Apoptosis in an Ovine Model of Intrauterine Growth Restriction (IUGR)

子宫内生长受限 (IUGR) 绵羊模型中的细胞凋亡

• 批准号: 8484312
• 负责人: JUAN A ARROYO
• 金额: $ 23万
• 依托单位: BRIGHAM YOUNG UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-05-01 至 2014-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8484312
• 关键词: Address; Affect; Animals; Apoptosis; Apoptosis Regulator; Apoptotic; BAX gene; BCL1 Oncogene; BIRC4 gene; Bax protein; Caspase; Cells; Cleaved cell; Cytokeratin 18; DNA; DNA Nucleotidylexotransferase; Data; Development; Disease; Event; Family; Fetal Development; Fetal Growth; Fetal Growth Retardation; Fever; Functional disorder; Goals; Growth; Growth and Development function; HSPB1 gene; Heating; Human; Hypoxia; Hypoxia Inducible Factor; In Vitro; Induced Hyperthermia; Infant Mortality; Instruction; Label; Lead; Mediating; Mediator of activation protein; Modeling; Morbidity - disease rate; Mus; Nutrient; Oxygen; Phase; Placenta; Placental Insufficiency; Placentation; Pregnancy; Protein Family; Proteins; Protocols documentation; Reporting; Role; Sheep; Stress; Techniques; Telomerase; Testing; Vascular remodeling; Villus; base; caspase-3; caspase-9; human disease; hyperthermia treatment; in vivo; insight; pro-apoptotic protein; response; transcription factor; trophoblast

Intrauterine growth restriction (lUGR) is a significant cause of increased infant mortality and morbidity. A number of placental abnormalities have been described during lUGR including an increase in trophoblast apoptosis. Abnormal trophoblast apoptosis may lead to impaired oxygen and nutrient exchange, suggesting a role for apoptosis in the development of lUGR. To better understand apoptosis in lUGR pregnancies we will utilize an established hyperthermia sheep lUGR model which mimics the human disease considerably. We propose to look at apoptosis in the sheep placentae early in gestation and in sheep exposed to hyperthermia for 20 days, 55 days when placental growth is at its peak and 80 days near term when fetal growth is maximal. Our long-term goals are: 1) to determine the in-vivo placental apoptotic effects, DNA degradation, proliferation rate, telomerase activity and cytokeratin 18 cleavage in our model of hyperthermia induced PI-IUGR in the sheep, 2) to elucidate the expression anti-apoptotic molecules BCL-2 and XIAP and the pro-apoptotic molecules BAX, caspase 3 and caspase 9 in the sheep PI-IUGR placentae. To determine the correlation and expression of HSP27 and of HIF-2a during apoptosis in hyperthermia induced PI-IUGR, and 3) to determine the effect of hyperthermia, hypoxia or both on apoptosis, cytokeratin 18 cleavage, telomerase activity and expression of BCL-2, XIAP, BAX, caspase 3 and 9, HSP27 and HIF-2a in cultured placental binucleated cells (BNC) obtained from normal sheep placentomes. In order to accomplish these we will determine apoptosis using the terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling technique (TUNEL) and cytokeratin 18 cleavage expression. Also we will determine the telomerase activity associated with our model of lUGR using the Telomerase Repeat Amplification Protocol (TRAP) technique. To further study apoptosis, we will determine the protein levels of Bcl-2, Bax, and caspase 3 and 9 activation in placentae from animals treated with hyperthermia as compared to controls. Also, we will determine the effects of hyperthermia in the expression of XIAP and HIF-2a in the placentae from treated animals. To study apoptosis in-vitro we will determine the effects of hyperthermia and hypoxia on cultured placental BNCs.

宫内生长受限（IUGR）是婴儿死亡率和发病率增加的重要原因。一 在lUGR期间已经描述了许多胎盘异常，包括滋养层细胞增多， 凋亡异常的滋养层细胞凋亡可能导致氧和营养交换受损，提示 凋亡在IUGR发展中的作用。为了更好地了解lUGR妊娠中的细胞凋亡， 将利用已建立的热疗绵羊IUGR模型，该模型相当大地模拟了人类疾病。 我们建议观察妊娠早期绵羊胎盘和暴露于 热疗20天，胎盘生长高峰期55天，胎儿足月80天， 增长最大。我们的长期目标是：1）确定体内胎盘凋亡效应，DNA 细胞角蛋白18的降解、增殖率、端粒酶活性和切割 诱导绵羊PI-IUGR，2）阐明抗凋亡分子BCL-2和XIAP的表达， 促凋亡分子BAX、caspase 3和caspase 9在绵羊PI-IUGR胎盘中的表达。以确定 HSP 27和HIF-2a在高温诱导的PI-IUGR中凋亡过程中的表达及相关性， 和3）确定高温、缺氧或两者对细胞凋亡，细胞角蛋白18裂解， 端粒酶活性及BCL-2、XIAP、BAX、caspase 3和9、HSP 27和HIF-2a表达 从正常绵羊胎盘获得的胎盘双核细胞（BNC）。为了实现这些目标，我们 将使用末端脱氧核苷酸转移酶介导的dUTP缺口末端标记来确定细胞凋亡 TUNEL法和细胞角蛋白18切割表达。同时检测端粒酶活性 与我们的使用端粒酶重复扩增方案（TRAP）技术的lUGR模型相关。 为了进一步研究细胞凋亡，我们将测定Bcl-2、Bax和caspase 3和9活化的蛋白水平。 与对照组相比，用高温处理的动物的胎盘中。此外，我们将确定 高温对处理动物胎盘中XIAP和HIF-2a表达的影响。研究 体外细胞凋亡我们将确定高温和缺氧对培养的胎盘BNC的影响。