MECHANISMS OF ACTIONS OF ALCOHOL AND OMEGA3 FATTY ACIDS

申请人摘要：血浆载脂蛋白E(ApoE)是一种糖蛋白 在“反向胆固醇运输”(RCT)中起重要作用的是什么？ 高密度脂蛋白的功能，即(一)从外周组织中去除胆固醇 和/或(Ii)将其输送到肝脏。值得注意的是，乙醇减少了 血浆高密度脂蛋白-载脂蛋白E，而omega3-脂肪酸阻止了这种下降。 我们的初步工作表明，人类血浆高密度脂蛋白-载脂蛋白E也降低了 酗酒者。我们进一步表明，慢性乙醇抑制肝脏 ApoE唾液酸化。载脂蛋白E唾液酸缺乏可能影响其 与高密度脂蛋白相关。反过来，从高密度脂蛋白中失去载脂蛋白E可能会损害其 RCT功能。因此，确认载脂蛋白E是否 高密度脂蛋白的浓度和载脂蛋白E的唾液酸含量 然后展示这些变化是如何影响高密度脂蛋白代谢和 功能。这项研究的90%将利用人类，只有其余的 将使用大鼠：人类研究：载脂蛋白E和高密度脂蛋白将来自人类血清 酗酒者、非酗酒者和非酗酒者。具体问题 载脂蛋白E中唾液酸缺乏的程度是否影响(I) 载脂蛋白E与高密度脂蛋白的关系及(Ii)载脂蛋白E与高密度脂蛋白的解离 高密度脂蛋白-载脂蛋白E与胆固醇的关系。三、载脂蛋白E的损失 来自高密度脂蛋白影响其从外周血中清除胆固醇的能力 纸巾？与非酗酒者相比，人类酗酒者是否有所减少： 高密度脂蛋白中载脂蛋白E的浓度？五、高密度脂蛋白-载脂蛋白E中唾液酸含量 高密度脂蛋白从外周组织中去除胆固醇的能力 人类巨噬细胞？vii.高密度脂蛋白将胆固醇输送到肝脏的能力 人HepG2肝细胞大鼠研究：载脂蛋白E唾液酸化是否在 以上是硬件描述语言的重要方面，同样重要的是确定 肝脏载脂蛋白E的翻译后修饰。这可以 只能在老鼠这样的实验动物身上进行。同样，如果omega3- 脂肪酸确实纠正了乙醇引起的上述缺陷，它更简单 而且测试它们对高密度脂蛋白-载脂蛋白E功能的影响所需的时间更短 老鼠比人类还多。因此，提出的问题是什么是 乙醇和欧米茄3-脂肪酸对VIII.的影响 载脂蛋白E在亚细胞水平的翻译后修饰？IX. 抑制肝唾液酸转移酶活性的机制是什么？ 具体地说，乙醇是否抑制唾液酸转移酶的合成？ 并通过下调其肝脏的mRNA水平？

APPLICANT'S ABSTRACT: Plasma apolipoprotein E (apoE) is a glycoprotein which plays important roles in "Reverse Cholesterol Transport" (RCT) functions of HDL, viz., (i) cholesterol removal from peripheral tissues and/or (ii) its delivery to the liver. Significantly, ethanol decreased plasma HDL apoE, whereas omega3-fatty acids prevented this decrease. Our preliminary work shows that plasma HDL apoE is also reduced in human alcoholics. We have further shown that chronic ethanol inhibits hepatic sialation of apoE. Sialic acid deficiency in apoE may affect its association with HDL. In turn, the loss of apoE from HDL may impair its RCT functions. Therefore, it is important to confirm whether apoE concentration in HDL and sialic acid content of apoE are reduced in alcoholics and then show how these changes affect HDL metabolism and functions. 90% of the study will utilize humans and only the remainder will use rats:Human Study: ApoE and HDL will be from sera of human alcoholics and non-alcoholics and non-alcoholics. The specific questions are: Does the degree of sialic acid deficiency in apoE affect (I) the association of apoE with HDL and (II) dissociation of apoE from HDL & association of HDL- apoE with cholesterol. III. does the loss of apoE from HDL affect its ability to remove cholesterol from peripheral tissues? Do human alcoholics compared to non-alcoholics have decreased: IV. concentration of apoE in HDL? V. sialic acid content of HDL-apoE? VI. HDL ability to remove cholesterol from the peripheral tissues using human macrophages?VII. HDL ability to deliver cholesterol to liver using the human HepG2 liver cells?Rat Study: If apoE sialation is crucial in the above important aspects of HDL it is equally important to determine the post-translational modifications of apoE in the liver. This can only be done in experimental animals like rats.Similarly, if omega3- fatty acids do correct the above defects caused by ethanol it is simpler and less time consuming to test their effects on HDL-apoE functions in rats than in humans. Thus, the questions asked are what are the influences of ethanol and omega3-fatty acids on: VIII. the posttranslational modifications of apoE at the subcellular level? IX. the mechanism of inhibition of hepatic sialyltransferase activity? Specifically, does ethanol inhibit the synthesis of sialyltransferase and by down regulation of its hepatic mRNA levels?