NMR STUDIES OF HFH PROTEINS

该项目计划研究人类胎盘新陈代谢， 特别是能量和碳水化合物的代谢，这是基本的 确保正常胎儿健康所必需的过程。胎盘 在有氧条件下有利于氧化磷酸化，但其 大量的葡萄糖消耗和乳酸产生表明糖酵解是 也是一条重要的途径。每条途径的相对重要性在 正常和异常的胎盘是未知的。我们想测试一下 假设氧化磷酸化和糖酵解都是 正常胎盘中活跃的代谢途径，但异常 胎盘，即糖尿病胎盘，可能有利于糖酵解增加 由于长期暴露在高水平的葡萄糖和局部性 缺氧。糖尿病胎盘能量代谢的改变可能是 与胎盘功能改变相关，导致临床 观察到不良的胎儿结局。胎盘新陈代谢研究将 使用磁共振波谱(MRS)进行观察 动态生化过程是非侵入性的、连续的。夫人 将用于研究灌流装置内的胎盘绒毛。 绒毛悬浮在中间，并明显地维持了几个月 持续氧合、营养灌流、pH和 温度控制。三磷酸腺苷、无机磷和糖的变化 代表正在进行的氧化磷酸化的磷酸盐峰和 将观察糖酵解情况。组织功能将通过以下方式评估 监测葡萄糖和氧气消耗，以及乳酸、乳酸 脱氢酶、雌二醇、孕酮和绒毛膜促性腺激素 放手。我们将描述氧化磷酸化的作用。 以及正常胎盘在基线条件下的糖酵解 缺氧、高血糖、低血糖、酸中毒。对以下问题的回应 将观察到氰化物和碘乙酸酯等抑制剂。我们会 对活组织和高氯酸进行光谱分析 在固定时间产生更好分辨率光谱的组织提取物 积分。在正常和正常情况下建立绒毛行为后 实验条件下，糖尿病胎盘将在 进行了相似条件下的对比分析。我们希望 对正常和糖尿病胎盘能量有更好的了解 和碳水化合物的代谢，使用强大的无创方法 所获得的信息不仅具有潜在的临床应用价值 意义，但也可能是对未来的重要补充 胎盘的体内光谱学研究。

This project proposes to study human placental metabolism, specifically energy and carbohydrate metabolism which are fundamental processes necessary to ensure normal fetal well being. The placenta favors oxidative phosphorylation under aerobic conditions, but its large glucose consumption and lactate production suggest glycolysis is also an important pathway. The relative importance of each pathway in normal and abnomal placentas is unknown. We would like to test the hypothesis that oxidative phosphorylation and glycolysis are both active metabolic pathways in normal placentas but that abnormal placentas, i.e., diabetic placentas, may favor increased glycolysis because of chronic exposure to high levels of glucose and localized hypoxia. Changes in energy metabolism in diabetic placentas may be associated with altered placental functions leading to clinically observed poor fetal outcomes. Placental metabolism studies will be performed using magnetic resonance spectroscopy (MRS) which observes dynamic biochemical processes noninvasively and continuously. MRS will be used to study placental villi within a perifusion apparatus. The villi are suspended in medium and viably maintained for several hours by continuous oxygenation, nutrient perifusion, pH and temperature control. Changes in ATP, inorganic phosphate, and sugar phosphate peaks representing ongoing oxidative phosphorylation and glycolysis will be observed. Tissue function will be evaluated by monitoring glucose and oxygen consumption, and lactate, lactate dehydrogenase, estradiol, progesterone and chorionic gonadotropin release. We will characterize the roles of oxidative phosphorylation and glycolysis in normal placentas under baseline conditions such as hypoxia, hyper and hypoglycemia, and acidosis. Responses to inhibitors such as cyanide and iodoacetate will be observed. We will perform spectroscopy of viable tissue as well as perchloric acid tissue extracts which yield better resolved spectra at fixed time points. After establishing villous behavior under normal and experimental conditions, diabetic placentas will be studied under similar conditions and comparative analysis performed. We hope to acquire a better understanding of normal and diabetic placental energy and carbyhydrate metabolism by using the powerful noninvasive method of MRS. Information gained will not only have potential clinical significance, but may also represent important complements to future in vivo spectroscopic studies of the placenta.