NOVEL MODULATORS OF ALCOHOL INDUCED METABOLIC AND LIVER INJURY

描述（由申请方提供）：依据：慢性乙醇诱导的沉默调节基因1（SIRT 1）和过氧化物酶体增殖物受体辅激活因子1（PGC1）1下调以及PGC12上调影响肝脏脂质氧化和脂肪生成，导致脂肪肝和随后的损伤。我们的饮食低？3-脂肪酸（？3FA），但不高？3FA显著降低血清和肝脏脂质，而大豆蛋白（SP）具有类似的降脂作用。因此，证明这两种新型调节剂通过调节这些相反的PGC11和PGC12信号通路基因而潜在地减轻酒精诱导的脂肪肝和损伤是临床相关的。初步研究：我们的理由是基于我们的出版记录（12篇出版物，1篇评论文章，1本书和21篇摘要在过去5年），并在强大的初步数据，总结如下：低？3FA和SP通过（a）降低血脂并减少脂肪肝，（B）减弱肝脏PGC12、SREBP 1c及其靶脂肪生成途径基因mRNA的上调，（c）阻止肝脏SIRT 1和PGC11及其靶脂肪酸氧化途径基因mRNA的下调，（d）抑制肝脏脂肪酸氧化途径基因mRNA的上调，（e）抑制肝脏脂肪酸氧化途径基因mRNA的下调，（f）抑制肝脏脂肪酸氧化途径基因mRNA的上调，（f）抑制肝脏脂肪酸氧化途径基因mRNA的下调，（f）抑制肝脏脂肪酸氧化途径基因mRNA的上调，（f）抑制肝脏脂肪酸氧化途径基因mRNA的下调。（d）防止乙酰-PGC11（其失活形式）的肝脏增加;（e）恢复受损的肝脏脂肪酸氧化和再生;和（f）恢复因长期酒精治疗而降低的PO 4 AMPK的肝脏水平。(g)通过蛋白质印迹，我们证明了慢性酒精引起的PGC 11和PGC 12信号通路基因mRNA水平以及相应酶蛋白的伴随变化。(h)PGC11在酒精诱导的脂肪肝中起关键作用，这通过PGC11-KO小鼠中肝脏脂质评分的显著增加来证明。(i)PGC12似乎起次要作用，因为PGC12-KO小鼠仍发生酒精性脂肪肝，尽管不如PGC11-KO小鼠明显，可能是因为乙醇介导的PGC11强烈抑制。具体目标：基于这些基本原理和初步调查结果，我们将讨论以下具体目标：具体目标1。影响？3FA和SP对慢性乙醇所致血脂、肝脂蛋白、AST、ALT升高的影响及其与肝组织学的相关性具体目标2。的作用机制？3FA和SP对乙醇诱导的肝脏脂质合成代谢转录辅激活因子、脂肪生成基因和脂质合成速率上调的影响。具体目标3。的作用机制？3FA和SP对乙醇诱导的肝脏脂质分解代谢转录辅激活因子、脂质氧化基因和脂肪酸氧化速率的下调。接近方法：PI和他的团队将通过使用啮齿动物作为实验模型，利用他们在分子生物学，免疫和组织化学以及生物化学方法方面的专业知识来实现这些特定目标。PI还将使用PGC11-KO和PGC12-科斯小鼠模型来最终证明其在乙醇、低3FA和SP通过这些信号通路引起/预防酒精性脂肪肝和损伤的作用中的特异性。

DESCRIPTION (provided by applicant): Rationale: Chronic ethanol-induced down regulations of silence regulator gene 1 (SIRT1) and peroxisome proliferator receptor coactivator 1 (PGC1) 1 and up-regulation of PGC12 affect hepatic lipid oxidation and lipogenesis leading to fatty liver and consequent injury. We show that dietary low ?3-fatty acids (?3FA), but not high ?3FA, significantly decreased serum and liver lipids, while Soy Proteins (SP) had similar lipid lowering effects. Therefore, it is clinically relevant to prove that these two novel modulators attenuate alcohol-induced fatty liver and injury potentially by regulating these opposing PGC11 and PGC12 signaling pathway genes. Preliminary Studies: Our rationales are based on our publication records (12 Publications, 1 Review article, 1 book & 21 Abstracts in the past 5 years), and on strong preliminary data that are summarized below: Low ?3FA & SP opposed the actions of chronic alcohol by (a) Reducing serum lipids with concomitant decreased fatty liver; (b) Attenuating the up-regulation of hepatic mRNAs of PGC12, SREBP1c and their target lipogenic pathway genes; (c) Preventing the down-regulation of hepatic mRNAs of SIRT1 and PGC11 and their target fatty acid oxidation pathway genes; (d) Preventing the hepatic increase in acetyl-PGC11, its inactive form; (e) Restoring impaired hepatic fatty acid oxidation and gluconeogenesis; And (f) Restoring the hepatic level of PO4AMPK that was decreased by chronic alcohol treatment. (g) By Western blots, we demonstrated concomitant changes in levels of mRNAs of PGC11 and PGC12 signaling pathway genes and the corresponding enzyme proteins caused by chronic alcohol. (h) That PGC11 plays a critical role in alcohol- induced fatty liver was demonstrated by dramatic increase in hepatic lipid score in PGC11-KO mice. (i) PGC12 seems to play a secondary role because PGC12-KO mice still developed alcoholic fatty liver albeit less pronounced than in PGC11-KO mice, presumably because of ethanol-mediated strong inhibition of PGC11. Specific aims: Based on these rationales and preliminary findings we will address the following specific aims: Specific Aim 1. Effects of ?3FA and SP on Chronic Ethanol-induced Increases in Plasma and Hepatic Lipids and Lipoproteins, AST, ALT and Their Correlation with Liver Histopathology. Specific Aim 2. Mechanisms of Actions of ?3FA and SP on Ethanol-induced Up-regulation of Hepatic Lipid Anabolic Transcriptional Coactivators, Lipogenic Genes and de novo Lipid Synthetic Rates. Specific Aim 3. Mechanisms of Actions of ?3FA & SP on Ethanol-induced Down-regulation of Hepatic Lipid Catabolic Transcriptional Coactivators, Lipid Oxidizing Genes & Fatty Acid Oxidation Rates. Methods of Approach: PI and his team will accomplish these specific aims by using rodents as experimental models with their expertise in molecular biology, immuno- and histo-chemistry and biochemical approaches. PI will also use PGC11-KO and PGC12-KOs mouse models to conclusively prove their specificities in the actions of ethanol, Low 3FA & SP via these signaling pathways in causing/preventing alcoholic fatty liver and injury.