Effect of aging in functional redox state of single isolated skeletal muscle fibres

Abstract

[EN] Skeletal muscle constantly produces reactive oxygen species (ROS). During contractile activity ROS are generated in skeletal muscle fibres. There is considerable support for an involvement of ROS in the process of aging. Several studies indicate that adaptive responses of skeletal muscle that are activated and regulated by ROS are disrupted during aging. The aim of this study was to monitor, in real time, intracellular ROS production in single skeletal muscle fibres from old and young mice and study the effect of contractile activity in these cells. Following evaluate and correlate the potential changes in intracellular ROS production with glutathione redox state and antioxidant enzymatic activities in muscle. Single skeletal muscle fibres were isolated from the Flexor Digitorus Brevis muscle from young (2-4 monthold) and old (26-28 month-old) C57BL/6 mice. Fibres were loaded with DCFH-DA, a fluorophore probe that allows the quantification of intracellular ROS generation by fluorescence microscopy imaging. Contractile activity was induced in fibres by electrical stimulation. Glutathione redox state and activity of antioxidant enzymes were analysed in gastrocnemious muscle. Intracellular basal level of ROS was higher in fibres from old mice. Contractile activity induced increase of ROS generation in fibres from young mice. However, this response was attenuated in fibres from old mice. Glutathione redox state was significant different, in favour of oxidized glutathione, in muscles from old mice. Glutathione peroxidase and catalase activities were significantly augmented in muscles from old mice. In conclusion, the process of aging modifies the basal redox status in skeletal muscle fibres in favour of oxidation and induces adaptation mechanisms of antioxidant defences. These are not able to neutralize the increase of basal oxidation, but they might lead to the attenuation of ROS produced by contractile activity observed in fibres from old mice.