Gonçalves Inês O, Maciel Elisabete, Passos Emanuel, Torrella Joan R, Rizo David, Viscor Ginés, Rocha-Rodrigues Silvia, Santos-Alves Estela, Domingues Maria R, Oliveira Paulo J, Ascensão António, Magalhães José
Research Center in Physical Activity, Health and Leisure, Faculty of Sport, University of Porto, Portugal.
Mass Spectrometry Centre, Chemistry Department, University of Aveiro, Portugal.
Int J Biochem Cell Biol. 2014 Sep;54:163-73. doi: 10.1016/j.biocel.2014.07.011. Epub 2014 Jul 23.
Mitochondrial membrane lipid composition is a critical factor in non-alcoholic steatohepatitis (NASH). Exercise is the most prescribed therapeutic strategy against NASH and a potential modulator of lipid membrane. Thus, we aimed to analyze whether physical exercise exerted preventive (voluntary physical activity - VPA) and therapeutic (endurance training - ET) effect on NASH-induced mitochondrial membrane changes. Sprague-Dawley rats (n=36) were divided into standard-diet sedentary (SS, n=12), standard-diet VPA (SVPA, n=6), high-fat diet sedentary (HS, n=12) and high-fat diet VPA (HVPA, n=6). After 9 weeks of diet-specific feeding, half of SS and HS group were engaged in an ET program for 8 weeks/5 day/week/1h/day (SET, HET). Liver mitochondria were isolated for oxygen consumption and transmembrane-electric potential (ΔΨ) assays. Mitochondrial phospholipid classes and fatty acids were quantified through thin layer chromatography and gas chromatography, respectively, while cardiolipin (CL), phosphatidylcholine (PC) phosphatidylethanolamine (PE) and phosphatidylinositol (PI) molecular profile was determined by electrospray mass spectrometry. In parallel with histological signs of NASH, high-fat diet decreased PI, CL and PC/PE ratio, whereas PE and phosphatidic acid content increased in sedentary animals (HS vs. SS). Moreover, a decrease in linolelaidic, monounsaturated fatty acids content and an increase in saturated fatty acids (SFAS) were observed. Along with phospholipidomic alterations, HS animals showed a decrease in respiratory control ratio (RCR), ΔΨ and FCCP-induced uncoupling respiration (HS vs. SS). Both phospholipidomic (PC/PE, SFAS) and mitochondrial respiratory alterations were counteracted by exercise interventions. Exercise used as preventive (VPA) or therapeutic (ET) strategies preserved liver mitochondrial phospholipidomic profile and maintained mitochondrial function in a model of NASH.
线粒体膜脂质组成是非酒精性脂肪性肝炎（NASH）的一个关键因素。运动是针对NASH最常用的治疗策略，也是脂质膜的潜在调节因子。因此，我们旨在分析体育锻炼是否对NASH诱导的线粒体膜变化产生预防（自愿体育活动 - VPA）和治疗（耐力训练 - ET）作用。将36只Sprague-Dawley大鼠分为标准饮食久坐组（SS，n = 12）、标准饮食VPA组（SVPA，n = 6）、高脂饮食久坐组（HS，n = 12）和高脂饮食VPA组（HVPA，n = 6）。在进行9周特定饮食喂养后，SS组和HS组的一半大鼠参与了为期8周、每周5天、每天1小时的ET计划（SET，HET）。分离肝脏线粒体用于耗氧量和跨膜电位（ΔΨ）测定。分别通过薄层色谱法和气相色谱法对线粒体磷脂类别和脂肪酸进行定量，同时通过电喷雾质谱法测定心磷脂（CL）、磷脂酰胆碱（PC）、磷脂酰乙醇胺（PE）和磷脂酰肌醇（PI）的分子谱。与NASH的组织学特征一致，高脂饮食使久坐动物（HS组与SS组相比）的PI、CL和PC/PE比值降低，而PE和磷脂酸含量增加。此外，还观察到亚油酸、单不饱和脂肪酸含量降低，饱和脂肪酸（SFA）含量增加。除了磷脂组学改变外，HS组动物的呼吸控制率（RCR）、ΔΨ和FCCP诱导的解偶联呼吸均降低（HS组与SS组相比）。运动干预抵消了磷脂组学（PC/PE，SFA）和线粒体呼吸的改变。在NASH模型中，将运动用作预防（VPA）或治疗（ET）策略可保留肝脏线粒体磷脂组学特征并维持线粒体功能。
