The Role Of Subclinical Infection And Cytokines In Prete

亚临床感染和细胞因子在 Prete 中的作用

• 批准号: 7334071
• 负责人: ROBERTO ROMERO
• 金额: --
• 依托单位: EUNICE KENNEDY SHRIVER NATIONAL INSTITUTE OF CHILD HEALTH & HUMAN DEVELOPMENT
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7334071
• 关键词: Role; Subclinical; Infection; Cytokines; Prete

Premature birth is the leading cause of perinatal mortality and morbidity worldwide. The Perinatology Research Branch has defined preterm labor as a syndrome and determined that at least 25% of preterm neonates are born to women with sub-clinical intrauterine infection. The Branch has also provided evidence that many premature neonates are critically ill before birth and proposed that, in the context of intrauterine infection, the onset of premature labor has survival value. The goal of this project is to understand the pathophysiology of premature labor and delivery. This year, our research focused on the following studies: 1. Gene expression signature of spontaneous term labor without histologic chorioamnionitis. Human parturition involves a common pathway manifested clinically by uterine contractions, cervical ripening and chorioamniotic membrane/decidual activation, culminating in membrane rupture. Previous studies have linked parturition to an inflammatory process. Investigators at the Branch analyzed both the transcriptome in the chorioamniotic membranes and maternal blood in order to identify the biological processes in normal spontaneous labor using an unbiased genome-wide approach. Transcriptional profiles for chorioamniotic membranes and blood were generated from patients at term with no labor and patients at term in labor. All placentas were subjected to histologic examination, and a criterion for inclusion was the absence of histologic chorioamnionitis. An increased expression of multiple chemokines and transcripts associated with neutrophil and monocyte recruitment was observed among patients in term labor. The results of this study indicate that labor induces gene expression changes consistent with localized inflammation despite the absence of histological chorioamnionitis. 2. Uterine transcriptomes of bacteria-induced and ovariectomy-induced preterm labor (PTL) in mice are characterized by differential expression of arachidonate metabolism genes. PTL is a syndrome with multiple etiologies. In this study, the Branch focused on two well established mechanisms of PTL: intrauterine infection and the requirement of progesterone for pregnancy maintenance. To investigate the role of intrauterine infection and progesterone in human PTL, investigators analyzed the uterine transcriptome of mice using two models: the first was the induction of PTL by intrauterine infection with Escherichia coli, and the second was the induction of PTL by ovariectomy and progesterone withdrawal. Quantitative reverse transcriptase?polymerase chain reaction measurements demonstrated that bacteria-induced PTL substantially increased the expression of genes involved in prostaglandin synthesis. In contrast, ovariectomy-induced PTL increased the expression of genes involved in lipoxin, leukotriene, and hydroxyeicosatetraenoic acid synthesis. The results of this study indicate that bacteria-induced and ovariectomy-induced PTL each express a different balance of genes that are involved in prostaglandin synthesis and the synthesis of lipoxins, leukotrienes, and HETEs. This balance may be important to PTL and is amenable to experimental investigation. 3. The transcriptome of the uterine cervix before and after spontaneous term parturition. Cervical ripening is one of the components of the common pathway for parturition. Therefore, determining the expression profile of genes involved in this process is crucial for the understanding of mechanisms leading to premature cervical ripening and preterm delivery. This study provided an unbiased and comprehensive description of the changes in the cervical transcriptome before and after spontaneous term labor. The transcriptome of cervical tissue from patients at term not in labor and after spontaneous labor were characterized using Affymetrix microarrays. Results of this study revealed that the cervical transcriptome of term patients who underwent labor is dramatically different from that of patients without labor, with 1192 differentially expressed genes. Among the up-regulated genes were those involved in neutrophil chemotaxis, apoptosis, extracellular matrix regulation, and steroid metabolism. Genes involved in neutrophil chemotaxis were up-regulated in specimens from women after spontaneous labor, and an increased expression of IL-8, IL-6, and scular endothelial growth factor was confirmed by real-time quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR). Toll-like receptor (TLR)-3 and TLR-5 showed decreased gene expression in patients after spontaneous labor in the microarray analysis and this was confirmed by qRT-PCR. 4. A rapid bedside test for the prediction of preterm delivery (PTD). Intra-amniotic infection (IAI) and inflammation have been causally linked to preterm labor (PTL), PTD and fetal injury. Intra-amniotic inflammation can be detected by performing an amniocentesis and measuring cytokines (e.g. IL-6 and MMP-8), white blood cell count, and glucose concentrations in the amniotic fluid. It has been previously demonstrated that elevated MMP-8 concentrations in the amniotic fluid of patients with PTL and intact membranes can diagnose intra-amniotic inflammation with 95% sensitivity and 93% specificity. This year, the Branch conducted a study to investigate the diagnostic indices, predictive values, efficiency and likelihood ratios of the MMP-8 PTD Check test for the detection of IAI, inflammation, spontaneous PTD and severe neonatal morbidity among patients with increased uterine contractions and intact membranes. The MMP-8 PTD Check Test (SK Pharma Co, Ltd, Kyunggi-do, Korea) is a rapid bedside test (configured similarly to a rapid pregnancy test) designed to provide a quick bedside assessment for elevated concentrations of MMP-in a small amount of amniotic fluid. A positive MMP-8 rapid test correctly predicted 70% of the spontaneous PTDs within 48 hrs, and 94% of those occurring within 7 days and 14 days of the amniocentesis. The efficiency of a positive MMP-8 rapid test in the identification of IAI and inflammation was 94% (311/331) and 93% (308/331), respectively. We concluded that this rapid test gives clinicians a fast and accurate assessment of the inflammatory status of the amniotic cavity, and allows for better identification of patients at risk for impending PTD. Follow-up clinical trials are required to determine the role of the MMP-8 rapid test for the identification of inflammation at the time of amniocentesis, as well as determine whether, based on the rapid test results, treatment with antibiotics and/or anti-inflammatory agents may improve pregnancy outcome. 5. Phylogenetic analysis reveals the evolution of the mammalian placenta. The placenta is the lifeline for mammalian reproduction and a complex organ that provides clues about natural selection and evolution. Through phylogenetic analysis of molecular and anatomical data, investigators at the Branch presented evidence describing the evolutionary history of the placenta of eutherian mammals?the group that includes all mammal species except marsupials and the egg laying monotremes (for example, the duck billed platypus). In contrast to most theories, the study found that the disc-shaped, hemochorial placenta of many primates, including humans, existed throughout the eutherian lineage from the last common ancestor of placental mammals to the emergence of humans. That intimate contact between fetal and maternal blood was established in the last common ancestor of the crown group of Eutheria gives credence to the hypothesis that successful pregnancy requires appropriate allorecognition and tolerance at the maternal-fetal interface.

早产是全球围产期死亡率和发病率的主要原因。围产科研究部门将早产劳动定义为综合征，并确定至少有25％的早产新生儿是宫内诊所内感染的妇女出生的。该分支机构还提供了证据表明，许多早熟的新生儿在出生之前患了重病，并提出，在宫内感染的背景下，早产劳动的发作具有生存价值。该项目的目的是了解早产和分娩的病理生理学。今年，我们的研究重点是以下研究： 1。无组织学绒毛膜炎的自发性劳动的基因表达特征。人类的分娩涉及一条通用途径，在临床上通过子宫收缩，宫颈成熟和绒毛膜膜膜/骨化激活，最终导致膜破裂。先前的研究已将分娩与炎症过程联系起来。该分支机构的研究人员分析了绒毛膜膜和孕妇血液中的转录组，以便使用无公开的基因组方法来鉴定正常自发劳动的生物学过程。绒毛膜膜膜和血液的转录曲线是由患者在没有劳动的患者中产生的，没有劳动和患者的劳动期。所有胎盘均经过组织学检查，包含的标准是没有组织学绒毛膜炎。在期限内，患者在患者中观察到了与中性粒细胞和单核细胞募集相关的多个趋化因子和转录本的表达增加。这项研究的结果表明，尽管没有组织学绒毛膜炎，但劳动诱导基因表达与局部炎症一致。 2。小鼠中细菌诱导的和卵巢切除术诱导的早产（PTL）的子宫转录组的特征是蛛网膜酸性代谢基因的差异表达。 PTL是具有多种病因的综合征。在这项研究中，该分支集中在PTL的两种良好的机制上：宫内感染和孕激素维持妊娠的要求。为了研究宫内感染和孕酮在人PTL中的作用，研究人员使用两个模型分析了小鼠的子宫转录组：第一个是通过大肠杆菌内宫内宫内感染诱导PTL，第二个是通过卵巢切除术和孕酮戒断来诱导PTL的PTL。定量逆转录酶？聚合酶链反应测量表明，细菌诱导的PTL显着增加了参与前列腺素合成的基因的表达。相反，卵巢切除术诱导的PTL增加了参与脂氧素，白细胞和羟基羟基乙烯烯酸合成的基因的表达。这项研究的结果表明，细菌诱导的和卵巢切除术诱导的PTL每个人都表达了与前列腺素合成有关的基因的不同平衡，以及脂毒素，白细胞和Hetes的合成。这种平衡对PTL可能很重要，并且可以进行实验研究。 3。自发性分娩前后子宫子宫颈的转录组。宫颈成熟是分娩常见途径的组成部分之一。因此，确定此过程中涉及的基因的表达谱对于理解导致过早宫颈成熟和早产的机制至关重要。这项研究对自发性劳动前后宫颈转录组的变化提供了无偏见的全面描述。使用Affymetrix微阵列对患者的宫颈组织的转录组进行了术语和自发劳动后的特征。这项研究的结果表明，接受劳动的术语患者的宫颈转录组与无劳动的患者的宫颈转录组大不相同，有1192个差异表达的基因。在上调的基因中，有参与中性粒细胞趋化性，凋亡，细胞外基质调节和类固醇代谢的基因。自发性劳动后，女性的标本中，中性粒细胞趋化性的基因被上调，IL-8，IL-6的表达增加，而Scular内皮生长因子通过实时定量逆转录酶 - 聚合酶链链反应（QRT-PCR）证实。在微阵列分析中，自发性劳动后，TOLL样受体（TLR）-3和TLR-5显示出患者的基因表达降低，并且通过QRT-PCR证实了这一点。 4。对早产预测（PTD）的快速床边测试。 - 肿瘤内感染（IAI）和炎症与早产（PTL），PTD和胎儿损伤有因果关系。可以通过进行羊膜穿刺术和测量细胞因子（例如IL-6和MMP-8），白细胞计数和羊水中的葡萄糖浓度来检测 - 肿瘤内炎症。以前已经证明，PTL和完整膜患者的羊水中MMP-8浓度升高可以诊断出95％的敏感性和93％的特异性，可以诊断出肿瘤内炎症。今年，该分支机构进行了一项研究，以研究MMP-8 PTD检查测试的诊断指数，预测值，效率和可能性比，以检测IAI，炎症，自发性PTD以及心脏收缩和完整膜增加的患者的新生儿发病率。 MMP-8 PTD检查测试（SK Pharma Co，Ltd，Kyunggi-Do，韩国）是一种快速的床边测试（与快速妊娠试验相似，旨在为少量的羊水液体提供较高浓度的MMP浓度。 MMP-8阳性快速测试正确地预测了48小时内的70％的自发PTD，而在羊膜内穿刺术的7天和14天内发生的PTD和94％发生。 MMP-8阳性快速测试在IAI和炎症鉴定中的效率分别为94％（311/331）和93％（308/331）。我们得出的结论是，这项快速测试使临床医生可以快速准确评估羊水腔的炎症状态，并可以更好地识别有即将来临的PTD风险的患者。需要进行后续临床试验，以确定MMP-8快速测试在羊膜穿刺时鉴定炎症的作用，并确定基于快速测试结果，使用抗生素和/或抗炎药的治疗是否可以改善妊娠结局。 5。系统发育分析揭示了哺乳动物胎盘的演变。胎盘是哺乳动物繁殖的生命线和一个复杂的器官，可提供有关自然选择和进化的线索。通过对分子和解剖学数据的系统发育分析，该分支的研究者提供了描述Eutherian哺乳动物胎盘的进化史的证据？包括所有哺乳动物物种的组，除了有袋动物和卵单卵单抗以外的所有哺乳动物（例如，鸭子的platypus）。与大多数理论相反，研究发现，包括人类在内的许多灵长类动物的椎间盘形，下的胎盘胎盘都存在于整个Eutherian血统，从胎盘哺乳动物的最后一个共同祖先到人类的出现。胎儿血液与母亲血液之间的亲密接触是在冠冕王室群体的最后一个共同祖先中建立的，证明了成功的怀孕需要在母亲界面上进行适当的同种识别和容忍度的假设。