Gestational Diabetes Mellitus Placentas Research Articles

From Mother to Child: Epigenetic Signatures of Hyperglycemia and Obesity during Pregnancy.

In utero exposure to maternal hyperglycemia and obesity can trigger detrimental effects in the newborn through epigenetic programming. We aimed to assess the DNA methylation levels in the promoters of MC4R and LPL genes from maternal blood, placenta, and buccal swab samples collected in children born to mothers with and without obesity and Gestational Diabetes Mellitus (GDM). A total of 101 Caucasian mother-infant pairs were included in this study. Sociodemographic characteristics, clinical parameters, physical activity, and adherence to the Mediterranean diet were evaluated in the third trimester of pregnancy. Clinical parameters of the newborns were recorded at birth. A negative relationship between MC4R DNA methylation on the fetal side of the GDM placenta and birth weight (r = -0.630, p = 0.011) of newborns was found. MC4R DNA methylation level was lower in newborns of GDM women (CpG1: 2.8% ± 3.0%, CpG2: 3.8% ± 3.3%) as compared to those of mothers without GDM (CpG1: 6.9% ± 6.2%, CpG2: 6.8% ± 5.6%; p < 0.001 and p = 0.0033, respectively), and it was negatively correlated with weight (r = -0.229; p = 0.035), head circumference (r = -0.236; p = 0.030), and length (r = -0.240; p = 0.027) at birth. LPL DNA methylation was higher on the fetal side of the placenta in obese patients as compared to normal-weight patients (66.0% ± 14.4% vs. 55.7% ± 15.2%, p = 0.037), and it was associated with maternal total cholesterol (r = 0.770, p = 0.015) and LDL-c (r = 0.783, p = 0.012). These results support the role of maternal MC4R and LPL methylation in fetal programming and in the future metabolic health of children.

MiR-142-5p mediated Nrf2 dysregulation in gestational diabetes mellitus and its impact on placental angiogenesis

Gestational diabetes mellitus (GDM) presents significant risks during pregnancy, including adverse perinatal outcomes and placental dysfunction. Impaired angiogenesis, involving crucial factors like Vascular Endothelial Growth Factor (VEGF), contributes to these complications. The Nrf2/Keap1 pathway, crucial for vascular redox homeostasis, has been linked to GDM-associated angiogenesis dysregulation. This study aimed to investigate the molecular mechanisms underlying placental Nrf2 regulation, focusing on angiomiRs, key regulators of angiogenesis in GDM. Computational analysis identified miR-142-5p targeting Nrf2 mRNA. Expression levels of miR-142-5p were assessed in GDM placenta and correlated with Nrf2 expression. Experimental validation utilized human trophoblastic cell lines (BeWo) exposed to hyperglycemic conditions, assessing the effects of anti-miR-142 transfection on Nrf2 expression and angiogenic marker levels. miR-142-5p expression was significantly downregulated in GDM placenta, correlating positively with Nrf2 expression. In BeWo cells exposed to hyperglycemia, anti-miR-142 transfection notably increased Nrf2 expression alongside angiogenic marker levels, confirming the computational predictions. Our findings highlight the pivotal role of miRNAs in GDM-associated impaired angiogenesis by modulating Nrf2 expression. Understanding these molecular mechanisms provides insights into potential therapeutic targets for improving pregnancy outcomes in GDM cases.

Lysophosphatidylcholine Impairs the Mitochondria Homeostasis Leading to Trophoblast Dysfunction in Gestational Diabetes Mellitus.

Gestational diabetes mellitus (GDM) is a common pregnancy disorder associated with an increased risk of pre-eclampsia and macrosomia. Recent research has shown that the buildup of excess lipids within the placental trophoblast impairs mitochondrial function. However, the exact lipids that impact the placental trophoblast and the underlying mechanism remain unclear. GDM cases and healthy controls were recruited at Kaohsiung Medical University Hospital. The placenta and cord blood were taken during birth. Confocal and electron microscopy were utilized to examine the morphology of the placenta and mitochondria. We determined the lipid composition using liquid chromatography-mass spectrometry in data-independent analysis mode (LC/MSE). In vitro studies were carried out on choriocarcinoma cells (JEG3) to investigate the mechanism of trophoblast mitochondrial dysfunction. Results showed that the GDM placenta was distinguished by increased syncytial knots, chorangiosis, lectin-like oxidized low-density lipoprotein (LDL) receptor-1 (LOX-1) overexpression, and mitochondrial dysfunction. Lysophosphatidylcholine (LPC) 16:0 was significantly elevated in the cord blood LDL of GDM patients. In vitro, we demonstrated that LPC dose-dependently disrupts mitochondrial function by increasing reactive oxygen species (ROS) levels and HIF-1α signaling. In conclusion, highly elevated LPC in cord blood plays a pivotal role in GDM, contributing to trophoblast impairment and pregnancy complications.

AKT1 and MAPK8: New Targets for Gestational Diabetes Mellitus?

Objective: Gestational diabetes mellitus (GDM) disrupts placental function and increases risks for pregnancy. This study investigates the potential involvement of AKT1 and MAPK8 genes, known for their roles in insulin resistance and cell signaling, in GDM pathophysiology. Methods: Placental tissues from GDM patients and healthy controls were analyzed using real-time PCR to quantify gene expression levels. In silico analysis further explored the functional implications of expression changes. Results: AKT1 and MAPK8 displayed significantly altered expression in GDM placentas compared to controls (p = 0.047 and p = 0.007, respectively). In silico analysis suggests potential functional consequences related to diabetes-associated pathways. Conclusion: This study identifies differential expression of AKT1 and MAPK8 in GDM placentas, suggesting their potential roles in the disease process. Further investigation into their functional contributions could provide valuable insights into GDM pathophysiology and potential therapeutic targets.

Effect of insulin versus dietary restriction alone on umbilical cord and placental arterio-venous anastomoses in women with gestational diabetes mellitus

Background: Gestational diabetes mellitus (GDM) induces structural alterations in the umbilical cord and placental vessels. This study aims to understand the potential impact of insulin therapy on arteriovenous anastomosis (AVA) in placenta and umbilical cord. Materials and methods: We collected 52 normal and 59 GDM placentas and umbilical cords from the labour room. GDM specimens were categorized into those treated with diet (GDM-Diet) (n = 23) and those treated with insulin (GDM-Insulin) (n= 36). The umbilical cord was dissected near its insertion to expose the vessels, and arteriovenous anastomoses were documented. Tissues underwent processing and staining for microscopic confirmation of AV anastomosis. Comparison was done between normal and GDM specimens, and notably, between GDM-Diet and GDM-Insulin specimens. Results: Our examination of 59 GDM and 52 normal placentas with attached umbilical cords revealed AV anastomoses in 24 cases. AV anastomosis was observed in 22 (37.3%, n= 59) GDM cases and 2 (3.8%, n=52) normal cases (p=0.0001). Among GDM-Insulin group umbilical cords, 10 (27.78%, n = 36) exhibited AV anastomoses, while GDM-diet specimens displayed a notably higher proportion, with 12 (52.17%, n = 23) demonstrating AV anastomoses (p = 0.05). Conclusion: AV anastomoses are markedly more prevalent in the umbilical cords and placentas of individuals with GDM. Interestingly, within the GDM cohort, prevalence is significantly higher in those managed with diet. Our study highlights the association between GDM management strategies and AVC prevalence, emphasizing the importance of considering vascular dynamics in the management of GDM pregnancies. Keywords: placenta; umbilical cord; fetal- maternal health; gestational diabetes mellitus

Serotonin receptor and serotonin transporter expressions in the placental villous tree in gestational diabetes mellitus

BACKGROUND: The serotonergic system during pregnancy plays an important role not only in carbohydrate metabolism, but also in the laying and regulation of the fetoplacental complex, growth and development of the fetus. The study of the expression of placental serotonin 5-HT2A receptor and serotonin transporter (SERT) in gestational diabetes mellitus is foremost for scrutinizing the pathogenesis of perinatal complications, as it may allow for finding new opportunities for their prevention and correction.&#x0D; AIM: The aim of this study was to compare the expression patterns of the serotonin 5-HT2A receptor and SERT in placental tissue in gestational diabetes mellitus and in normal pregnancy.&#x0D; MATERIALS AND METHODS: This comparative cohort study included pregnant women with gestational diabetes mellitus (n = 6) and patients with normal pregnancy (n = 10). The expression of serotonin 5-HT2A receptor (Abcam, USA) and SERT (Bioss Antibodies, USA) was studied in placenta samples from the both study groups by immunohistochemical method. Morphometric analysis was performed using the VideoTest-Morphology 5.2 program (Videotest Ltd., Russia).&#x0D; RESULTS: The relative area of SERT expression in the placenta in gestational diabetes mellitus was higher compared to normal pregnancy (p 0.001). The relative areas of expression of the serotonin 5-HT2A receptor in the placenta did not differ between the study groups (p = 0.5).&#x0D; CONCLUSIONS: Higher SERT expression in the placentas of patients with gestational diabetes mellitus compared to those from women with normal pregnancies may reflect the level of tension of compensatory mechanisms in gestational diabetes mellitus and the effect of insulin therapy on these mechanisms.

Sex-Specific Alterations in Placental Proteomics Induced by Intrauterine Hyperglycemia.

Gestational diabetes mellitus (GDM) with intrauterine hyperglycemia induces a series of changes in the placenta, which have adverse effects on both the mother and the fetus. The aim of this study was to investigate the changes in the placenta in GDM and its gender differences. In this study, we established an intrauterine hyperglycemia model using ICR mice. We collected placental specimens from mice before birth for histological observation, along with tandem mass tag (TMT)-labeled proteomic analysis, which was stratified by sex. When the analysis was not segregated by sex, the GDM group showed 208 upregulated and 225 downregulated proteins in the placenta, primarily within the extracellular matrix and mitochondria. Altered biological processes included cholesterol metabolism and oxidative stress responses. After stratification by sex, the male subgroup showed a heightened tendency for immune-related pathway alterations, whereas the female subgroup manifested changes in branched-chain amino acid metabolism. Our study suggests that the observed sex differences in placental protein expression may explain the differential impact of GDM on offspring.

Protein Profiling of Placental Extracellular Vesicles in Gestational Diabetes Mellitus.

Throughout pregnancy, some degree of insulin resistance is necessary to divert glucose towards the developing foetus. In gestational diabetes mellitus (GDM), insulin resistance is exacerbated in combination with insulin deficiency, causing new-onset maternal hyperglycaemia. The rapid reversal of insulin resistance following delivery strongly implicates the placenta in GDM pathogenesis. In this case-control study, we investigated the proteomic cargo of human syncytiotrophoblast-derived extracellular vesicles (STBEVs), which facilitate maternal-fetal signalling during pregnancy, in a UK-based cohort comprising patients with a gestational age of 38-40 weeks. Medium/large (m/l) and small (s) STBEVs were isolated from GDM (n = 4) and normal (n = 5) placentae using ex vivo dual-lobe perfusion and subjected to mass spectrometry. Bioinformatics were used to identify differentially carried proteins and mechanistic pathways. In m/lSTBEVs, 56 proteins were differently expressed while in sSTBEVs, no proteins reached statistical difference. Differences were also observed in the proteomic cargo between m/lSTBEVs and sSTBEVs, indicating that the two subtypes of STBEVs may have divergent modes of action and downstream effects. In silico functional enrichment analysis of differentially expressed proteins in m/lSTBEVs from GDM and normal pregnancy found positive regulation of cytoskeleton organisation as the most significantly enriched biological process. This work presents the first comparison of two populations of STBEVs' protein cargos (m/l and sSTBEVs) from GDM and normal pregnancy isolated using placenta perfusion. Further investigation of differentially expressed proteins may contribute to an understanding of GDM pathogenesis and the development of novel diagnostic and therapeutic tools.

ABHD5-CPT1B: An Important Way of Regulating Placental Lipid Metabolism in Gestational Diabetes Mellitus

Gestational diabetes mellitus (GDM) is one of the most common metabolic disorders in pregnancy, and a novel association of maternal lipid profile has been suggested to play an important role. However, the molecular mechanism is not clear. Bio-analyzed combined with placental metabonomics and single-cell RNA-sequencing (scRNA-seq) successfully identified a potentially important molecule: α-β hydrolase domain-containing protein 5 (ABHD5). The syncytiotrophoblast (SCT) cell model was adopted as a fusion of BeWo cells in response to forskolin. On this basis, the high glucose-stimulated cell experiment was carried out. 15 women with GDM and 15 normal pregnant women were recruited for validation experiments. ABHD5 was mainly expressed in the trophoblast cells, especially in SCT cells, and significantly decreased in the GDM placenta. After stimulation by high glucose, the expression of ABHD5 was downregulated in a time-dependent manner in BeWo cells treated with forskolin. At the same time, lipid droplets (LDs) were increased in the SCT. LD storage was also increased in the SCT with siABHD5, while it was significantly reduced in SCT cells with high ABHD5 expression. However, this effect could be attenuated by downregulated carnitine palmitoyltransferase 1B (CPT1B). ABHD5-CPT1B is confirmed as an important regulator of placental lipid metabolism.

Galectin-10 Expression in Placentas of Women with Gestational Diabetes.

Galectins are known to play an important role in immunoregulatory processes and autoimmune diseases. Galectin-10 is a cytoplasmic protein of human eosinophils and is involved in various eosinophilic diseases. Since increased galectin expression is already detected in the placentas of mothers with gestational diabetes mellitus (GDM), this study focuses on the specific role of galectin-10 and hints at consequences for the diagnosis and therapeutic options of GDM. It is hypothesized that the difference in galectin-10 expression will raise the pathophysiological understanding of gestational diabetes. The study population consists of 80 women: 40 healthy mothers and 40 women suffering from gestational diabetes mellitus. The expression of galectin-10 was analyzed in the syncytiotrophoblast (SCT) and the decidua of the placenta via immunohistochemistry and immunofluorescence double staining. The immunoreactivity score (IRS) was used for evaluation. The results in this study were significant for an overexpression of galectin-10 in GDM placentas compared with the control group. The syncytiotrophoblast showed overexpression in the nucleus and the cytoplasm, whereas expression of galectin-10 in the decidua was significant in the cytoplasm only. This study identified the expression changes in galectin-10 in placental tissue between healthy and GDM mothers and intensified the understanding of gestational diabetes. Assuming that gestational diabetes mellitus is involved in inflammatory processes, galectin-10 might play a role in the development and maintenance of GDM. Further investigation is required to strengthen these findings.

MORPHOFUNCTIONAL AND MOLECULAR CHANGES IN PLACENTA AND PERIPHERAL BLOOD IN PREECLAMPSIA AND GESTATIONAL DIABETES MELITUS

Gestational diabetes mellitus (GDM) and preeclampsia (PE) are common pregnancy complications with similar risk factors. Although GDM is associated with PE, the exact mechanism underlying the association of these diseases is unclear. The aim of the work: to study morphofunctional and molecular changes in the placenta and peripheral blood both in PE and GDM. In the study, we evaluated local and systemic changes in the production of several placental proteins, markers of inflammation and metabolic disorders. Changes in villi expression of placental lactogen, trophoblastic β1-glycoprotein, placental alpha-1-microglobulin, and proteinase3 were noted in the groups of complicated pregnancies. The present study shows the similarity of the pathogenic mechanisms underlying PE and GDM.

An in vitro Model to Study Placental Functions in Gestational Diabetes Mellitus

Background: Gestational diabetes mellitus (GDM) is a serious pregnancy complication that affects around 14 % of pregnancies worldwide. It not only causes short-term complications in the mother and child but also significantly increases the risks of several chronic diseases, including cardiovascular diseases and diabetes. An excess production of the soluble vascular endothelial growth factor (VEGF) receptor 1 in the placenta, which is a natural inhibitor of VEGF, has been linked to various pregnancy complications. However, placentas affected by GDM often exhibit increased vascularization, suggesting that the role of VEGF in GDM differs from its impact on typical pregnancy complications. Unfortunately, limited studies on placental functions in GDM have been conducted due to a lack of reliable culture models. Methods: In this study, we developed an in vitro model of GDM using the human choriocarcinoma trophoblast cell line BeWo and examined the effects of VEGF on glucose uptake. The BeWo cells were treated with a high glucose-containing (25 mM) syncytialization differentiation medium to produce insulin-resistant cells. Insulin resistance was assessed by a glucose uptake assay using the fluorescent glucose analog 2-NBDG and flow cytometry analysis. Results: The cells treated with high glucose exhibited a significant decrease in glucose uptake, suggesting the successful development of glucose resistance in these cells. However, VEGF treatment did not show a significant impact on glucose uptake in high glucose-treated cells. Conclusions: It is important to note that glucose homeostasis is a complex process involving multiple cell types, and further studies are necessary to fully understand the functions of VEGF on GDM placentas. Nevertheless, the in vitro model we have developed and validated will be a valuable tool for studying placental pathophysiology in GDM and evaluating the effectiveness of potential therapeutic agents. HIGHLIGHTS BeWo cells are the choriocarcinoma cell line which were cultured in the in vitro condition BeWo cells were differentiated using forskolin and 8-Br Camp Insulin resistant model of BeWo cells were prepared by high glucose (25 mM) treatment The effect of VEGF on glucose homeostasis in GDM model of BeWo cells was observed GRAPHICAL ABSTRACT

Regulatory mechanisms underlying endoplasmic reticulum stress involvement in the development of gestational diabetes mellitus entail the CHOP-PPARα–NF–κB pathway

ObjectiveWe investigated the proinflammatory functions of endoplasmic reticulum stress and peroxisome proliferator-activated receptor α (PPARα) in the development of gestational diabetes mellitus (GDM) and their relationship in regulating inflammation in GDM. MethodsThis study was performed on placentas of normal pregnant women, women with GDM, and HTR8 cells. Transmission electron microscopy, immunohistochemistry, Western blot analysis, and RT-PCR were performed to analyze ERS and PPARα expression on both normal and GDM pregnancy placentas. ELISA was performed to analyze inflammatory biomarkers. To generate models of the GDM-like state, placentas of normal pregnancy were treated with LPS and polyinosinic-polycytidylic acid (poly [I:C]). TG, CHOP plasmid, and CHOP siRNA were assessed as to their regulation of HTR8 cells to discern the relationship between ERS and PPARα in regulating the inflammation associated with GDM. ResultsERS was elevated in GDM placentas, induced the secretion of IL-6 and TNF-α, and attenuated the expression of GLUT-4. PPARα was diminished in GDM placentas and inhibited the inflammatory responses via the NF-κB nuclear-transport process. 4-PBA reduced CHOP and augmented PPARα, and it decreased IL-6 and TNF-α in our GDM-like explant. However, with both 4-PBA and MK886 treatment, we noted no significant difference in CHOP expression. The level of PPARα was reduced, and that of NF-κB p65 in the nucleus was elevated with TG treatment in the HTR8/Svneo. Knockdown of CHOP increased PPARα and reduced NF-κB p65, while expression of PPARα declined, and that of NF-κB p65 rose with the application of CHOP when HTR8 cells were treated with TG. ConclusionsERS contributes to the pathophysiology of GDM in pregnancy via the CHOP-PPARα–NF–κB-signalling pathway by inducing aberrant activation of inflammation and insulin resistance.

Regulation of uterus and placenta remodeling under high estradiol levels in gestational diabetes mellitus models†.

The present study aimed to investigate the regulation of placentas and uterus remodeling and involvement of E2 in gestational diabetes mellitus (GDM). To achieve this, we established in vitro and in vivo models for GDM placentas by culturing BeWo cells under hyperglycemic concentration, and treating pregnant rats with streptozotocin (STZ). We evaluated the expression of angiogenesis-related proteins. The expression of the anti-angiogenic factor, excess placental soluble fms-like tyrosine kinase 1 (sFLT1) was increased in our in vitro GDM model compared to the control. Moreover, the expressions of placental sFLT1 and the von willebrand factor (VWF) were also significantly elevated in the placenta of STZ-treated rats. These data indicate the disruption of angiogenesis in the GDM placentas. The expression levels of connexin 43 (Cx43), a component of the gap junction and collagen type I alpha 2 chain (Col1a2), a component of the extracellular matrix, were decreased in the GDM uterus. These results suggest that uterus decidualization and placental angiogenesis are inhibited in GDM rats. Our results also showed upregulation of the expression of genes regulating E2 synthesis as well as estrogen receptors (ER) in vivo models. Accordingly, the concentration of E2 measured in the culture medium under hyperglycemic conditions, as well as in the serum and placenta of the STZ-treated rats, was significantly elevated compared to the control groups. These results suggest that the dysregulated remodeling of the placenta and uterus may result in the elevation of E2 and its signaling pathway in the GDM animal model to maintain pregnancy.

Maternal elevated inflammation impairs placental fatty acids β-oxidation in women with gestational diabetes mellitus.

An adverse proinflammatory milieu contributes to abnormal cellular energy metabolism response. Gestational diabetes mellitus (GDM) is closely related to an altered maternal inflammatory status. However, its role on lipid metabolism regulation in human placenta has not yet been assessed. The aim of this study was to examine the impact of maternal circulating inflammatory mediators ([TNF]-α, [IL]-6, and Leptin) on placental fatty acid metabolism in GDM pregnancies. Fasting maternal blood and placental tissues were collected at term deliveries from 37 pregnant women (17 control and 20 GDM). Molecular approach techniques as radiolabeled lipid tracers, ELISAs, immunohistochemistry and multianalyte immunoassay quantitative analysis, were used to quantify serum inflammatory factors' levels, to measure lipid metabolic parameters in placental villous samples (mitochondrial fatty acid oxidation [FAO] rate and lipid content [Triglycerides]), and to analyze their possible relationships. The effect of potential candidate cytokines on fatty acid metabolism in ex vivo placental explants culture following C-section a term was also examined. Maternal serum IL-6, TNF-α and leptin levels were significantly increased in GDM patients compared with control pregnant women (9,9±4,5 vs. 3,00±1,7; 4,5±2,8 vs. 2,1±1,3; and 10026,7±5628,8 vs. 5360,2±2499,9 pg/ml, respectively). Placental FAO capacity was significantly diminished (~30%; p<0.01), whereas triglyceride levels were three-fold higher (p<0.01) in full-term GDM placentas. Uniquely the maternal IL-6 levels showed an inverse and positive correlation with the ability to oxidize fatty acids and triglyceride amount in placenta, respectively (r= -0,602, p=0.005; r= 0,707, p=0.001). Additionally, an inverse correlation between placental FAO and triglycerides was also found (r=-0.683; p=0.001). Interestingly, we ex vivo demonstrated by using placental explant cultures that a prolonged exposure with IL-6 (10 ng/mL) resulted in a decline in the fatty acid oxidation rate (~25%; p=0.001), along to acute increase (2-fold times) in triglycerides accumulation (p=0.001), and in lipid neutral and lipid droplets deposits. Enhanced maternal proinflammatory cytokines levels (essentially IL-6) is closely associated with an altered placental fatty acid metabolism in pregnancies with GDM, which may interfere with adequate delivery of maternal fat across the placenta to the fetus.

Comparative study of microvascular structural changes in the gestational diabetic placenta.

Microvascular morphology and pathological changes in gestational diabetes mellitus (GDM) placentas and normal placentas were observed via vascular casting technology, electron microscopy, and pathological detection technology. Vascular structure and histological morphology changes in GDM placentas were examined to generate basic experimental data for the diagnosis and prognostic determination of GDM. This case-control study involving 60 placentas, 30 from healthy controls and 30 from patients with GDM. Differences in size, weight, volume, umbilical cord diameter, and gestational age were assessed. Histological changes in the placentas in the two groups were analyzed and compared. A placental vessel casting model was constructed using a self-setting dental powder technique, to compare the two groups. The placental cast microvessels of the two groups were compared using scanning electron microscopy. There were no significant differences in maternal age or gestational age between the GDM group and the control group (p > .05). The size, weight, volume, and thickness of the placentas in the GDM group were significantly greater than those in the control group, as was umbilical cord diameter (p < .05). Immature villus, fibrinoid necrosis, calcification, and vascular thrombosis were significantly greater in the placental mass in the GDM group (p < .05). The terminal branches of the microvessels in diabetic placenta casts were sparse, with significantly fewer ends and lower villous volume (p < .05). Gestational diabetes can cause gross and histological changes in the placenta, particularly placental microvascular changes.

High TXNIP expression accelerates the migration and invasion of the GDM placenta trophoblast

IntroductionOur previous study has proofed the glucose sensitive gene-thioredoxin-interacting protein (TXNIP) expression was up in the placenta of the patients with gestational diabetes mellitus (GDM), but the pathological mechanisms underlying abnormal TXNIP expression in the placenta of patients with GDM is completely unclear and additional investigations are required to explain the findings we have observed. In the present study, we simulated the high TXNIP expression via introducing the Tet-On “switch” in vitro, approximate to its expression level in the real world, to explore the following consequence of the abnormal TXNIP.MethodsThe expression and localization of TXNIP in the placenta of GDM patients and the health control was investigated via immunofluorescent staining, western blot and RT-qPCR. Overexpression of TXNIP was achieved through transfecting Tet-on system to the human trophoblastic cell line-HTR-8/Svneo cell. TXNIP knockout was obtained via CRISPR-Cas9 method. The cell phenotype was observed via IncuCyte Imaging System and flow cytometry. The mechanism was explored via western blot and RT-qPCR.ResultsThe expression level of TXNIP in the GDM placenta was nearly 2–3 times higher than that in the control. The TXNIP located at trophoblastic cells of the placenta. When the expression of TXNIP was upregulated, the migration and invasion of the cells accelerated, but cell apoptosis and proliferation did not changed compared with the control group. Furthermore, the size of the TetTXNIP cells became larger, and the expression level of Vimentin and p-STAT3 increased in the TetTXNIP cells. All the changes mentioned above were opposite in the TXNIP-KO cells.ConclusionsAbnormal expression of TXNIP might be related to the impairment of the GDM placental function, affecting the migration and invasion of the placental trophoblast cells through STAT3 and Vimentin related pathway; thus, TXNIP might be the potential therapeutic target for repairing the placental dysfunction deficient in GDM patients.

Histological and Morphological Study of the Placenta in Gestational: Pregestational Diabetic, and Normal Women’s

This study was aimed at comparing the morphometric and histological features of the placenta in gestational diabetes mellitus (GDM), pre-gestational diabetes mellitus (PGDM), and non-diabetic normal pregnant women. Twenty-four placentas were obtained from non-diabetic pregnant women (control group), ten from women with GDM, and ten from PGDM women. Segments from the central part of the placenta were taken and processed for paraffin blocks, sectioned, and stained with H&amp;E, Masson trichrome, and periodic acid-Schiff. The results of morphometric measurement revealed that the mean weight and diameter of GDM and PGDM placentae were significantly higher than in the control; placental thickness was a significant increase in PGDM; the number of cotyledons and fetal weights showed no significant difference in all these groups; the fetoplacental ratio was significantly higher in the PGDM; discoidal placenta shape, followed by irregular, oval, and kidney as the most common shapes recorded in these groups; the central umbilical cord insertion was most common in all three groups; followed by noncentral (eccentric) and marginal insertions. Diabetes caused several histopathological changes in the placenta, including an increase in the number of different sizes of villi with a decrease in the intervillous spaces, an increase in the: number of big syncytial knots, and fibrinoid necrosis; chronogenesis in the intervillous spaces and villous capillary congestion. Diabetic groups showed the presence of dense collagen fibers within the villous stroma (villous stromal fibrosis) and around fetal blood capillaries. GDM showed a mild thickening of the basement membrane (BM) of chorionic villi and a mild increase in the collagen in the villous stroma. While PGDM showed a moderate increase in the BM of syncytiotrophoblast and villous stroma and mild thickening of the BM of capillary blood vessels. In conclusion, both GDM and PGDM caused adverse effects on the histological and morphometrical features of the placenta. Keywords: Placenta; Diabetes Mellitus; Morphometric analysis; Histopathology, Masson trichrome stain; Periodic acid shift reaction. DOI: 10.7176/JBAH/13-6-03 Publication date: April 30 th 2023

The role of chemerin in the regulation of cGAS-STING pathway in gestational diabetes mellitus placenta.

Recent studies already confirmed that placenta mitochondrial dysfunction is associated with the progression of gestational diabetes mellitus (GDM). Besides, a possible relationship between adipokine chemerin and disulfide-bond A oxidoreductase-like protein (DsbA-L) had been revealed, whereas the potential interaction remains unclear. In addition, very little is still known about the cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) signaling pathway and its mechanisms of action in the context of GDM. The present study aims to investigate the underlying mechanism of cGAS-STING pathway and its regulatory relationship with chemerin in GDM. A total of 50 participants, including 25 cases of GDM patients and 25 pregnant women with normal glucose tolerance, were enrolled, and their placenta tissues at term labor were collected. Besides, an insulin resistance cell model was established on the human trophoblastic cell line to explore the molecular mechanism of chemerin on cGAS-STING pathway. Results showed that there were mitochondrial pathological changes in GDM placenta, accompanied by the decreased expression of DsbA-L, increased level of chemerin, and the activation of cGAS-STING pathway. In the insulin resistant cell model, overexpression of chemerin upregulated protein expression of DsbA-L, and recombinant chemerin presented time-dependent inhibition on the cGAS-STING pathway, but this effect was not dependent on DsbA-L. In conclusion, elevated chemerin is probably a protective mechanism, which may be a potential therapeutic strategy for GDM.

Gestational Diabetes Mellitus and Small-for-Gestational-Age: An Insight into the Placental Molecular Biomarkers.

Gestational diabetes mellitus (GDM) and small-for-gestational-age (SGA) are two metabolic-related diseases that could affect women during pregnancy. Considering that the chorionic villi (CVs) are crucial structures for the feto-maternal exchange, the alterations in their conformation have been linked to an imbalanced metabolic environment of placenta. In this study, a multidisciplinary approach has been carried out to describe the changes occurring in the placental CVs of GDM and SGA patients. The results revealed higher levels of superoxide dismutase 1 (SOD-1) and catalase (CAT), especially in the GDM placentae, which could be correlated with the hyperglycemic environment characteristic of this pathology. Furthermore, spectroscopy and histologic analyses revealed that both pathologies modify the placental lipid composition altering its structure. However, SGA induces lipid peroxidation and reduces collagen deposition within the CVs. Since the endocannabinoid system (ECS) is involved in placentation and different metabolic activities, the cannabinoid receptor 1 (CB1) and transient receptor potential cation channel subfamily V member 1 (TRPV-1) were analyzed. No changes have been observed either at general or specific levels in the CVs comparing control and pathological samples, suggesting the non-involvement of the cannabinoid system in these two pathologies.