The Role and Mechanism of CREBH Regulating SIRT3 in Metabolic Associated Fatty Liver Disease
The Role and Mechanism of CREBH Regulating SIRT3 in Metabolic Associated Fatty Liver Disease
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CREBH调控SIRT3在代谢相关脂肪肝中的作用及机制
DOI：
10.1016/j.lfs.2022.120838
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Xu Keshu
中科院分区：
文献类型：
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作者：
Zhang Junli;Wang Shuhan;Zhao Yajuan;Deng Xiaoling;Li Jiahuan;Xu Keshu
探讨cAMP反应元件结合蛋白H （CREBH）通过调节sirtuin 3 （SIRT3）对代谢性脂肪性肝病的影响。构建蛋氨酸-胆碱缺乏（MCD）和高脂（HF）饮食诱导的脂肪肝小鼠模型，以及棕榈酸（PA）诱导的脂质超载肝细胞体外模型，检测CREBH、SIRT3、总乙酰化、下游蛋白相互作用和脂质代谢表型的表达，并在CREBH - / -小鼠和慢病毒过表达CREBH肝细胞中进一步验证。在脂肪肝和脂质超载模型中，CREBH和SIRT3表达下调，表达呈正相关，同时总蛋白乙酰化水平升高。在体外实验中，CREBH的过表达减轻了肝细胞过量脂质积累、活力受损以及通过脂肪酸氧化途径代谢能量的能力。此外，CREBH的过表达恢复了脱乙酰酶SIRT3与参与脂肪酸氧化途径的肉碱棕榈酰转移酶2 （CPT2）和长链酰基辅酶a脱氢酶（ACADL）分子的相互作用及其脱乙酰状态。然而，CREBH−/−加重了小鼠肝组织脂质代谢的损伤。CREBH通过正向调节SIRT3的表达，提高下游因子的酶活性，促进肝细胞脂肪酸的氧化分解，在MAFLD的脂肪酸氧化中发挥重要作用。缩写：CREBH， cAMP反应元件结合蛋白H；SIRT3, sirtuin 3；CPT2，肉碱棕榈酰转移酶2；ACADL，长链酰基CoA脱氢酶；MCD，蛋氨酸-胆碱缺乏；高频、高脂肪;PA：棕榈酸；MAFLD，代谢性脂肪性肝病；CD，鼠粮；MOI，最优感染多重度；实时定量聚合酶链式反应；K-Ac赖氨酸乙酰化；PPARα，过氧化物酶体增殖物激活受体；OE,超表达;MCAD，中链酰基辅酶a脱氢酶；长链酰基辅酶a脱氢酶；ACAT1，乙酰辅酶a乙酰转移酶1；烯酰辅酶a水合酶1；HMGCS2, 3-羟基-3-甲基戊二酰辅酶a合成酶2；PTMs，翻译后修饰。•CREBH的表达与SIRT3呈正相关。•CREBH通过上调SIRT3减轻肝细胞脂质积累。•CREBH通过上调SIRT3促进参与脂肪酸氧化的CPT2和ACADL的去乙酰化和酶激活。CREBH缺乏会加重MAFLD小鼠的脂质代谢紊乱。
To investigate the effect of cAMP response element-binding protein H (CREBH) on metabolic associated fatty liver disease by regulating sirtuin 3 (SIRT3). Two mouse models of fatty liver induced by a methionine-choline deficient (MCD) diet and a high-fat (HF) diet and an in vitro model of palmitic acid (PA) induced lipid-overloaded hepatocytes were constructed to detect the expression of CREBH, SIRT3, total acetylation, and downstream protein interactions and lipid metabolism phenotype, which were further validated in CREBH −/− mice and lentivirus-overexpressing CREBH hepatocytes. In fatty liver and lipid overload models, the expressions of CREBH and SIRT3 were down-regulated and their expression was positively correlated, accompanied by an increase in the level of total protein acetylation. Overexpression of CREBH alleviated excess lipid accumulation, impaired viability, and the ability to metabolize energy through the fatty acid oxidation pathway in hepatocytes in vitro. Furthermore, overexpression of CREBH restored the interaction of the deacetylase SIRT3 with the molecules carnitine palmitoyl-transferase 2 (CPT2) and long-chain acyl CoA dehydrogenase (ACADL) involved in the fatty acid oxidation pathway and their deacetylation status. However, CREBH −/− aggravated the damage of lipid metabolism in the liver tissue of mice. CREBH increased the enzymatic activity of downstream factors by positively regulating the expression of SIRT3, which promoted the oxidative decomposition of fatty acids in hepatocytes and played an important role in fatty acid oxidation in MAFLD. Abbreviations: CREBH, cAMP response element-binding protein H; SIRT3, sirtuin 3; CPT2, carnitine palmitoyl-transferase 2; ACADL, long-chain acyl CoA dehydrogenase; MCD, methionine-choline deficient; HF, high-fat; PA, palmitic acid; MAFLD, metabolic associated fatty liver disease; CD, chow diet; MOI, optimal multiplicity of infection; qRT-PCR, quantitative real-time polymerase chain reaction; K-Ac, lysine acetylation; PPARα, peroxisome proliferator-activated receptor alpha; OE, overexpression; MCAD, medium-chain acyl-CoA dehydrogenase; ACADVL, very-long-chain acyl-CoA dehydrogenase; ACAT1, acetyl-CoA acetyltransferase 1; ECHS1, enoyl-CoA hydratase-1; HMGCS2, 3-hydroxy-3-methylglutaryl-CoA synthetase 2; PTMs, post-translational modifications. • The expression of CREBH is positively correlated with SIRT3. • CREBH alleviates hepatocyte lipid accumulation by upregulating SIRT3. • CREBH promotes deacetylation and enzyme activation of CPT2 and ACADL involved in fatty acid oxidation by upregulating SIRT3. • CREBH deficiency aggravates lipid metabolism disorders in MAFLD mice.