymes and mast cell number in offspring ventral prostate (VP) at puberty. Pregnant rats were divided into three groups: control (CT); streptozotocin-induced maternal hyperglycemia (MH); and MH plus maternal insulin replacement during lactation (MHI). Male offspring were euthanized at postnatal day (PND) 60 and the VP was removed and processed for histology and Western blotting analyses. Maternal hyperglycemia delayed prostate maturation, and increased mast cell number catalase (CAT), superoxide dismutase (SOD), glutatione-s-transferase (GST-pi), and cyclooxygenase-2 (Cox-2) expression in the offspring of hyperglycemic dams. Maternal insulin replacement restored VP structure, mast cell number and antioxidant protein expression, except for Cox-2, which remained higher in the MHI group. Thus, an increase in oxidative stress induced by intrauterine hyperglycemia impacts prostate development and maturation, which persists until puberty. The overall improvement of maternal metabolism after insulin administration contributes to the restoration of prostate antioxidant enzymes and secretory function. Taken together, our results highlighted that imbalanced physiological maternal-fetal interaction contributes to the impairment of reproductive performance of the offspring from diabetic mothers. Anat Rec, 300:291-299, 2017. © 2016 Wiley Periodicals, Inc.

Influence of the Expression of Inflammatory Markers on Kidney after Fetal Programming in an Experimental Model of Renal Failure.

Objective. To evaluate the expression of inflammatory markers in experimental renal failure after fetal programming. Methods. The offspring aged two and five months were divided into four groups: CC (control dams, control offspring); DC (diabetic dams, control offspring); CFA (control dams, folic acid offspring, 250 mg/Kg); and DFA (diabetic dams, folic acid offspring). Gene expression of inflammatory markers MCP-1, IL-1, NOS3, TGF-ß, TNF-α, and VEGF was evaluated by RT-PCR. Results. MCP-1 was increased in the CFA and DFA groups at two and five months of age, as well as in DC5 when compared to CC5. There was a higher expression of IL-1 in the CFA2, DFA2, and DC2 groups. There was a decrease in NOS3 and an increase in TNF-α in DFA5 in relation to CFA5. The gene expression of TGF-ß increased in cases that had received folic acid at two and five months, and VEGF decreased in the CFA5 and DFA5 groups. DC5 showed increased VEGF expression in comparison with CC5. Conclusions. Gestational diabetes mellitus and folic acid both change the expression of inflammatory markers, thus demonstrating that the exposure to harmful agents in adulthood has a more severe impact in cases which underwent fetal reprogramming.

Gestational diabetes reduces pancreatic beta cells in rat offspring / La diabetes gestacional reduce las células beta pancreáticas en crías de ratas

Previous study has shown the adverse effects of gestational diabetes on hippocampal and spinal cord neuronal density in animal model. This study was conducted to determine the effect of gestational diabetes on beta cells in rat pancreas in early postnatal life. In this experimental study, 10 dams randomly allocated into control and diabetic groups on day 1 of gestation. Five dams in diabetic group received 40 mg/kg/BW of streptozotocin (intraperitoneally) and control animals received normal saline. Six of 28 and 56-day-old offspring of each gestational diabetes mellitus and controls were randomly scarified and sections were taken from the pancreas and stained using Gomorra's method. The density of beta cells and number and area of pancreatic islets were evaluated by quantitative computer-assisted morphometric method. The density of beta cells of 28-day-old offspring pancreas significantly reduced from 96.23±5.0 in control group to 71.5±5.3 cells in 10000 mm2 area of islet in diabetic group (P <0.01). The number of the pancreatic islets of in gestational diabetes (15.25±3.7) significantly reduced in comparison with the controls (8.61±0.7). The density of beta cells of 56-day-old offspring pancreas significantly reduced from 105.33±8.6 in control group to 62.12±5.9 in diabetic group (P <0.01). The number of the pancreatic islets of in gestational diabetes (13.5±0.5) significantly reduced compared to controls (6.75±1.7) (P <0.01). This study revealed that gestational diabetes loss the number of the beta cells in 28 and 56-day-old rat offspring.

Estudios previos han mostrado los efectos adversos de la diabetes gestacional en la densidad neuronal del hipocampo y de la médula espinal en modelos animales. Este estudio se llevó a cabo para determinar el efecto de la diabetes gestacional en las células beta del páncreas de rata en vida postnatal temprana. En este estudio experimental, 10 ratas fueron asignadas al azar a los grupos control y diabético en el día 1 de gestación. Cinco ratas del grupo diabético recibieron 40 mg/kg/BW de estreptozotocina (intraperitonealmente), mientras que los animales del grupo control recibieron solución salina normal. Seis de los descendientes, de 28 y 56 días de edad, de cada grupo, diabetes mellitus gestacional y control, se escarificaron al azar y se tomaron secciones del páncreas, que se tiñeron usando el método de Gomorra. La densidad de las células beta y el número y área de islotes pancreáticos fueron evaluados a través de método cuantitativo asistido por computadora morfométrica. La densidad de células beta del páncreas en las crías de 28 días disminuyó significativamente de 96,23 ± 5,0 en el grupo de control a 71,5 ± 5,3 células en el grupo diabético, en 10000 mm2 de área de islote (P <0,01). El número de islotes pancreáticos de la diabetes gestacional (15,25 ± 3,7) se redujo significativamente en comparación con los controles (8,61 ± 0,7). La densidad de células beta del páncreas en las crías de 56 días de edad se redujo de 105,33 ± 8,6 en el grupo de control a 62,12 ± 5,9 en el grupo diabético (P <0,01). El número de islotes pancreáticos en el grupo de diabetes gestacional (13,5 ± 0,5) se redujo significativamente en comparación con los controles (6,75 ± 1,7) (P <0,01). Este estudio reveló que la diabetes gestacional provoca una pérdida en el número de células beta en crías de ratas de 28 y 56 días de edad.

Validation of a symphysis-fundal height chart developed for pregnancy complicated by diabetes and hyperglycemia: an observational study.

BACKGROUND: The present study validates a symphysis-fundal height chart (SFH-chart) for pregnant women with type 2 diabetes mellitus (DM2), gestational diabetes mellitus (GDM) and mild gestational hyperglycemia (MGH) attending at the Diabetes and Pregnancy Reference Service of the Botucatu Medical School, UNESP, Brazil. METHODS: A cross-sectional study was carried out to evaluate the performance of the specific FHC in predicting small (SGA) and large (LGA) for gestational age newborns (NB). We evaluated 206 pregnant women with DM2, GDM or MGH and their NB. The last symphysis-fundal height measure, taken at birth, was used to determine the sensitivity index (Sens), specificity index (Spe), positive prediction value (PPV), negative prediction value (NPV) and accuracy in predicting SGA and LGA. The gold standard was the Lubchenco birth weight/gestational age ratio evaluated at birth. RESULTS: The mothers showed adequate glycemic control; 91.3 % of all pregnant women achieved HbA1c < 6,5 % in the third trimester. The SFH-chart tested achieved 100 % of Sens and NPV in predicting both SGA and LGA, with accuracy of 90.3 % (85.5; 93.6) and 91.8 % (87.2; 94.8), respectively, for predicting SGA and LGA newborns. CONCLUSIONS: The Basso SFH-chart showed high performance in predicting both SGA and LGA newborns of DM-2, GDM and MGH mothers, with better performance than the national reference SFH-chart. These findings support the internal validation of the Basso SFH-chart, which may be implemented in the prenatal care of the Diabetes and Pregnancy Reference Service-Botucatu Medical School/UNESP.