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具有脂质积累和氧化应激的改良饮食肝脏模型的综合自发荧光特征

Integrated autofluorescence characterization of a modified-diet liver model with accumulation of lipids and oxidative stress.

作者信息

Croce Anna Cleta, Ferrigno Andrea, Piccolini Valeria Maria, Tarantola Eleonora, Boncompagni Eleonora, Bertone Vittorio, Milanesi Gloria, Freitas Isabel, Vairetti Mariapia, Bottiroli Giovanni

机构信息

IGM-CNR, Via Abbiategrasso 207, 27100 Pavia, Italy ; Department of Biology & Biotechnology, University of Pavia, Via Ferrata 9, 27100 Pavia, Italy.

Internal Medicine and Therapeutics, University of Pavia, Via Ferrata 9, 27100 Pavia, Italy.

出版信息

Biomed Res Int. 2014;2014:803491. doi: 10.1155/2014/803491. Epub 2014 Jun 9.

DOI:10.1155/2014/803491
PMID:25006587
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC4070497/
Abstract

Oxidative stress in fatty livers is mainly generated by impaired mitochondrial β-oxidation, inducing tissue damages and disease progression. Under suitable excitation, light liver endogenous fluorophores can give rise to autofluorescence (AF) emission, the properties of which depend on the organ morphofunctional state. In this work, we characterized the AF properties of a rat liver model of lipid accumulation and oxidative stress, induced by a 1-9-week hypercaloric methionine-choline deficient (MCD) diet administration. The AF analysis (excitation at 366 nm) was performed in vivo, via fiber optic probe, or ex vivo. The contribution of endogenous fluorophores involved in redox reactions and in tissue organization was estimated through spectral curve fitting analysis, and AF results were validated by means of different histochemical and biochemical assays (lipids, collagen, vitamin A, ROS, peroxidised proteins, and lipid peroxidation -TBARS-, GSH, and ATP). In comparison with the control, AF spectra changes found already at 1 week of MCD diet reflect alterations both in tissue composition and organization (proteins, lipopigments, and vitamin A) and in oxidoreductive pathway engagement (NAD(P)H, flavins), with a subsequent attempt to recover redox homeostasis. These data confirm the AF analysis potential to provide a comprehensive diagnostic information on negative effects of oxidative metabolism alteration.

摘要

脂肪肝中的氧化应激主要由线粒体β-氧化受损产生，会导致组织损伤和疾病进展。在适当的激发下，肝脏内源性荧光团可产生自发荧光（AF）发射，其特性取决于器官的形态功能状态。在本研究中，我们对由1 - 9周高热量蛋氨酸-胆碱缺乏（MCD）饮食诱导的脂质积累和氧化应激大鼠肝脏模型的AF特性进行了表征。AF分析（366 nm激发）通过光纤探头在体内或离体进行。通过光谱曲线拟合分析估计参与氧化还原反应和组织构建的内源性荧光团的贡献，并通过不同的组织化学和生化检测（脂质、胶原蛋白、维生素A、活性氧、过氧化蛋白质和脂质过氧化 -TBARS-、谷胱甘肽和三磷酸腺苷）验证AF结果。与对照组相比，在MCD饮食1周时就发现的AF光谱变化反映了组织组成和结构（蛋白质、脂色素和维生素A）以及氧化还原途径参与情况（烟酰胺腺嘌呤二核苷酸（磷酸）、黄素）的改变，随后有恢复氧化还原稳态的尝试。这些数据证实了AF分析在提供关于氧化代谢改变负面影响的全面诊断信息方面的潜力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d3b/4070497/a9289dc985a9/BMRI2014-803491.010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d3b/4070497/8b5f2e5f07dd/BMRI2014-803491.001.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d3b/4070497/ceaf684d502b/BMRI2014-803491.007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d3b/4070497/a1697402a574/BMRI2014-803491.008.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d3b/4070497/d5d6261f10a1/BMRI2014-803491.009.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d3b/4070497/a9289dc985a9/BMRI2014-803491.010.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d3b/4070497/8b5f2e5f07dd/BMRI2014-803491.001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d3b/4070497/39c6c1febf0c/BMRI2014-803491.002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d3b/4070497/31ff3572387b/BMRI2014-803491.003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d3b/4070497/8177a8328904/BMRI2014-803491.004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d3b/4070497/d4be1bebb816/BMRI2014-803491.005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d3b/4070497/94f16fa8dd4a/BMRI2014-803491.006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d3b/4070497/ceaf684d502b/BMRI2014-803491.007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d3b/4070497/a1697402a574/BMRI2014-803491.008.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d3b/4070497/a9289dc985a9/BMRI2014-803491.010.jpg

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