Endocan expression is increased in the placenta from obese women with gestational diabetes mellitus

IntroductionEndocan, a member of the proteoglycan family, is involved in a wide range of diseases including obesity and diabetes. The aim of this study was to determine the effect of (i) maternal obesity and gestational diabetes mellitus (GDM) on placental endocan expression; and (ii) endocan knockdown on markers of inflammation.MethodsEndocan mRNA and protein expression was determined in human placenta from (i) lean and obese and normal glucose tolerant (NGT) pregnant women (n = 10 patients per group); and (ii) women with GDM and BMI-matched NGT women (n = 40 patients per group). Primary villous trophoblast cells and HUVECs were used to assess the effect of endocan siRNA knockdown on pro-inflammatory cytokines.ResultsThere was no effect of maternal obesity on placental endocan expression. Further, endocan expression was not different between lean NGT and BMI-matched women with GDM. However, endocan mRNA and protein expression was significantly higher in placenta from obese women with GDM when compared to BMI-matched NGT women. Knockdown of endocan in villous trophoblast cells and HUVECs had no effect on infection- or inflammation-induced expression and secretion of IL-6, IL-8 and MCP-1.DiscussionEndocan expression is increased in the human placenta from obese women with GDM, and in response to pro-inflammatory stimuli. Loss-of-function studies in villous trophoblast cells and HUVECs revealed that endocan is not directly involved in the genesis or in the expression of pro-inflammatory cytokines induced by LPS or IL-1β. Further studies are required to elucidate the functional consequences of increased placental endocan expression in obese GDM pregnancies.     

Placental glucose transporter (GLUT)-1 is down-regulated in preeclampsia

IntroductionTransplacental fetal glucose supply is predominantly regulated by glucose transporter-1 (GLUT1). Altered expression and/or function of GLUT1 may affect the intrauterine environment, which could compromise fetal development and may contribute to fetal programming. To date it is unknown whether placental GLUT1 is affected by preeclampsia, which is often associated with intrauterine growth restriction (IUGR). We addressed the hypothesis that preeclampsia leads to decreased expression and function of placental GLUT1.MethodsPlacentae were obtained following normal pregnancy and from pregnancies affected by preeclampsia. Washed villous tissue fragments were used to prepare syncytial microvillous (MVM) and basal plasma membranes (BM) microvesicles. GLUT1 protein and mRNA expression was assessed by western blot analysis and qPCR using Fast SYBR Green. A radio-labeled glucose up-take assay using placenta-derived syncytial microvesicles was used to analyze GLUT1 function.ResultsGLUT1 protein expression was significantly down-regulated in (apical) MVM of the syncytiotrophoblast in preeclampsia (n = 6) compared to controls (n = 6) (0.40 ± 0.04 versus 1.00 ± 0.06, arbitrary units, P < 0.001, Student's t-test), while GLUT1 mRNA expression did not show a significant difference. In addition, the functional assay in syncytial microvesicles showed a significantly decreased glucose transport activity in preeclampsia (61.78 ± 6.48%, P < 0.05) compared to controls. BM GLUT1 protein expression was unchanged and glucose up-take into BM microvesicles showed no differences between the preeclampsia and control groups.DiscussionOur study shows for the first time that in preeclampsia placental GLUT1 expression and function are down-regulated at the apical plasma membrane of the syncytiotrophoblast. Further studies are needed to assess whether these changes occur also in vivo and contribute to the development of IUGR in preeclampsia.     

Maternal BMI and gestational diabetes alter placental lipid transporters and fatty acid composition

IntroductionPlacental fatty acid (FA) uptake and metabolism depend on maternal supply which may be altered when women have a high pre-pregnancy body mass index (BMI) or develop gestational diabetes (GDM). Consequently, an impaired FA transport to the fetus may negatively affect fetal development. While placental adaptation of maternal-fetal glucose transfer in mild GDM has been described, knowledge on placental FA acid metabolism and possible adaptations in response to maternal obesity or GDM is lacking.We aimed to analyze the FA composition and the expression of key genes involved in FA uptake and metabolism in placentas from women with pre-pregnancy normal weight (18.5 ≤ BMI<25 kg/m2), overweight (25 ≤ BMI<30 kg/m²), obesity (BMI ≥ 30 kg/m²), and lean pregnant women with GDM.MethodsPlacental FA content was determined by gas liquid chromatography. Placental mRNA expression of FA transport proteins (FATP1, FATP4, FATP6), FA binding proteins (FABP3, FABP4, FABP7), FA translocase (FAT/CD36) and enzymes (Endothelial lipase (EL) and lipoprotein lipase (LPL)) were quantified by qRT-PCR.ResultsHigh pre-pregnancy BMI and GDM were associated with decreased placental FATP1, FATP4, EL and increased FAT/CD36 and FATP6 expressions. LPL mRNA levels and placental total FA content were similar among groups. Specific FA, including some long-chain polyunsaturated FA, were altered.DiscussionOur results demonstrate that high pre-pregnancy BMI or GDM independently alter mRNA expression levels of genes involved in FA uptake and metabolism and the placental FA profile, which could affect fetal development and long-term health.     

Effect of selective fetectomy on morphology of the mouse placenta

IntroductionPlacental examination is recommended when genetic mutations cause fetal lethality in mice. But how fetal death alters histomorphology of the surviving mouse placenta is not known.MethodsPlacentas were examined at E17.5 after fetectomy of 1–2 fetal mice per pregnancy at either embryonic day (E) 15.5 (N = 8; Fx-2 group) or E13.5 (N = 5; Fx-4 group), which left 12 ± 2 surviving fetuses per litter.ResultsFetectomy caused no changes in placental weights and no increases in placental hypoxia (pimonidazole staining). The size and cell morphology of the decidua and junctional zone regions were unchanged and, in the Fx-2 group, these regions became significantly less hypoxic. Significant changes in labyrinth volume included a 30% increase in the Fx-2 group and in both groups, a >50% decrease in % fetal blood space and >40% increase in % labyrinth tissue. Maternal blood sinusoid volume was unchanged. Cell death in the labyrinth was significantly increased (22-fold increase in TUNEL staining) whereas placental mRNA expression of the proliferation marker Mki67 was unchanged. mRNA expression of sFlt1 and Prl3b1 (mPL-II) was unchanged in the labyrinth and junctional zone tissues in the Fx-2 group and in whole placental tissue in the Fx-4 group.DiscussionPlacental examination of the junctional zone and decidual regions after spontaneous fetal death in late gestation is likely to yield useful phenotypic information and abnormalities that may contribute to fetal death. In contrast, labyrinth abnormalities including increased tissue volume and reduced fetoplacental vascularity may not be due to genetic perturbation nor predate fetal death.     

Activation of the IL-1β/CXCL1/MMP-10 axis in chorioamnionitis induced by inactivated Group B Streptococcus

Infection or inflammation during pregnancy is known to lead to maternal immune activation triggering a fetal inflammatory response syndrome associated with deleterious effects, such as brain injury and neurodevelopmental disabilities. Group B Streptococcus (GBS) - one of the most common bacterium colonizing pregnant women - can be responsible for chorioamnionitis. Given that interleukin (IL)-1β has a major role in anti-GBS host defense, we hypothesized that IL-1β-driven innate immune response is implicated in GBS-induced chorioamnionitis. Using a rat model of GBS-induced chorioamnionitis, this study showed that inflammatory response to this pathogen was associated with maternal and placental IL-1β hyper expression. Following placental chemokine (C-X-C motif) ligand 1 (CXCL1) production, polymorphonuclear leukocytes (PMN) placental infiltration started at 24 h post-GBS exposure, and MMP-10 was released within these placentas. At 72 h, PMN infiltration extended to membranes and to membranes' arteries. This was associated with IL-1β release within the fetus blood at 72 h. Such a GBS-associated inflammatory cascade might be deleterious for fetal organs. These results pave the way toward targeted placento-protective anti-inflammatory strategies against GBS-induced chorioamnionitis.     

Placental Pim-1 expression is increased in obesity and regulates cytokine- and toll-like receptor-mediated inflammation

IntroductionObesity is a growing epidemic, and as a consequence the number of obese pregnancies has also increased. Pregnancy is characterised by maternal and placental inflammation which is intensified with maternal obesity. The proviral integration site for Moloney murine leukemic virus (Pim)-1 protein is a serine/threonine kinase involved in a wide range of inflammatory diseases. In relation to obesity, however, its role has not been elucidated in human placenta. The aims were to determine the placental expression of Pim-1 with pre-existing maternal obesity and its role in regulating placental inflammation associated with obesity.MethodsHuman placenta was obtained at the time of term Caesarean section from lean and pre-existing obese pregnant women to determine the effect of maternal obesity on Pim-1 expression. To determine the effect of Pim-1 on the inflammatory response induced by bacterial endotoxin LPS and pro-inflammatory cytokines TNF-α or IL-1β, the chemical inhibitor SMI-4a and siRNA were used.ResultsPim-1 protein and mRNA expression was significantly increased in placenta of obese women. SMI-4a significantly suppressed the expression and release of pro-inflammatory cytokine IL-6 and chemokines GRO-α and MCP-1 when stimulated with LPS or TNF-α in placenta. Primary trophoblast cells transfected with Pim-1 siRNA had decreased expression and release of pro-inflammatory cytokines IL-1β, IL-6, chemokines GRO-α and MCP-1, when stimulated with LPS, TNF-α or IL-1β.DiscussionThe findings from this study implicate Pim-1 may contribute to placental inflammation in pregnancies complicated by maternal obesity. Thus, therapeutic targets for Pim-1 may improve fetal outcomes complicated by obese pregnancies.     

Neuronal apoptosis in the neonates born to preeclamptic mothers

Abstract

Objective: Preeclampsia may result in uteroplacental insufficiency and chronic intrauterine fetal distress. The aim of this study is to address this issue investigating neuronal apoptosis in an experimental model of preeclampsia and to evaluate the neurological outcome of the perinatal asphyxia in the neonates born to preeclamptic mother.

Materials and methods: Two out of four pregnant Sprague–Dawley rats (preeclamptic group) were given water containing 1.8% NaCl on gestation day 15 and 22 in order to establish the model of preeclampsia whereas other two (non-preeclamptic group) received normal diet. A model of perinatal asphyxia was established on the postnatal 7th day to one preeclamptic and one non-preeclamptic dam. Overall 23 pups born to overall four dams were decapitated to assess neuronal apoptosis by the TUNEL assay.

Results: The number of apoptotic neuronal cells was significantly higher in the preeclampsia groups in comparison with the control group (p?=?0.006 and p?=?0.006, respectively). It was also significantly higher in the asphyctic/non-preeclamptic group than the count in the control group (p?=?0.01). There was also significant difference between both asphyctic groups (p?=?0.003).

Conclusion: We conclude that preeclampsia causes small babies for the gestational age and cerebral hypoplasia. Both preeclampsia and perinatal asphyxia can cause increased neuronal apoptosis in the neonatal brains. However, the prognosis for neurological outcome is much worse when the perinatal asphyxia occurs in newborns born to preeclamptic mothers.     

Placenta-bound and Body Fluid PP13 and its mRNA in Normal Pregnancy Compared to Preeclampsia,HELLP and Preterm Delivery

ObjectivesTo compare the distribution of placental protein 13 (PP13) in fetal and maternal blood and amnionic fluid and to correlate it with PP13 protein and mRNA in the placenta.MethodsUmbilical arterial serum, amnionic fluid, maternal venous serum and placental tissues were collected from normal outcome pregnancies (N = 63) (GA>37), early onset preeclampsia (PE) (N = 12, GA: 26–33), and HELLP syndrome (N = 5, GA: 27–29). Because PE and HELLP cases delivered preterm, cases of preterm delivery (PTD) (N = 6, GA: 31–36) served as additional control. PP13 was determined by ELISA, Western blot, and immunohistochemistry. PP13 mRNA was measured by PCR (RT-PCR). Continuous parameters were compared by t-test, P < 0.05 was considered significant.ResultsIn women with normal pregnancy outcome significantly higher PP13 levels were found in maternal serum compared to amnionic fluid and negligible amount was found in fetal serum. A similar pattern was identified in cases of PTD with concentrations similar to term control. In PE and HELLP cases PP13 levels in amnionic fluid level were more than twice compared to maternal serum (P < 0.001). Umbilical cord level was negligible in PE but high in HELLP corresponding to the much higher level of PP13 in this patient group compared to all others. In the placenta PP13 level in term controls was higher compared to PTD. In PE and HELLP (similar early delivery time as PTD) the level was significantly higher (P < 0.01) compared to PTD or term controls. PP13 mRNA levels in term control and PTD were similar while PP13 mRNA levels in PE and HELLP placentas were significantly lower compared to term controls or PTD or the two combined. Syncytiotrophoblast labeling appeared stronger in PE and HELLP compared to term controls and PTD.ConclusionsIn all cases but HELLP, PP13 in fetal blood is very low indicating that routing of PP13 to fetal blood is limited and that the fetus is unlikely to generate PP13. PP13 mRNA is lower in the third trimester at the time of disease while protein level accumulates and become higher creating an unparallel change in the level of the mRNA and the corresponding protein.     

Dual In Vitro Perfusion of an Isolated Cotyledon as a Model to Study the Implication of Changes in the Third Trimester Placenta on Preeclampsia

In the current study perfusions of an isolated cotyledon of term placenta using standard medium were compared to medium containing xanthine plus xanthine oxidase (X + XO), which generates reactive oxygen species (ROS). A time-dependant increase in the levels of different cytokines (TNF-α, IL-1ß, IL-6, IL-8 and IL-10) was observed between 1 and 7 h with more than 90% of the total recovered from the maternal compartment with no significant difference between the 2 groups. For 8-iso-PGF2α 90% of the total was found in the fetal compartment and a significantly higher total release was seen in the X + XO group.Microparticles (MPs) isolated from the maternal circuit were identified by flow cytometry as trophoblastic sheddings, whereas MPs from the fetal circuit were predominantly derived from endothelial cells. More than 90% of the total of MPs was found in the maternal circuit. The absolute amount of the total as well as the maternal fraction were significantly higher in the X + XO group.Immunohistochemistry (IHC) of the perfused tissue revealed staining for IL-1β of villous stroma cells, which became clearly more pronounced in experiments with X + XO. Western blot of tissue homogenate revealed 2 isoforms of IL-1β at 17 and 31 kD. In X + XO experiments there was a tendency for increased expression of antioxidant enzymes in the tissue.Western blot of MPs from the maternal circuit showed increased expression of antioxidant enzymes in the X + XO group and for IL-1β only the 17 kD band was detected.In vitro reperfusion of human placental tissue results in mild tissue injury suggestive of oxidative stress. In view of the increased generation of ROS in perfused tissue with further increase under the influence of X + XO, the overall manifestation of oxidative stress remained rather mild. Preservation of antioxidant capacity of human placental tissue could be a sign of integrity of structure and function being maintained in vitro by dual perfusion of an isolated cotyledon. The observed changes resemble findings seen in placentae from preeclampsia.     

Expressions of apoptosis-regulating factors in bovine retained placenta

The aim of the present study was to evaluate the relationship between the retention of fetal membranes (RFM) and apoptosis of the cells in fetal membranes. The present study investigated mRNA and protein expressions of apoptosis-regulating factors: FAS, cellular FLICE-like inhibiting protein (cFLIP), BAX, BCL2, caspase-8 (CASP8), and CASP3 in fetal membranes. Placentomes were manually collected from the uterus immediately after parturition and classified into two groups (RFM; n = 8 and non-RFM; n = 8) according to whether placental membranes were expelled or not within 12 h after delivery. FAS mRNA expression in maternal placental tissue was less in RFM cows than in non-RFM cows (P < 0.05). cFLIP mRNA expression in maternal and fetal placental tissue was greater in RFM cows than in non-RFM cows (P < 0.05). CASP3 mRNA expression in maternal placental tissue was greater in RFM cows than in non-RFM cows (P < 0.05). However, the protein expressions of FAS, cFLIP and cleaved CASP3 were not significantly different between the two groups. mRNA and protein expressions of BAX, BCL2 and CASP8 were also not significantly different between the two groups. In the immunohistochemical study, single-stranded DNA, which appears specifically in the apoptotic cells, was mainly found in the maternal placenta of non-RFM cows. Together these results suggest that RFM occurs at least in part due to a dysfunctional apoptotic process caused by the inhibition of FAS expression in the maternal placenta, and the increase of cFLIP expression in the maternal and fetal placenta.     

The NR4A receptors Nurr1 and Nur77 are increased in human placenta from women with gestational diabetes

IntroductionMembers of the NR4A subfamily are involved in a wide range of diseases including obesity and diabetes. The aim of this study was to determine the effect of maternal obesity and gestational diabetes mellitus (GDM) on the expression of the NR4A receptors Nurr1, Nur77 and NOR1.MethodsHuman placenta was obtained at the time of term Caesarean section from (i) lean and obese and normal glucose tolerant (NGT) pregnant women; and (ii) women with GDM and BMI-matched NGT women (n = 16 patients). Primary trophoblast cells, isolated from human term placenta, were used to determine the effect of pro-inflammatory cytokines on NR4A protein expression. Primary trophoblast cells were also used to assess the effect of Nurr1, Nur77 and NOR1 siRNA knockdown on pro-inflammatory cytokines.ResultsThere was no effect of pre-existing maternal obesity on Nurr1, Nur77 or NOR1 expression. Nurr1 and Nur77 expression were significantly higher in GDM placenta compared to NGT placenta, and in the presence of the pro-inflammatory cytokines TNF-α and IL-β in primary trophoblast cells. Knockdown of Nurr1 and Nur77 in human primary trophoblast cells significantly decreased TNF-α induced expression and secretion of IL-6 and IL-8.DiscussionNurr1 and Nur77, which were increased in human placenta from women with GDM, are involved in TNF-α induced-expression of pro-inflammatory cytokines. Pro-inflammatory cytokines are known to play a role in placental nutrient transport. Thus, the regulation of pro-inflammatory cytokines by Nurr1 and Nur77 suggest that these proteins may play a role in placental function and transport mechanisms.     

Differential effect of LPS and IL-1β in term placental explants

IntroductionInflammation is an important cause of placental dysfunction often associated with pregnancy complications. One well-known cause of inflammation is infection, through conserved “pathogen-associated molecular patterns” (PAMPs). Endogenous inducers of inflammation, known as “damage-associated molecular patterns” (DAMPs), have also been associated with pathological pregnancies and could contribute to the observed placental inflammation. Although both stimuli (i.e. PAMPs/DAMPs) can induce inflammation, they have yet to be studied together to compare their inflammatory effects on the placenta.MethodsWe used a model of term placental explants to compare the effects of a classical PAMP, bacterial lipopolysaccharide (LPS), and a DAMP, the pro-inflammatory cytokine interleukin (IL)-1. Gene and protein expression of several cytokines were analysed by qPCR and ELISAs and immunohistochemistry performed to study placental resident immune cells and apoptosis.ResultsLPS induced pro-inflammatory mediators (IL-6, IL-1β/α, TNF-α) whereas IL-1β induced only IL-6. Furthermore, LPS but not IL-1 exposure, led to elevated IL-10 and IL-1Ra secretion. Blocking the IL-1 signalling pathway abrogated the pro-inflammatory actions of LPS, whilst anti-inflammatory effects were preserved. The number of CD45 ⁺ immune cells was elevated in explants treated with LPS only. A subpopulation of CD45 ⁺ cells were positive for PCNA indicating proliferation of tissue resident macrophages.DiscussionWe conclude that LPS, a classical PAMP, and IL-1, a DAMP, have shared and distinct actions with pro-inflammatory effects mediated through IL-1 but anti-inflammatory actions having a distinct pathway. Identification of an inflammatory mediator (i.e. IL-1) common to multiple stimuli could be a therapeutic target to preserve the placenta.     

Effect of nitric oxide deficiency on tissue-type plasminogen activator expression in the umbilical cord in a pregnancy-induced hypertension rat model

ObjectivesOur study investigates the effects of nitric oxide (NO) deficiency during pregnancy on coagulation and fibrinolysis balance in fetal circulation.Main outcome measuresPregnant rats were treated with or without oral N^G-nitro-l-arginine methyl ester (L-NAME). Systolic blood pressure (SBP) and urinary protein were measured. On gestational day 20, mRNA levels of tissue-type plasminogen activator (tPA), tissue factor (TF), and TF pathway inhibitor (TFPI) in the umbilical cord, placenta, and maternal aorta were evaluated. Immunohistochemical staining of the placenta for PA inhibitor-1 (PAI-1) was performed.ResultsL-NAME treatment in pregnant rats caused significant SBP elevation and severe proteinuria. In the L-NAME-treated group, weights of fetuses and placentae were diminished. tPA mRNA expression decreased in the umbilical cord and placenta, whereas TF and TFPI mRNA levels did not change. Intense PAI-1 immunoreactivity was observed in a part of degenerated placenta. In the aorta, tPA mRNA expression increased in the L-NAME-treated group, while TFPI mRNA levels were lower than in controls.ConclusionsNO deficiency during pregnancy decreased tPA mRNA expression in the umbilical cord and placenta but not in the maternal aorta. Imbalance between coagulation and fibrinolysis in fetal and maternal circulations may, at least in part, contribute to fetal growth restriction.     

Toll-like receptor-2 mediates local innate immune response against Trypanosoma cruzi in ex vivo infected human placental chorionic villi explants

BackgroundCongenital Chagas disease is caused by the protozoan parasite Trypanosoma cruzi that must cross the placental barrier during transmission. The trophoblast constitutes the first tissue in contact with the maternal-blood circulating parasite. Importantly, the congenital transmission rates are low, suggesting the presence of local placental defense mechanisms. On the other hand, the placenta is considered an immune regulatory organ since it acts as a modulator of fetal and maternal immune responses. We have previously proposed that local placental factors, such as the epithelial turnover of the trophoblast and the innate immune response initiated by Toll-like receptors (TLRs), might prevent parasite infection and congenital transmission.Here, we studied in an ex vivo infected human placental chorionic villi explant HPCVE model, the relationship between TLR-2 activation in response to T. cruzi trypomastigotes, the secreted profile of cytokines, the integrity of the placental barrier and the expression of trophoblast turnover markers.ResultsTLR-2 inhibition increases the parasite induced histopathological damage, prevents secretion of IL-6 and IL-10, decreases expression of PCNA (proliferation marker) and of β-hCG (differentiation marker) while increasing caspase 3 activity (cell death marker).ConclusionOur results suggest that TLR-2 is not only involved in the local secretion of cytokines but also regulates, at least partially, the trophoblast turnover.     

细胞因子IL-6mRNA与IL-10mRNA在早产合并绒毛膜羊膜炎胎盘组织中的表达水平及意义

目的:研究白细胞介素-6 mRNA(IL-6 mRNA)和白细胞介素-10 mRNA(IL-10 mRNA)在早产合并绒毛膜羊膜炎孕妇胎盘组织中的表达水平,探讨IL-6及IL-10在妊娠中的作用,以及其在早产合并绒毛膜羊膜炎中的变化及意义。方法:取足月分娩及早产孕妇部分胎盘胎膜组织做病理检查,根据病检结果分为3组:早产感染组20例、早产非感染组20例和足月组20例。采用二步法RT-PCR法测定3组胎盘组织中IL-6 mRNA和IL-10 mRNA的表达。结果:早产感染组胎盘组织中IL-6 mRNA表达水平明显高于早产非感染组和足月组,差异有显著性(P<0.05);而IL-10 mRNA表达水平明显低于另外两组,差异有显著性(P<0.05)。结论:感染可引起细胞因子IL-6 mRNA、IL-10 mRNA在胎盘组织中的表达发生改变,推测IL-6可促进分娩发动,使妊娠提前终止,而IL-10在维持正常妊娠中可能起重要作用。     

A mouse model of term chorioamnionitis: unraveling causes of adverse neurological outcomes

Maternal fever and/or chorioamnionitis at term are associated with an increased prevalence of adverse neurobehavioral outcomes in exposed offspring. Since the mechanisms of such injury are currently unknown, the objectives of this study were to elucidate whether intrauterine inflammation at term results in fetal brain injury. Specifically, we assessed brain injury by investigating the cytokine response, white matter damage, and neuronal injury and viability. A mouse model of intrauterine inflammation at term was utilized by injecting lipopolysaccharide (LPS), or normal saline into uterine horn. Compared to saline-exposed, LPS-exposed fetal brains had significantly increased IL-1β and IL-6 messenger RNA (mRNA) expression (P < .05 for both) and IL-6 protein levels by enzyme-linked immunosorbent assay (ELISA; P < 0.05). Fetal neurons were affected by the intrauterine and fetal brain inflammation, as demonstrated by significantly decreased microtubule-associated protein 2 (MAP2) mRNA expression and a decrease in immunocytochemical staining (a marker of neuronal cytoskeleton development; P < .05), altered neuronal morphology (P < 0.05), and delayed neurotoxicity (P < .05). These fetal neuronal changes occurred without overt changes in white matter damage markers. Marker of astrocyte development and astrogliosis (glial fibrillary acidic protein [GFAP]) did not show an increase; pro-oligodendrocyte marker (PLP1/DM20) was not significantly changed (P > .05). These studies may provide a critical mechanism for the observed long-term adverse neurobehavioral outcomes after exposure to chorioamnionitis at term.     

Evaluation of the use of anti-TNF-α in an LPS-induced murine model

ObjectiveTumor necrosis factor α (TNF-α) may play a critical role in inflammatory-mediated preterm labor. Medications blocking the activity of TNF-α have been shown to be effective in the treatment of conditions such as rheumatoid arthritis; however, the use of these medications for an event like preterm birth or fetal death is unknown. We hypothesized that treatment with anti-TNF-α may decrease the rate of fetal death and preterm birth in a LPS-induced murine model.MethodsPregnant C57BL/6J mice received intraperitoneal (IP) injections of either vehicle or 2 mg anti-TNF-α. After 24 h, 10 μg of LPS was administered IP. Mice were sacrificed 24 h later and outcomes between groups were assessed. A second set of experiments utilizing RT-PCR was performed to determine the influence of anti-TNF-α on production of inflammatory cytokines in response to LPS.ResultsThere were 72 resultant pups in the LPS + saline group, and 91 in the group receiving LPS + anti-TNF-α. Pretreatment with anti-TNF-α reduced the rate of fetal death and preterm birth after LPS administration (p < 0.01). Expression of IL-6, IL-1beta, TLR-2, CD14 and COX-1 were found to be significantly reduced in mice treated with anti-TNF-α and LPS compared to LPS alone.ConclusionThe use of anti-TNF-α decreased fetal deaths and preterm deliveries in an LPS-induced model of preterm birth. In addition, there were critical gene expression alterations in the group receiving anti-TNF-α. Further evaluation of TNF-α blockade as a potential treatment for preterm labor is warranted.     

Pentoxifylline inhibits lipopolysaccharide-induced inflammatory mediators in human second trimester placenta explants

BackgroundIntrauterine infection and inflammation during pregnancy, which leads to up-regulation of inflammatory cytokines and prostaglandin synthesis, has been implicated in the pathogenesis of preterm delivery and other pregnancy complications. Effective preventive and therapeutic strategies to reduce these outcomes are lacking to date. Pentoxifylline (PTX) is a non-specific phosphodiesterase inhibitor which raises intracellular cyclic adenosine monophosphate and decreases production of pro-inflammatory mediators while enhancing anti-inflammatory cytokines. We hypothesized that pentoxifylline will decrease lipopolysaccharide (LPS)-induced pro-inflammatory cytokines production in human placental explants.MethodsPlacental explants derived from normal second trimester human placentas were treated with PTX, stimulated with LPS and cultured at 37 °C in 5% CO₂. Conditioned media were assayed for pro- and anti-inflammatory mediators with multiplex immunoassays or ELISA, and explant tissues for mRNA with real time PCR. Means of PTX-treated and untreated samples were compared using paired t tests and Wilcoxon-signed rank tests.ResultsPTX preferentially inhibited placental expression and production of LPS-induced pro-inflammatory cytokines including TNF-α (25461 vs. 1908 pg/ml, p < 0.001), IL-1β (2921 vs. 1067 pg/ml, p < 0.001) and IFN-γ (2190 vs 427 pg/ml, p < 0.001) with relative preservation of anti-inflammatory mediators. The suppressive effects on LPS-induced placental inflammation were independent of the timing of PTX administration in relation to LPS-induced stimulation.ConclusionOur study suggests that PTX attenuates the LPS-induced pro-inflammatory milieu in human placental explants. We speculate that PTX may have utility as a candidate anti-inflammatory agent for prophylaxis and/or treatment of human placental inflammation.     

Asphyxie périnatale et infirmité motrice d’origine cérébrale (I- Le diagnostic)

Cerebral palsy (CP) is a group of disorders of the development of movement and posture, causing activity limitations, that are attributed to nonprogressing disturbances that occurred in the developing fetal or infant brain. The motor abnormalies are often accompagnied by disturbances of sensation, perception, cognition, behavior and/or by a seizure disorder. The prevalence of CP has not decreased in developed countries over the past 30 years, despite the widespread use of electronic fetal heart rate monitoring and a 5- to 6-fold increase in the cesarean delivery rate. In the term newborn, CP may be attributed to perinatal asphyxia in case of metabolic acidosis in the cord blood (pH < 7,00 and base deficit > 12 mmol/L), followed by a moderate or severe neonatal encephalopathy within 24 hours and a further neurological impairement characterized by spastic quadriplegia and dyskinesia/dystonia. Dating the time of fetal asphyxia during delivery is possible when there are acute catastrophic complications during labor and unexpected acute or progressive fetal heart rate anomalies after a normal admission test, when there is a need for intensive neonatal resuscitation, a multi-organ failure within 72 hours of birth and visualization of acute non focal cerebral abnormalities, mainly by early magnetic resonance imaging (MRI). MRI sequences show either a brain-damaged pattern of the central basal ganglia, thalami and posterior limbs of internal capsules with relative cortical sparing, in acute, near-total asphyxial insults manifested by a continuous bradycardia or a pattern of cortical injury in the watershed zones and relative sparing of the central grey matter, in prolonged partial asphyxia, manifested by late or atypical variable decelerations with progressive fetal tachycardia, loss of reactivity and absent fluctuation. Prolongation of either type of asphyxial insult results in more global brain damage. In order to differentiate a CP occurring after perinatal asphyxia from other neurological sequelae in relation with infection, hemorrhage, stroke, malformations, genetic or metabolic diseases, it is essential that a definitive information from the brain by MRI and an extensive histological examination of the placenta are at disposal.