Blottner Dieter, Capitanio Daniele, Trautmann Gabor, Furlan Sandra, Gambara Guido, Moriggi Manuela, Block Katharina, Barbacini Pietro, Torretta Enrica, Py Guillaume, Chopard Angèle, Vida Imre, Volpe Pompeo, Gelfi Cecilia, Salanova Michele
Institute of Integrative Neuroanatomy, Charité-Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 10115 Berlin, Germany.
Center of Space Medicine Berlin, 10115 Berlin, Germany.
Antioxidants (Basel). 2021 Mar 3;10(3):378. doi: 10.3390/antiox10030378.
Increased oxidative stress by reactive oxygen species (ROS) and reactive nitrogen species (RNS) is a major determinant of disuse-induced muscle atrophy. Muscle biopsies (thigh vastus lateralis, ) obtained from healthy male subjects enrolled in the Toulouse Cocktail bedrest (BR) study were used to assess efficacy of an antioxidant cocktail (polyphenols, omega-3, vitamin E, and selenium) to counteract the increased redox homeostasis and enhance the antioxidant defense response by using label-free LC-MS/MS and NITRO-DIGE (nitrosated proteins), qPCR, and laser confocal microscopy. Label-free LC-MS/MS indicated that treatment prevented the redox homeostasis dysregulation and promoted structural remodeling (TPM3, MYH7, MYBPC, MYH1, MYL1, HRC, and LUM), increment of RyR1, myogenesis (CSRP3), and skeletal muscle development (MUSTN1, LMNA, AHNAK). These changes were absent in the Placebo group. Glycolysis, tricarboxylic acid cycle (TCA), oxidative phosphorylation, fatty acid beta-oxidation, and mitochondrial transmembrane transport were normalized in treated subjects. Proteins involved in protein folding were also normalized, whereas protein entailed in ion homeostasis decreased. NITRO-DIGE analysis showed significant protein nitrosylation changes for CAT, CA3, SDHA, and VDAC2 in Treatment vs. Placebo. Similarly, the nuclear factor erythroid 2-related factor 2 (Nrf-2) antioxidant response element (Nrf-2 ARE) signaling pathway showed an enhanced response in the Treatment group. Increased nitrosative redox homeostasis and decreased antioxidant defense response were found in post-BR control (Placebo, = 10) vs. the antioxidant cocktail treated group (Treatment, = 10). Taken together, increased nitrosative redox homeostasis and muscle deterioration during BR-driven physical inactivity were prevented, whereas decreased antioxidant nitrosative stress defense response was attenuated by Treatment suggesting positive effects of the nutritional intervention protocol in bedrest.
活性氧（ROS）和活性氮（RNS）导致的氧化应激增加是废用性肌肉萎缩的主要决定因素。从参与图卢兹鸡尾酒卧床休息（BR）研究的健康男性受试者身上获取的肌肉活检样本（大腿外侧股四头肌），用于评估一种抗氧化鸡尾酒（多酚、ω-3、维生素E和硒）通过无标记液相色谱-串联质谱法（LC-MS/MS）和硝基荧光差异凝胶电泳（NITRO-DIGE，用于检测亚硝基化蛋白质）、定量聚合酶链反应（qPCR）以及激光共聚焦显微镜，来对抗氧化还原稳态增加并增强抗氧化防御反应的功效。无标记LC-MS/MS表明，该治疗可防止氧化还原稳态失调，并促进结构重塑（肌动蛋白3、肌球蛋白重链7、肌球蛋白结合蛋白C、肌球蛋白重链1、肌球蛋白轻链1、肌质网钙释放通道、层黏连蛋白）、兰尼碱受体1（RyR1）增加、成肌作用（富含半胱氨酸的蛋白3）以及骨骼肌发育（肌肉相关转录因子1、核纤层蛋白A/C、巨蛋白）。安慰剂组未出现这些变化。在接受治疗的受试者中，糖酵解、三羧酸循环（TCA）、氧化磷酸化、脂肪酸β氧化以及线粒体跨膜转运均恢复正常。参与蛋白质折叠的蛋白质也恢复正常，而参与离子稳态的蛋白质减少。NITRO-DIGE分析显示，与安慰剂组相比，治疗组中过氧化氢酶（CAT）、碳酸酐酶3（CA3）、琥珀酸脱氢酶A（SDHA）和电压依赖性阴离子通道2（VDAC2）的蛋白质亚硝基化有显著变化。同样，核因子红细胞2相关因子2（Nrf-2）抗氧化反应元件（Nrf-2 ARE）信号通路在治疗组中显示出增强的反应。与抗氧化鸡尾酒治疗组（治疗组，n = 10）相比，BR后对照组（安慰剂组，n = 10）的亚硝化氧化还原稳态增加，抗氧化防御反应降低。综上所述，BR导致的身体不活动期间亚硝化氧化还原稳态增加和肌肉退化得到了预防，而治疗减轻了抗氧化亚硝化应激防御反应的降低，表明营养干预方案对卧床休息有积极作用。
