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营养性和遗传性脂肪肝模型中代谢指纹的自体荧光鉴别

Autofluorescence discrimination of metabolic fingerprint in nutritional and genetic fatty liver models.

作者信息

Croce Anna C, Ferrigno Andrea, Di Pasqua Laura G, Berardo Clarissa, Piccolini Valeria Maria, Bertone Vittorio, Bottiroli Giovanni, Vairetti Mariapia

机构信息

Italian National Research Council (CNR), Institute of Molecular Genetics, Via Abbiategrasso 207, Pavia, Italy; University of Pavia, Department of Biology & Biotechnology, Via Ferrata 9, Pavia, Italy.

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

出版信息

J Photochem Photobiol B. 2016 Nov;164:13-20. doi: 10.1016/j.jphotobiol.2016.09.015. Epub 2016 Sep 12.

Abstract

Liver tissue autofluorescence (AF) has been characterized in two models with a different potential to undergo disease progression to steatohepatitis: Wistar rats, administered with a methionine, choline deficient diet (MCD), and Zucker (fa/fa) rats, homozygous for a spontaneous mutation of leptin receptor. AF spectra were recorded from liver tissue cryostatic sections by microspectrofluorometry, under 366nm excitation. Curve fitting analysis was used to estimate the contribution of different endogenous fluorophores (EFs) to the overall AF emission: i) fluorescing fatty acids, a fraction of liver lipids up to now poorly considered and complicated to detect by conventional procedures; ii) lipofuscin-like lipopigments, biomarkers of oxidizing events; iii) NAD(P)H and flavins, biomarkers of energy metabolism and tissue redox state. AF data and biochemical correlates of hepatocellular injury resulted to depend more on rat strain than on intratissue bulk lipid or ROS levels, reflecting a different metabolic ability of the two models to counteract potentially harmful agents. AF analysis can thus be proposed for extensive applications ranging from experimental hepatology to the clinics. AF based diagnostic procedures are expected to help both the prediction of the risk of fatty liver disease progression and the prescreening of marginal organs to be recruited as donors for transplantation. A support is also foreseen in the advancement and personalization of strategies to ameliorate the donor organ preservation outcome and the follow up of therapeutic interventions.

摘要

在两种具有不同疾病进展为脂肪性肝炎可能性的模型中,对肝脏组织自体荧光(AF)进行了表征:给予蛋氨酸、胆碱缺乏饮食(MCD)的Wistar大鼠,以及瘦素受体自发突变的纯合子Zucker(fa/fa)大鼠。通过显微分光荧光测定法,在366nm激发下,从肝脏组织低温切片记录AF光谱。曲线拟合分析用于估计不同内源性荧光团(EFs)对总体AF发射的贡献:i)荧光脂肪酸,这是肝脏脂质的一部分,迄今为止在传统方法中考虑不足且检测复杂;ii)脂褐素样脂色素,氧化事件的生物标志物;iii)NAD(P)H和黄素,能量代谢和组织氧化还原状态的生物标志物。AF数据和肝细胞损伤的生化相关性结果表明,其更多地取决于大鼠品系,而非组织内总脂质或ROS水平,这反映了两种模型在对抗潜在有害物质方面的不同代谢能力。因此,可以提出将AF分析广泛应用于从实验性肝脏病学到临床的各个领域。基于AF的诊断程序有望有助于预测脂肪性肝病进展的风险,并对作为移植供体的边缘器官进行预筛选。在改善供体器官保存结果和治疗干预随访的策略的推进和个性化方面,也可以预见会得到支持。

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