Intrauterine Growth Retardation Rats Research Articles

Azithromycin exposure during pregnancy disturbs the fetal development and its characteristic of multi-organ toxicity

AimsAzithromycin is widely used in clinical practice for treating maternal infections during pregnancy. Meanwhile, azithromycin, as an “emerging pollutant”, is increasingly polluting the environment due to the rapidly increasing usage (especially after the COVID-19). Previous studies have suggested a possible teratogenic risk of prenatal azithromycin exposure (PAzE), but its effects on fetal multi-organ development are still unclear. This study aimed to explore the potential impacts of PAzE. Materials and methodsWe focused on pregnancy outcomes, maternal/fetal serum phenotypes, and fetal multiple organ development in mice at different doses (50/200 mg/kg·d) during late pregnancy or at 200 mg/kg·d during different stages (mid-/late-pregnancy) and courses (single-/multi-course). Key findingsThe results showed PAzE increased the rate of the absorbed fetus during mid-pregnancy and increased the intrauterine growth retardation rate (IUGR) during late pregnancy. PAzE caused multiple blood phenotypic changes in maternal and fetal mice, among which the number and degree of changes in fetal blood indicators were more significant. Moreover, PAzE inhibited long bone/cartilage development and adrenal steroid synthesis, promoting hepatic lipid production and ovarian steroid synthesis in varying degrees. The order of severity might be bone/cartilage > liver > gonads > other organs. PAzE-induced multi-organ alterations differed in stages, courses doses and fetal sex. The most apparent changes might be in high-dose, mid-pregnancy, multi-course, and female, while there was no typical rule for a dose-response relationship. SignificanceThis study confirmed PAzE could cause fetal developmental abnormalities and multi-organ functional alterations, which deepens the comprehensive understanding of azithromycin's fetal developmental toxicity.

Prenatal immobility stress: Relationship with oxidative stress, inflammation, apoptosis, and intrauterine growth restriction in rats.

Prenatal stress is a significant risk factor affecting pregnant women and fetal health. In the present study, we aimed to investigate the effect of immobility stress at different periods of pregnancy on oxidative stress, inflammation, placental apoptosis and intrauterine growth retardation in rats. Fifty adult virgin female Wistar albino rats were used. Pregnant rats were exposed to 6 h/day immobilization stress in a wire cage at different stages of pregnancy. Groups I and II (Day 1-10 stress group) were sacrificed on the 10th day of pregnancy, and Group III, Group IV (10-19th-day stress group), and Group V (1-19th-day stress group) were sacrificed on the 19th day of pregnancy. Inflammatory cytokines, including interleukin-6 (IL-6) and interleukin-10 (IL-10), serum corticotropin-releasing hormone (CRH), and corticosterone levels were measured by enzyme-linked immunosorbent assay. Malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT) levels in the placenta were spectrophotometrically measured. Histopathological analyses of the placenta were evaluated by hematoxylin and eosin staining. Tumor necrosis factor-alpha (TNF-α) and caspase-3 immunoreactivity in placenta tissues were determined by the indirect immunohistochemical method. Placental apoptosis was determined by the TUNEL staining method. We found that the immobility stress during pregnancy significantly increased serum corticosterone levels. Our results showed that the immobility stress diminished the number and weight of fetuses in rats compared to the non-stress group. The immobility stress caused significant histopathological changes in the connection zone and labyrinth zone and increased placental TNF-α and caspase-3 immunoreactivity and placental apoptosis. In addition, immobility stress significantly increased the levels of pro-inflammatory IL-6 and MDA and caused a significant decrease in the levels of antioxidant enzymes such as SOD, CAT, and anti-inflammatory IL-10. Our data suggest that immobility stress causes intrauterine growth retardation by activating the hypothalamic-pituitary-adrenal axis and deteriorating placental histomorphology and deregulating inflammatory and oxidative processes.

RETROSPECTIVE DIAGNOSIS OF IMMUNOREGULATORY DISORDERS IN CHILDREN REQUIRING DISPENSARY MONITORING

Objective. To find the relationship between the quantitative indicators of DNA TREC and KREC with three groups of pathological conditions: sudden death syndrome, frequent diseases of the bronchopulmonary system and allergic diseases. Materials and methods. The study was conducted retrospectively. The survey group included infants and young children with various diseases, in which, based on the anamnesis, the presence of a pathology of the immune system was assumed. A total of 43 children were examined, of which group I - 5 children with sudden death syndrome, group II - 25 patients with frequent diseases of the bronchopulmonary system who received long-term antibiotic therapy, group III - 13 children with allergic diseases. Quantitative determination of TREC and KREC was carried out from samples of "dry spots" of blood obtained during neonatal screening. Results. Low TREC values were recorded more frequently than low KREC values, found in 79.1% and 48.8%, respectively. In the group of patients with sudden death syndrome (I group), who died in the first - fourth months of life, there were high rates of intrauterine growth retardation, dysembryogenesis stigma, congenital malformations, intrauterine infections. This group of children is characterized by thymomegaly and hypoplasia of the adrenal glands, TREC and KREC values are reduced in all children. The group of children with a high frequency of bronchopulmonary diseases (Group II) is characterized by a persistent course of herpes virus infection, prolonged subfebrile condition, neutropenia, transient hypoimmunoglobulinemia A, and thymomegaly. Low TREC values were found in 88% of children of group II, KREC - in 32%. In the III group of patients with allergic diseases (bronchial asthma, atopic dermatitis, allergic rhinitis, food allergy), TREC values are reduced in 53.8% of children, KREC – in 61.5% of children. Children of this group often suffered ARI, laryngotracheitis, obstructive bronchitis. During the first months of life, 75% of children manifested atopic dermatitis. In 62.5% of children with allergic diseases, a low level of serum immunoglobulin A was detected. Conclusions. Low values of TREC and KREC in a significant part of the examined patients make it possible to suspect the pathology of the immune system. The presence of diseases belonging to the group of warnings for the presence of primary immunodeficiency may also indicate the pathology of the immune system, primary or transient, due to infections, prematurity, intrauterine growth retardation, toxic effects. Comparison of the results of the survey in the neonatal period and diseases formed in infancy and early childhood was carried out retrospectively. The introduction in the Russian Federation of neonatal screening of primary immunodeficiencies based on the quantitative determination of TREC and KREC makes it possible to timely diagnose immunity errors and prescribe modern methods of treatment. However, the cut-off of the TREC and KREC values needs to be revised in the future.

Lactobacillus derived from breast milk facilitates intestinal development in IUGR rats.

The intestinal microbiota contributes to infant's intestine homeostasis. This study aimed to analyse how probiotics derived from breast milk promote infant intestinal development in rat pups. The effect of potential probiotics derived from breast milk on development of intrauterine growth retardation (IUGR) newborn rats' intestine was investigated. Limosilactobacillus oris ML-329 and Lacticaseibacillus paracasei ML-446 exhibited good hydrophobicity percentages (p < 0.05). ML-446 showed a significant effect on intestinal length and weight (p < 0.05). Meanwhile, the villus height of the IUGR newborn rats fed with ML-329 was significantly higher compared with those fed with Lacticaseibacillus rhamnosus GG (p < 0.05). Moreover, ML-329 and ML-446 both significantly stimulated the proliferation and differentiation of intestinal epithelial cells by increasing the number of ki67-positive cells, goblet cells, and lysozyme-positive Paneth cells (p < 0.05) through Wnt and Notch pathway. The proliferation and differentiation stimulating effects of ML-329 and ML-446 on IECs in the jejunum, ileum, and colon were mediated by activating the Wnt pathway with increased expression of wnt, lrp5, and β-catenin genes and accumulation of β-catenin, and by downregulating the Notch signalling pathway with decreased expression of the activated notch protein. Lactobacillus could facilitate IUGR rat pups' intestinal development and enhance the proliferation of Paneth cells and goblet cells. These findings provide further insights into promotion of the intestinal development by breast milk-derived beneficial microbes in early life of the IUGR newborn rats.

Early supplementation of folate and vitamin B12 improves insulin resistance in intrauterine growth retardation rats.

Insulin sensitivity is changed during the neonatal period in small for gestational age (SGA) infants. Yet, the interventional strategies are still limited. We aimed to investigate the effects of supplementation with high folate and vitamin B12 diets in the early postnatal period on the changes in insulin sensitivity in an intrauterine growth retardation (IUGR) rat model. IUGR rat model was established by both low-protein diet feeding and daily diet restriction. High folate and vitamin B12 diet was supplied in IUGR as nutritional interventional group (IUGR-I), otherwise, the non-intervened IUGR group (IUGR-NI). In this study, male rats were studied in order to avoid hormonal and gender influence. At 21, 60 and 120 days, fasting plasma glucose, insulin, triglyceride, cholesterol, and homocysteine levels were measured among the control, IUGR-I, and IUGR-NI groups. Pearson analysis was used to evaluate the correlation between homocysteine and fasting blood glucose, insulin, HOMA-IR, triglyceride, and cholesterol levels. We established IUGR rat model by both low protein and restricted diet feeding during pregnancy and the incidence of IUGR pups was 93.33%. There was no difference in fasting glucose, insulin, HOMA-IR, triglyceride and cholesterol levels between the control, the IUGR-NI and the IUGR-I group at day 21. At day 60, insulin, HOMA-IR and triglyceride levels in the IUGR-I group were remarkably lower than those in the IUGR-NI group, but still higher than those in the control group (F=38.34, P=0.02; F=49.48, P=0.02; F=17.93, P<0.001, respectively). At day 120, glucose, insulin, HOMA-IR and Hcy levels in the IUGR-I group were obviously lower than those in the IUGR-NI group, although still higher than those in the control group (F=21.60, P<0.001; F=164.46, P<0.001; F=75.15, P<0.001; F=35.46, P<0.001, respectively). There were no significant differences in triglyceride and cholesterol levels between the IUGR-I group and the control group at day 120. At 120-day, homocysteine in IUGR-I group was highly positively correlated with fasting glucose and HOMA-IR (r=0.863, P=0.006; r=0.725, P=0.042, respectively); Only homocysteine was positively correlated with fasting glucose in IUGR-NI group (r=0.721, P=0.044). Early supplementation of folate and vitamin B12 improved insulin resistance and lipid levels in IUGR rats to some extent, along with decreasing homocysteine levels, but not enough to completely repair glucose and lipid metabolism.

SİSTEMİK LUPUS ERİTEMATOZUSLU GEBE KADINLARIN MATERNAL VE FETAL SONUÇLARININ DEĞERLENDİRİLMESİ

Aim: To evaluate obstetrical and perinatal outcomes of pregnancies with systemic lupus erythematosus (SLE).&#x0D; Materials and Method: This was a retrospective study evaluating obstetric outcomes of 35 patients with SLE who were followed up between 2010 and 2020. Lupus activity was based on SLE Disease Activity Index (SLEDAI) criteria and pregnancy outcomes were evaluated by grouping as active and inactive SLE.&#x0D; Results: The mean maternal age was 29 (21-39). The rate of antiphospholipid syndrome and active disease was 5% and 40% respectively. There were 30 live births. Two elective abortions were reported, one with major cardiac anomaly and the other with renal agenesis. One spontaneous abortion and 2 stillbirths were observed in the active SLE group. Overall rates of intrauterine growth retardation, preeclampsia, and preterm delivery were 8%, 20%, and 26%, respectively. Fetal loss and preterm delivery were significantly higher in the active SLE group (p=0,018, p=0,023). A higher rate of neonatal intensive care unit (NICU) admission was observed in the active SLE group (p=0,034) but Apgar scores less than 0,05).&#x0D; Conclusion: Despite advances in the medical sciences, an active disease with SLE has an elevated risk of inducing adverse pregnancy outcomes. Close follow-up could helps reduce pregnancy complications, especially preterm deliveries.

P-gp expression inhibition mediates placental glucocorticoid barrier opening and fetal weight loss

BackgroundPrenatal adverse environments can cause fetal intrauterine growth retardation (IUGR) and higher susceptibility to multiple diseases after birth, related to multi-organ development programming changes mediated by intrauterine overexposure to maternal glucocorticoids. As a glucocorticoid barrier, P-glycoprotein (P-gp) is highly expressed in placental syncytiotrophoblasts; however, the effect of P-gp on the occurrence of IUGR remains unclear.MethodsHuman placenta and fetal cord blood samples of IUGR fetuses were collected, and the related indexes were detected. Pregnant Wistar rats were administered with 30 mg/kg·d (low dose) and 120 mg/kg·d (high dose) caffeine from gestational day (GD) 9 to 20 to construct the rat IUGR model. Pregnant mice were administered with caffeine (120 mg/kg·d) separately or combined with sodium ferulate (50 mg/kg·d) from gestational day GD 9 to 18 to confirm the intervention target on fetal weight loss caused by prenatal caffeine exposure (PCE). The fetal serum/placental corticosterone level, placental P-gp expression, and related indicator changes were analyzed. In vitro, primary human trophoblasts and BeWo cells were used to confirm the effect of caffeine on P-gp and its mechanism.ResultsThe placental P-gp expression was significantly reduced, but the umbilical cord blood cortisol level was increased in clinical samples of the IUGR neonates, which were positively and negatively correlated with the neonatal birth weight, respectively. Meanwhile, in the PCE-induced IUGR rat model, the placental P-gp expression of IUGR rats was decreased while the corticosterone levels of the placentas/fetal blood were increased, which were positively and negatively correlated with the decreased placental/fetal weights, respectively. Combined with the PCE-induced IUGR rat model, in vitro caffeine-treated placental trophoblasts, we confirmed that caffeine decreased the histone acetylation and expression of P-gp via RYR/JNK/YB-1/P300 pathway, which inhibited placental and fetal development. We further demonstrated that P-gp inducer sodium ferulate could reverse the inhibitory effect of caffeine on the fetal body/placental weight. Finally, clinical specimens and other animal models of IUGR also confirmed that the JNK/YB-1 pathway is a co-regulatory mechanism of P-gp expression inhibition, among which the expression of YB-1 is the most stable. Therefore, we proposed that YB-1 could be used as the potential early warning target for the opening of the placental glucocorticoid barrier, the occurrence of IUGR, and the susceptibility of a variety of diseases.ConclusionsThis study, for the first time, clarified the critical role and epigenetic regulation mechanism of P-gp in mediating the opening mechanism of the placental glucocorticoid barrier, providing a novel idea for exploring the early warning, prevention, and treatment strategies of IUGR.

Impact of preimplantation genetic testing on obstetric and neonatal outcomes: a systematic review and meta-analysis

To investigate whether preimplantation genetic testing (PGT) increases the risk of adverse obstetric and neonatal outcomes. Systematic review and meta-analysis. Not applicable. Pregnancies achieved after PGT or invitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI). Systematic search of databases until December 2020 with cross-checking of references from relevant articles in English. Obstetric and neonatal outcomes after PGT and IVF/ICSI, including mean birth weight, low birth weight, very low birth weight (VLBW), mean gestational age at birth, preterm birth, very preterm birth, birth defects, intrauterine growth retardation (IUGR), sex ratio, cesarean section, hypertensive disorders of pregnancy, gestational diabetes mellitus, placenta disorder (placenta previa, placenta abruption, placenta accreta), and preterm premature rupture of membranes. Ultimately, a total of 785,445 participants were enrolled in this meta-analysis, and these participants were divided into a PGT group (n = 54,294) and an IVF/ICSI group (n = 731,151). The PGT pregnancies had lower rates of low birth weight (risk ratio [RR] 0.85, 95% confidence interval [CI] 0.75 to 0.98), VLBW (RR 0.52, 95% CI 0.33 to 0.81), and very preterm births (RR 0.55, 95% CI 0.42 to 0.70) than those of IVF/ICSI pregnancies. However, the PGT group had a higher rate of the obstetric outcome of hypertensive disorders of pregnancy (RR 1.30, 95% CI 1.08 to 1.57). The PGT did not increase the risk of other adverse obstetric and neonatal outcomes, such as those associated with mean birth weight, mean gestational age at birth, birth defects, IUGR, sex ratio, cesarean section, gestational diabetes mellitus, placental disorder (placenta previa, placenta abruption, placenta accreta), or preterm premature rupture of membranes. We performed subgroup analysis with only blastocyst biopsies and found that PGT with blastocyst biopsies was associated with a lower rate of VLBW (RR 0.55, 95% CI 0.31 to 0.95). The PGT with blastocyst biopsies did not increase the risk of other adverse obstetric and neonatal outcomes. Additionally, we performed subgroup analysis with only frozen-thawed embryo transfer cycles, and we found that PGT pregnancies were associated with a lower rate of VLBW (RR 0.55, 95% CI 0.31 to 0.97), a lower rate of cesarean birth (RR 0.90, 95% CI 0.82 to 0.99), a higher rate of preterm birth (RR 1.10, 95% CI 1.02 to 1.18), and a higher rate of IUGR (RR 1.21, 95% CI 1.06 to 1.38) than those of IVF/ICSI pregnancies. The PGT with frozen-thawed embryo transfer did not increase the risk of other adverse obstetric and neonatal outcomes. The pooled analysis suggested that PGT did not increase the risk of adverse obstetric outcomes. The association between PGT and a higher risk of IUGR requires further investigation.

H19/let-7 axis mediates caffeine exposure during pregnancy induced adrenal dysfunction and its multi-generation inheritance

Previously, we systemically confirmed that prenatal caffeine exposure (PCE) could cause intrauterine growth retardation (IUGR) and adrenal steroid synthesis dysfunction in offspring rats. However, the multi-generation inheritance of adrenal dysfunction and its epigenetic mechanism has not been reported. In this study, the PCE rat model was established, part of the pregnant rats were executed on gestational day 20, while the others were delivered normally and the fetal rats were reared into adulthood. The PCE female rats of filial generation 1 (F1) were mated with wild males to produce F2 offspring, and the same way to produce F3 offspring. All the adult female rats of three generations were sacrificed for the related detection. Results showed that PCE could decrease fetal weight, increase IUGR rate, and elevate serum corticosterone level. Meanwhile, the expression of fetal adrenal GR, DNMT3a/3b, miRNA let-7c increased while those of CTCF, H19, and StAR decreased, and the total methylation rate of the H19 promoter region was enhanced. We used SW-13 cells to clarify the molecular mechanism and found that cortisol-induced in vitro changes of these indexes were consistent with those in vivo. We confirmed that high level of cortisol through activating GR, on the one hand, promoted let-7 expression and inhibited StAR expression; on the other hand, caused high methylation and low expression of H19 by down-regulating CTCF and up-regulating DNMT3a/3b, then enhanced let-7 inhibitory effect on StAR by “molecular sponge” effect. Finally, in vivo experiments showed that the adrenal steroid synthesis function and H19/let-7 axis presented the glucocorticoid-dependent changes in the adult female F1, F2, and F3. In conclusion, PCE can cause female adrenal dysfunction with matrilineal multi-generation inheritance, which is related to the programming alteration of the H19/let-7 axis. This study provides a novel perspective to explain the multi-generation inheritance of fetal-originated disease in IUGR offspring.

Docosahexaenoic acid activates the Nrf2 signaling pathway to alleviate impairment of spleen cellular immunity in intrauterine growth restricted rat pups.

Intrauterine growth retardation (IUGR) impairs immune function in children. IUGR is associated with an imbalance of oxidative stress and abnormal apoptosis. Therefore, an IUGR rats model was established to determine the antioxidant capacity and apoptosis in newborn IUGR rats and explored whether these effects were regulated after Docosahexaenoic acid (DHA) supplementation to rat pups. First, eight normal-birth-weight (NBW) and eight IUGR neonatal rats (a 10% low-protein diet) were used to obtain the antioxidant capacity and apoptosis in IUGR rat pups. Then, 32 newborn rats were randomly assigned to the normal birth weight (NBW), DHA supplementation for NBW (ND), IUGR, and DHA supplementation for IUGR (ID) groups. Starting from the 7th day after birth, DHA was given to the experimental group and the same volume of distilled water was given to the control group for 21 days. (1) DHA improved the serum and spleen CD4/CD8 ratios and IL-4 and IFN-γ mRNA expression. (2) DHA decreased the level of MDA, but increased T-AOC in serum and spleen. (3) DHA increased the protein expression of Bcl-2 while decreased Bax. (4) DHA increased protein expression of the Nrf2 signaling pathway and the downstream antioxidant genes GSH-PX and CAT. DHA may alleviate the impairment of spleen cellular immunity in IUGR rat pups by inhibiting oxidative stress and apoptosis related to the activation of Nrf2 signaling pathway.

H3K9ac of TGF\u03b2RI in human umbilical cord: a potential biomarker for evaluating cartilage differentiation and susceptibility to osteoarthritis via a two-step strategy

BackgroundEpidemiological investigation and our previous reports indicated that osteoarthritis had a fetal origin and was closely associated with intrauterine growth retardation (IUGR). Human Wharton’s jelly-derived mesenchymal stem cells (WJ-MSCs) could be programmable to “remember” early-life stimuli. Here, we aimed to explore an early-warning biomarker of fetal-originated adult osteoarthritis in the WJ-MSCs.MethodsFirstly, two kinds of WJ-MSCs were applied to evaluate their chondrogenic potential in vitro through inducing chondrogenic differentiation as the first step of our strategy, one from newborns with IUGR and the other from normal newborns but treated with excessive cortisol during differentiation to simulate the excessive maternal glucocorticoid in the IUGR newborns. As for the second step of the strategy, the differentiated WJ-MSCs were treated with interleukin 1β (IL-1β) to mimic the susceptibility to osteoarthritis. Then, the expression and histone acetylation levels of transforming growth factor β (TGFβ) signaling pathway and the expression of histone deacetylases (HDACs) were quantified, with or without cortisol receptor inhibitor RU486, or HDAC4 inhibitor LMK235. Secondly, the histone acetylation and expression levels of TGFβRI were further detected in rat cartilage and human umbilical cord from IUGR individuals.ResultsGlycosaminoglycan content and the expression levels of chondrogenic genes were decreased in the WJ-MSCs from IUGR, and the expression levels of chondrogenic genes were further reduced after IL-1β treatment, while the expression levels of catabolic factors were increased. Then, serum cortisol level from IUGR individuals was found increased, and similar changes were observed in normal WJ-MSCs treated with excessive cortisol. Moreover, the decreased histone 3 lysine 9 acetylation (H3K9ac) level of TGFβRI and its expression were observed in IUGR-derived WJ-MSCs and normal WJ-MSCs treated with excessive cortisol, which could be abolished by RU486 and LMK235. At last, the decreased H3K9ac level of TGFβRI and its expression were further confirmed in the cartilage of IUGR rat offspring and human umbilical cords from IUGR newborn.ConclusionsWJ-MSCs from IUGR individuals displayed a poor capacity of chondrogenic differentiation and an increased susceptibility to osteoarthritis-like phenotype, which was attributed to the decreased H3K9ac level of TGFβRI and its expression induced by high cortisol through GR/HDAC4. The H3K9ac of TGFβRI in human umbilical cord could be a potential early-warning biomarker for predicting neonatal cartilage dysplasia and osteoarthritis susceptibility.

Dexamethasone-induced intrauterine growth restriction modulates expression of placental vascular growth factors and fetal and placental growth.

Prenatal exposure to glucocorticoids (GC) is a central topic of interest in medicine since GCs are essential for the maturation of fetal organs and intrauterine growth. Synthetic glucocorticoids, which are used in obstetric practice, exert beneficial effects on the fetus, but have also been reported to lead to intrauterine growth retardation (IUGR). In this study, a model of growth restriction in mice was established through maternal administration of dexamethasone during late gestation. We hypothesised that GC overexposure may adversely affect placental angiogenesis and fetal and placental growth. Female BALB/c mice were randomly assigned to control or dexamethasone treatment, either left to give birth or euthanised on days 15, 16, 17 and 18 of gestation followed by collection of maternal and fetal tissue. The IUGR rate increased to 100% in the dexamethasone group (8 mg/kg body weight on gestational days 14 and 15) and pups had clinical features of symmetrical IUGR at birth. Dexamethasone administration significantly decreased maternal body weight gain and serum corticosterone levels. Moreover, prenatal dexamethasone treatment not only induced fetal growth retardation but also decreased placental weight. In IUGR placentas, VEGFA protein levels and mRNA expression of VEGF receptors were reduced and NOS activity was lower. Maternal dexamethasone administration also reduced placental expression of the GC receptor, αGR. We demonstrated that maternal dexamethasone administration causes fetal and placental growth restriction. Furthermore, we propose that the growth retardation induced by prenatal GC overexposure may be caused, at least partially, by an altered placental angiogenic profile.

Pregnancy rates and perinatal outcomes in women with systemic lupus erythematosus: data from the Korean national health claims database.

The pregnancy rate in systemic lupus erythematosus (SLE) is not fully understood and comparisons of adverse pregnancy outcomes (APOs) with SLE versus the general population are limited. This study aimed to estimate the pregnancy rate and APOs in Korean SLE compared to those without SLE. Pregnant women were identified using the ICD-10 codes for delivery and abortion in the Korean national health claims database (2013-2015). APOs were classified as fetal loss, intrauterine growth retardation (IUGR), pre-eclampsia/eclampsia, and gestational diabetes. Annual incidence rates (IRs) of pregnancy and APOs were calculated in women with SLE and the general population without SLE and the two groups were compared using age-adjusted incidence rate ratios (IRRs). Age-stratified IRRs were further analyzed. The annual IRs of pregnancy in SLE were 29.54-30.70 per 1000 persons. The IRRs were lower in women with SLE than in the general population: 0.68 (0.61-0.76), 0.66 (0.60-0.74), and 0.74 (0.66-0.82) in each respective year. The IRRs of fetal loss, IUGR, and pre-eclampsia/eclampsia were 1.30 (1.14-1.49), 4.65 (3.55-6.09), and 3.43 (2.70-4.36), respectively. However, the IRR of gestational diabetes in SLE did not significantly differ from that of women without SLE. Among the APOs, fetal loss, IUGR, and pre-eclampsia/eclampsia showed decreasing tendencies as age increased. Pregnancy rates in SLE were approximately 30% lower than those in the general population. Except for gestational diabetes, fetal loss, IUGR, and pre-eclampsia/eclampsia were higher in SLE and showed a decreasing tendency with age. Key Points • This population-based cohort study showed that pregnancy rates in SLE were approximately 30% lower than those in the general population. • SLE had a 1.3-fold higher rate of fetal loss, more than 4-fold higher IUGR rate, and more than 3-fold pre-eclampsia or eclampsia rate compared with the general population. • Adverse pregnancy outcomes in SLE showed a decreasing tendency with age.

The Melatonin and Enriched Environment Ameliorated Low Protein-Induced Intrauterine Growth Retardation by IGF-1 And mtor Signaling Pathway and Autophagy Inhibition in Rats.

Sprague-Dawley rats were randomly allocated to 3 groups: control (C), model (M) and EE+MEL group. Animals were housed in an enriched environment (EE+MEL group) or remained in a standard environment (C group, M group). IUGR rat model was built by feeding a low protein diet during pregnancy. MEL was administered by gavaging. At day 1 post-birth, the baseline characteristics and serum biochemical parameters, morphology of liver and small intestine, enzyme activities, and mRNA expression levels of fetal rats were determined. The autophagy marker LC3 and Beclin1 were determined by western blot analysis. EE+MEL markedly improved the baseline characteristics, hepatic and intestinal morphology of IUGR fetuses. In addition, the lactase activities in the fetal intestine were markedly increased by the EE+MEL. The levels of serum somatostatin (SST), Growth hormone (GH), GH releasing hormone (GHRH), Insulin-like Growth Factor 1 (IGF-1), triiodothyronine (T3), and tetraiodothyronine (T4) were found to be recovered by EE+MEL. In addition, the EE+MEL significantly ameliorated the mRNA expression of SST, GHRH, and GHRH receptor (GHRHR), GH, GHR, IGF-1, and IGF-1 receptor (IGF1R), IGF binding protein-1 (IGFBP1), mammalian target of rapamycin (mTOR), S6 kinase 1 (S6K1) and eukaryotic initiation factor 4E (eIF4E)-binding protein 1 (4EBP1) in fetuses. In IUGR fetal livers, LC3 and Beclin1 were found to be increased at birth, while LC3 and Beclin1 were observed to be significantly decreased in the EE+MEL group. EE+MEL could improve fetal rats' baseline characteristics, serum biochemical parameters, birth weight, intestinal and hepatic morphology and enzyme activities. These effects could be explained by the activation of the IGF-1/IGFBP1 and IGF-1/mTOR/S6K1/4EBP1 signaling pathway and autophagy inhibition.

Altered Transcription Factor Binding and Gene Bivalency in Islets of Intrauterine Growth Retarded Rats.

Intrauterine growth retardation (IUGR), which induces epigenetic modifications and permanent changes in gene expression, has been associated with the development of type 2 diabetes. Using a rat model of IUGR, we performed ChIP-Seq to identify and map genome-wide histone modifications and gene dysregulation in islets from 2- and 10-week rats. IUGR induced significant changes in the enrichment of H3K4me3, H3K27me3, and H3K27Ac marks in both 2-wk and 10-wk islets, which were correlated with expression changes of multiple genes critical for islet function in IUGR islets. ChIP-Seq analysis showed that IUGR-induced histone mark changes were enriched at critical transcription factor binding motifs, such as C/EBPs, Ets1, Bcl6, Thrb, Ebf1, Sox9, and Mitf. These transcription factors were also identified as top upstream regulators in our previously published transcriptome study. In addition, our ChIP-seq data revealed more than 1000 potential bivalent genes as identified by enrichment of both H3K4me3 and H3K27me3. The poised state of many potential bivalent genes was altered by IUGR, particularly Acod1, Fgf21, Serpina11, Cdh16, Lrrc27, and Lrrc66, key islet genes. Collectively, our findings suggest alterations of histone modification in key transcription factors and genes that may contribute to long-term gene dysregulation and an abnormal islet phenotype in IUGR rats.

2048-P: DNA Methylation Changes Reveal Distal Regulatory Regions in Islets of Intrauterine Growth Restricted Rats

Intrauterine growth retardation (IUGR) is associated with development of type 2 diabetes in adulthood. Using a rat model of IUGR induced by bilateral uterine artery ligation, we performed RNA-Seq and HELP-TagSeq, a restriction enzyme-based genome-wide approach, to assess the association of DNA methylation changes in islets at 2 weeks of age, prior to the onset of diabetes. We identified over 2,000 transcripts differentially expressed in IUGR (q&amp;lt;0.05 vs. control) and 512 differentially methylated regions (DMRs-&amp;gt;3 contiguous CpGs within 500 bp) mapped to 481 genes (q &amp;lt;0.05). 23 and 489 loci showed significantly increased and decreased DNA methylation in IUGR islets, respectively (q &amp;lt;0.05). Integrating methylome and transcriptome datasets showed alterations in multiple genes and canonical pathways that are critical for normal islet function. DMRs were enriched with transcription factor binding motifs, such as Elk1, Etv1, Foxa1, Foxa2, Pax7, Stat3, Hnf1, and AR. These transcription factors were also identified as top upstream regulators by Ingenuity Pathway Analysis of islet transcriptomes. To determine if identified DMRs represented regulatory regions for non-proximal genes, we performed in silico analysis of 3D chromosomal interactions between conserved DMRs and interacting regions of the genome using published human pancreas and islet Hi-C datasets. Hi-C analysis identified interactions between 14 highly conserved DMRs and 35 genes (key islet genes) with expression changes (q &amp;lt; 0.05). Changes in gene expression for many of these persisted until adulthood. In adulthood, there were far more HiC interaction genes associated with altered expression and most of these were key islet genes. Our findings have identified novel DMRs with potential distal regulatory functions, and suggest alterations of DNA methylation in key transcription factor binding motifs and genes in early life that may contribute to gene dysregulation and an abnormal islet phenotype in IUGR rats. Disclosure Y. Lien: None. X.M. Lu: None. R.A. Simmons: None. Funding National Institutes of Health (DK114054)

Serum metabolic profile characteristics of offspring rats before and after birth caused by prenatal caffeine exposure

Epidemiological investigations have confirmed that prenatal caffeine intake could increase the incidence rate of intrauterine growth retardation (IUGR) and multiple diseases after birth. Based on liquid chromatography-mass spectrometry, we analyzed serum metabolic profiles of offspring rats before and after birth in IUGR model induced by prenatal caffeine exposure (PCE). We discovered that differential metabolites in PCE fetuses mainly manifested as amino acids and lipid metabolism. In adulthood, PCE offspring showed less and inconsistent types of differential metabolites compared to those in utero, which still exhibited gender differences. The main differential metabolites induced by PCE, including phospholipids, platelet-activating factor, arachidonic acid, bile acid, sphingosine-1-phosphoric acid, indoxyl sulfuric acid, and cortexolone, may participate in the pathological and physiological processes of organ toxicities. This study demonstrated the short- and long-term developmental toxicity and gender differences of caffeine, providing new ideas for exploring the early warning and drug intervention targets of IUGR offspring.

2188-P: Altered Transcription Factor Binding and Gene Bivalency May Contribute to Long-Term Gene Dysregulation in Intrauterine Growth-Retarded Rat Islets

Intrauterine growth retardation (IUGR), which induces epigenetic modifications and permanent changes in gene expression, has been associated with the development of type 2 diabetes. Using a rat model of IUGR induced by bilateral uterine artery ligation, we performed ChIP-Seq and RNA-Seq to assess the association of genome-wide histone modifications and gene dysregulation in islets from 2 and 10 week rats. IUGR induced significant changes in enrichment of H3K4me3 and H3K27me3 marks in both 2-week and 10-week islets, which were correlated with expression changes of multiple genes critical for normal islet function in IUGR islets. HOMER analysis showed that IUGR-induced histone mark changes were enriched at transcription factor binding motifs, such as Nkx2-1, Nkx2-5, FoxL2, Pax7, Sox4, Mitf, and Oct4-Sox2-Nanog. These transcription factors were also identified by Ingenuity Pathway Analysis of islet transcriptomes as top upstream regulators that regulate the changes in gene expression and biological activities. In addition, our ChIP-seq data revealed more than 1,000 potential bivalent genes as identified by enrichment of both H3K4me3 and H3K27me3. No expression or very low expression of these genes was consistent with a poised state of transcription. The poised state of many potential bivalent genes was altered by IUGR. Interestingly, 3 genes important for islet function, Acod1, FGF-21, and Serpina11, which were poised in 2-week control islets, lost their poised state in 10-week control islets, but gained bivalency in 10-week IUGR islets. In contrast, another 3 genes, Cdh16, Lrrc27, and Lrrc66, which were not poised in control 2-week islets, gained de novo bivalency in 10-week control islets, but lost their poised state in 10-week IUGR islets. Collectively, our findings suggest alterations of histone modification in key transcription factors and genes which may contribute to long-term gene dysregulation and an abnormal islet phenotype in IUGR rats. Disclosure Y. Lien: None. X.M. Lu: None. Z. Wang: None. R.A. Simmons: None. Funding National Institutes of Health

Reduction of the PI3K/Akt related signaling activities in skeletal muscle tissues involves insulin resistance in intrauterine growth restriction rats with catch-up growth.

Individuals with intrauterine growth retardation (IUGR) are prone to insulin resistance, whereas the underlying molecular mechanisms remain unclear. In this study, we investigated if the PI3K/Akt signaling pathway in skeletal muscle tissues involves insulin resistance in IUGR offsprings, particularly ones with catch-up growth. An IUGR rat model was established by feeding rats an isocaloric (30.50 Kcal/g) diet containing 8% protein (low-protein diet) from day 1 of pregnancy until the birth of their pups. Glucometabolic parameters were measured and compared. Quantitative PCR and western blot were performed to assess mRNA and protein expression changes of the PI3K/Akt related signals in skeletal muscle tissues. Pearson analysis was used to assess the correlation of the PI3K/Akt signaling level and catch-up growth with the insulin resistance index (IRI). The values of fasting plasma glucose, fasting insulin and IRI were significantly higher, whereas insulin sensitivity index was significantly lower in IUGR offsprings than those in the controls. The PI3K mRNA and protein levels as well as the phospho-AktSer473 levels were significantly lower in IUGR offsprings compared to the controls. Reductions of GLUT4 as well as increases of PTEN and nuclear fractional PPARγ were detected in IUGR offsprings. Catch-up growth IUGR rats were positively correlated with insulin resistance and underwent more remarkable alterations of the PI3K, PTEN and GLUT4 expressions. Our results demonstrated that rats born IUGR developed insulin resistance later in life, which was likely mediated by reductions of the PI3K/Akt related signaling activities, particularly in those with excess catch-up growth.

The burden to be second twin: a population-based study of 2686 twins: (2124 dichorionic). Proposal of the concept of mobility

Objectives: To identify if there is a specific neonatal morbidity/mortality among second twins relative to first twins.Study design: A 17-year (2001–2017) population-based observational cohort of all twin newborns born in the South of Reunion island after 21 weeks.Results: Among 1062 dichorionic (DTP) and 281 monochorionic twin pregnancies (2686 newborns), twin 2 have a doubled risk to be in breech presentation and a bad Apgar at 1 mn (≤6) in vaginal deliveries. Specific to dichorionic pregnancies, twin 2 were lighter by 50–60 g than twin 1, had higher rates of intrauterine growth retardation (IUGR), OR 1.33, p = .007, a doubled risk to have congenital abnormalities OR 2.1, p = .006.Conclusion: In dichorionic twin pregnancies, second twins having a doubled prevalence of severe congenital abnormalities are not completely elucidated and deserves further research. (1) We propose that twin 2 presenting higher risks of being IUGR and much higher risks of severe malformations suggest that during pregnancy, the less mobile of the two twins is “relegated” to the back of the uterus. (2) For interventions in the delivery room, systematically the most experimented neonatologist should plan to manage the second twin because significantly twin 2 presents higher problems than twin 1.