Proteomic identification and characterization of hepatic glyoxalase 1 dysregulation in non-alcoholic fatty liver disease.

IF 2.1 3区生物学 Q3 BIOCHEMICAL RESEARCH METHODS

Proteome Science Pub Date : 2018-02-14 eCollection Date: 2018-01-01 DOI:10.1186/s12953-018-0131-y

Christos Spanos, Elaina M Maldonado, Ciarán P Fisher, Petchpailin Leenutaphong, Ernesto Oviedo-Orta, David Windridge, Francisco J Salguero, Alexandra Bermúdez-Fajardo, Mark E Weeks, Caroline Evans, Bernard M Corfe, Naila Rabbani, Paul J Thornalley, Michael H Miller, Huan Wang, John F Dillon, Alberto Quaglia, Anil Dhawan, Emer Fitzpatrick, J Bernadette Moore

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引用次数: 20

Abstract

Background: Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease worldwide. However, its molecular pathogenesis is incompletely characterized and clinical biomarkers remain scarce. The aims of these experiments were to identify and characterize liver protein alterations in an animal model of early, diet-related, liver injury and to assess novel candidate biomarkers in NAFLD patients.

Methods: Liver membrane and cytosolic protein fractions from high fat fed apolipoprotein E knockout (ApoE^-/-) animals were analyzed by quantitative proteomics, utilizing isobaric tags for relative and absolute quantitation (iTRAQ) combined with nano-liquid chromatography and tandem mass spectrometry (nLC-MS/MS). Differential protein expression was confirmed independently by immunoblotting and immunohistochemistry in both murine tissue and biopsies from paediatric NAFLD patients. Candidate biomarkers were analyzed by enzyme-linked immunosorbent assay in serum from adult NAFLD patients.

Results: Through proteomic profiling, we identified decreased expression of hepatic glyoxalase 1 (GLO1) in a murine model. GLO1 protein expression was also found altered in tissue biopsies from paediatric NAFLD patients. In vitro experiments demonstrated that, in response to lipid loading in hepatocytes, GLO1 is first hyperacetylated then ubiquitinated and degraded, leading to an increase in reactive methylglyoxal. In a cohort of 59 biopsy-confirmed adult NAFLD patients, increased serum levels of the primary methylglyoxal-derived advanced glycation endproduct, hydroimidazolone (MG-H1) were significantly correlated with body mass index (r = 0.520, p < 0.0001).

Conclusion: Collectively these results demonstrate the dysregulation of GLO1 in NAFLD and implicate the acetylation-ubquitination degradation pathway as the functional mechanism. Further investigation of the role of GLO1 in the molecular pathogenesis of NAFLD is warranted.

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非酒精性脂肪肝中肝乙草醛酶1失调的蛋白质组学鉴定和特征

背景:非酒精性脂肪性肝病(NAFLD)是世界范围内最常见的肝脏疾病。然而，其分子发病机制尚不完全清楚，临床生物标志物仍然缺乏。这些实验的目的是鉴定和表征早期饮食相关肝损伤动物模型中的肝蛋白改变，并评估NAFLD患者新的候选生物标志物。方法:采用定量蛋白质组学方法，结合纳米液相色谱和串联质谱(nLC-MS/MS)技术，对高脂喂养载脂蛋白E基因敲除(ApoE-/-)动物的肝膜和细胞质蛋白进行相对定量和绝对定量等压标记(iTRAQ)分析。通过免疫印迹和免疫组织化学分别在小鼠组织和小儿NAFLD患者的活检中证实了差异蛋白的表达。采用酶联免疫吸附法分析成人NAFLD患者血清中的候选生物标志物。结果:通过蛋白质组学分析，我们在小鼠模型中发现肝乙二醛酶1 (GLO1)的表达降低。在小儿NAFLD患者的组织活检中也发现GLO1蛋白表达改变。体外实验表明，在肝细胞脂质负荷下，GLO1首先被高乙酰化，然后被泛素化和降解，导致反应性甲基乙二醛增加。在一组59例活检证实的成人NAFLD患者中，原发性甲基乙二醛衍生的晚期糖基化终产物氢咪唑酮(MG-H1)的血清水平升高与体重指数显著相关(r = 0.520, p)。结论:这些结果共同表明了NAFLD中GLO1的失调，并暗示乙酰化-羧化降解途径是其功能机制。GLO1在NAFLD分子发病机制中的作用有待进一步研究。

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来源期刊

Proteome Science 生物-生化研究方法

CiteScore

2.90

自引率

0.00%

发文量

审稿时长

4.5 months

期刊介绍： Proteome Science is an open access journal publishing research in the area of systems studies. Proteome Science considers manuscripts based on all aspects of functional and structural proteomics, genomics, metabolomics, systems analysis and metabiome analysis. It encourages the submissions of studies that use large-scale or systems analysis of biomolecules in a cellular, organismal and/or environmental context. Studies that describe novel biological or clinical insights as well as methods-focused studies that describe novel methods for the large-scale study of any and all biomolecules in cells and tissues, such as mass spectrometry, protein and nucleic acid microarrays, genomics, next-generation sequencing and computational algorithms and methods are all within the scope of Proteome Science, as are electron topography, structural methods, proteogenomics, chemical proteomics, stem cell proteomics, organelle proteomics, plant and microbial proteomics. In spite of its name, Proteome Science considers all aspects of large-scale and systems studies because ultimately any mechanism that results in genomic and metabolomic changes will affect or be affected by the proteome. To reflect this intrinsic relationship of biological systems, Proteome Science will consider all such articles.