NMR STUDIES OF HFH PROTEINS

本项目拟研究人类胎盘代谢， 特别是能量和碳水化合物代谢， 这是确保胎儿健康所必需的。 胎盘 在有氧条件下有利于氧化磷酸化，但其 大量葡萄糖消耗和乳酸盐产生表明糖酵解 也是一个重要的途径。 每种途径的相对重要性 正常和异常胎盘是未知的。 我们想测试一下 假设氧化磷酸化和糖酵解都是 活跃的代谢途径在正常胎盘，但异常 胎盘，即，糖尿病胎盘，可能有利于增加糖酵解 由于长期暴露于高水平的葡萄糖和局部 缺氧 糖尿病胎盘能量代谢的变化可能是 与胎盘功能改变相关，导致临床上 观察到不良的胎儿结局。 胎盘代谢研究将 使用磁共振波谱（MRS）进行， 动态生化过程的非侵入性和连续性。 夫人 将用于研究灌注装置内的胎盘绒毛。 绒毛悬浮在培养基中， 小时，通过连续充氧、营养灌注、pH和 温度控制 ATP、无机磷酸盐和糖的变化 磷酸盐峰代表正在进行的氧化磷酸化， 将观察糖酵解。 将通过以下方式评价组织功能： 监测葡萄糖和氧消耗，以及乳酸，乳酸 脱氢酶、雌二醇、孕酮和绒毛膜促性腺激素 release. 我们将描述氧化磷酸化的作用 和糖酵解在正常胎盘在基线条件下， 缺氧、高血糖和酸中毒。 Responses to 将观察到抑制剂如氰化物和碘乙酸盐。 我们将 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光谱的组织提取物 点 在正常条件下建立绒毛行为， 在实验条件下，将研究糖尿病胎盘 进行相似条件和比较分析。 我们希望 更好地了解正常和糖尿病胎盘能量 和碳水化合物代谢， 获得的信息不仅具有潜在的临床意义， 重要性，但也可能是对未来的重要补充 胎盘的活体光谱研究。

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.