p38 MAP Kinase and Akt Interaction in HUVEC

HUVEC 中 p38 MAP 激酶和 Akt 相互作用

• 批准号: 7253731
• 负责人: SHEHLA PERVIN
• 金额: $ 7.7万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-06-01 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7253731
• 关键词: 1-Phosphatidylinositol 3-Kinase; Androgen Receptor; Androgens; Apoptosis; Attenuated; Birth Weight; Cell Survival; Cells; Cessation of life; Charcoal; Data; Endothelial Cells; Estradiol; Estrogens; Exposure to; Female; Fetus; Figs - dietary; Growth; Growth Factor; Human; Induction of Apoptosis; Infant; Intracranial Hemorrhages; Investigation; Link; MAP2K6 gene; MAPK14 gene; Measures; Mitogen-Activated Protein Kinases; Molecular; Neonatal; Newborn Respiratory Distress Syndrome; Nitric Oxide; Nutrient; Pathway interactions; Phenols; Placenta; Play; Predisposition; Protein Overexpression; RNA; Rate; Role; SB 203580; Serum; Sex Characteristics; Signal Transduction; Staining method; Stains; Stress; TdT-Mediated dUTP Nick End Labeling Assay; Testing; Transfection; Umbilical cord structure; Umbilical vein; Up-Regulation; Withdrawal; annexin A5; base; caspase-3; cytokine; deprivation; human MAPK14 protein; human female; human male; inhibitor/antagonist; kinase inhibitor; male; mortality; neonate; novel; novel strategies; response; upstream kinase; wortmannin

DESCRIPTION (provided by applicant): Over all objective of this study is to understand the molecular mechanisms and gender differences in response of male and female human umbilical vein endothelial cells (HUVEC) to stress, with emphasis on cross talk between PI-3 Kinase/Akt and p38 MAP Kinase pathways. It is found that male fetus has greater mortality rate than female. The molecular basis of the observed gender difference in mortality of male and female fetus is poorly understood. Phosphatidylinositol 3-kinase (PI-3 Kinase)/ Akt pathway is the key controller of endothelial cell viability determining endothelial cell viability. Withdrawal of growth factor or exposure to cytokines inhibits PI-3 Kinase /Akt signaling in endothelial cells and induces apoptosis. Decline of PI-3 Kinase/Akt induces an upregulation of stress induced p38 MAP Kinase. The cross-talk between mitogen -activated protein kinase (MAP Kinase) and PI-3 Kinase/Akt pathway and the role of activated p38 MAP Kinase in apoptosis is poorly understood. In this novel proposal we link the viability of fetus to the functional integrity and survival of HUVEC which is maintained by PI-3 Kinase /Akt signaling. Although there is lack of textual data, we have included preliminary data where male HUVEC is more susceptible than female to wortmannin (an inhibitor of PI-3 Kinase) induced stress, leading to sharper decline in pAkt and dramatic onset of apoptosis. We will consolidate this preliminary finding by assessing the susceptibilities of male and female HUVECs to a variety of stress. This proposal will also determine whether gender difference in susceptibility of male and female HUVEC to stress is regulated by p38 MAP Kinase. SB 203580 (a specific inhibitor of p38 MAP Kinase) or small-inhibitory RNA (si-RNA) against p38 MAP Kinase will be used to inhibit p38 MAP Kinase activity. Overexpression of MKK6, an upstream kinase of p38 MAP Kinase will also be utilized to elucidate the potential cross-talk between p38 MAP Kinase and PI-3 Kinase/ Akt pathway. Apoptosis induction will be determined by TUNEL assay, Annexin V staining and caspase-3 and -9 activities. We will also examine whether 17-¿-estradiol (E2) modulates the cross talk between p38 MAP Kinase and PI-3 Kinase /Akt signaling in male and female HUVEC. Human umbilical vein endothelial cells from male and female umbilical cord will be used to study the gender difference in the response to stress induced apoptosis. This investigation could help us understand the mechanisms by which the mortality rate of male fetus and neonates is higher than female.

描述（由申请人提供）：本研究的总体目标是了解男性和女性人脐静脉内皮细胞 (HUVEC) 对应激反应的分子机制和性别差异，重点是 PI-3 激酶/Akt 和 p38 MAP 激酶途径之间的交互作用。研究发现，男性胎儿的死亡率高于女性。所观察到的男性和女性胎儿死亡率性别差异的分子基础尚不清楚。磷脂酰肌醇 3-激酶 (PI-3 激酶)/Akt 通路是内皮细胞活力的关键控制器，决定内皮细胞的活力。停止生长因子或暴露于细胞因子会抑制内皮细胞中的 PI-3 激酶/Akt 信号传导并诱导细胞凋亡。 PI-3 激酶/Akt 的下降会导致应激诱导的 p38 MAP 激酶上调。丝裂原激活蛋白激酶 (MAP 激酶) 和 PI-3 激酶/Akt 通路之间的串扰以及激活的 p38 MAP 激酶在细胞凋亡中的作用知之甚少。在这项新颖的提议中，我们将胎儿的活力与 HUVEC 的功能完整性和存活联系起来，而 HUVEC 由 PI-3 激酶/Akt 信号传导维持。尽管缺乏文本数据，但我们已经纳入了初步数据，其中男性 HUVEC 比女性更容易受到渥曼青霉素（PI-3 激酶抑制剂）诱导的应激的影响，导致 pAkt 急剧下降和细胞凋亡的急剧发生。我们将通过评估男性和女性 HUVEC 对各种压力的敏感性来巩固这一初步发现。该提案还将确定男性和女性 HUVEC 对应激的敏感性的性别差异是否受到 p38 MAP 激酶的调节。 SB 203580（p38 MAP 激酶的特异性抑制剂）或针对 p38 MAP 激酶的小抑制性 RNA (si-RNA) 将用于抑制 p38 MAP 激酶活性。 p38 MAP 激酶的上游激酶 MKK6 的过表达也将用于阐明 p38 MAP 激酶和 PI-3 激酶/Akt 通路之间潜在的串扰。细胞凋亡诱导将通过TUNEL测定、膜联蛋白V染色以及caspase-3和-9活性来确定。我们还将检查 17-¿-雌二醇 (E2) 是否调节男性和女性 HUVEC 中 p38 MAP 激酶和 PI-3 激酶/Akt 信号传导之间的串扰。来自男性和女性脐带的人脐静脉内皮细胞将用于研究对应激诱导的细胞凋亡反应的性别差异。这项研究可以帮助我们了解男性胎儿和新生儿死亡率高于女性的机制。