NOVEL MODULATORS OF ALCOHOL INDUCED METABOLIC AND LIVER INJURY

描述（由申请人提供）：理由：慢性乙醇诱导沉默调节基因1 （SIRT1）和过氧化物酶体增殖体受体共激活因子1 (PGC1) 1的下调和PGC12的上调影响肝脏脂质氧化和脂肪生成，导致脂肪肝和随之而来的损伤。我们显示了饮食低？3-脂肪酸？3FA)，但不高？大豆蛋白（SP）具有相似的降脂效果。因此，证明这两种新型调节剂可能通过调节这些对立的PGC11和PGC12信号通路基因来减轻酒精诱导的脂肪肝和损伤具有临床意义。初步研究：我们的基本原理是基于我们的出版记录（过去5年的12篇出版物，1篇综述文章，1本书和21篇摘要），以及总结如下的强有力的初步数据：3FA和SP通过(a)降低血脂并降低脂肪肝来对抗慢性酒精的作用；(b)减弱肝脏PGC12、SREBP1c及其靶脂质途径基因mrna的上调；(c)防止肝脏SIRT1、PGC11及其靶脂肪酸氧化途径基因mrna下调；(d)防止乙酰- pgc11（其无活性形式）在肝脏中的增加；(e)恢复受损的肝脂肪酸氧化和糖异生；(f)恢复肝脏中因慢性酒精治疗而降低的PO4AMPK水平。(g)通过Western blots，我们证实了慢性酒精引起的PGC11和PGC12信号通路基因mrna及相应酶蛋白水平的变化。(h) PGC11在酒精性脂肪肝中起关键作用，PGC11- ko小鼠肝脂质评分显著升高。(i) PGC12似乎起着次要作用，因为PGC12- ko小鼠仍然发展为酒精性脂肪肝，尽管不像PGC11- ko小鼠那么明显，可能是因为乙醇介导的PGC11的强烈抑制。具体目标：根据这些基本原理和初步调查结果，我们将解决以下具体目标：的影响？3FA和SP对慢性乙醇诱导的血浆和肝脏脂质、脂蛋白、AST、ALT升高的影响及其与肝脏组织病理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、脂质生成基因和新生脂质合成率上调的影响。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下调的影响。方法：PI和他的团队将利用他们在分子生物学，免疫和组织化学以及生化方法方面的专业知识，以啮齿动物为实验模型来实现这些特定目标。PI还将使用PGC11-KO和PGC12-KOs小鼠模型来最终证明它们在乙醇、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.