A Comparison of the Antioxidant Effects Between Hydrogen Gas Inhalation and Vitamin C Supplementation in Response to a 60-Min Treadmill Exercise in Rat Gastrocnemius Muscle

Abstract

The reactive oxygen species (ROS) produced during exercise act as a double-edged sword because they may cause oxidative damage but also play a role in the signaling pathways. A supplementation of exogenous antioxidants can reduce the total amount of ROS during exercise while it may also affect the ROS’ role in the signaling pathways of mitochondrial biogenesis. It has been suggested that hydrogen gas, as an antioxidant, can selectively scavenge hydroxyl radicals but does not affect superoxide anion’s signal transduction. The aim of this study was to compare the effects of 1-h hydrogen gas inhalation 30min prior to a treadmill exercise on the key biomarkers of mitochondrial biogenesis and related signaling pathways, and the activities of endogenous antioxidant enzymes, with those of vitamin C, in the rat skeletal muscle. Eighty-one 8-week-old male Sprague–Dawley (SD) rats were randomly assigned to three interventions (exercise-only, exercise+4%H2, and exercise+vitamin C at 500mg/kg weight, with 27 rats under each intervention), and sampled at pre-, immediately post and 4h post a 60-min treadmill exercise at speed of 27m/min and flat inclination, with nine rats in each sub-group. Expression of mitochondrial biogenetic markers and related signaling molecules in gastrocnemius muscle, and concentrations of oxidative stress markers in serum were measured. Two-way ANOVA or Kruskal–Wallis analyses showed that both hydrogen inhalation and vitamin C supplementation significantly reduced serum levels of MDA immediately after exercise and AGEs 4h after exercise. The pre-exercise supplement of vitamin C significantly reduced mitochondrial complex IV concentrations and PGC-1α, NRF-1, TFAM gene expression levels compared to the pre-exercise group, but the hydrogen gas intervention did not result in a reduction in these measurements. Unlike vitamin C, hydrogen inhalation did not blunt post-exercise mitochondrial biogenetic signals, but resulted in an increase in complex IV concentration, activation of PGC-1α, and TFAM and NRF-2 gene transcription, and up-regulation of PGC-1α protein expression. The findings indicated that hydrogen gas inhalation could play the role as an effective antioxidant in response to the exercise, whilst it did not significantly affect mitochondrial biogenesis. The dose–response relationship and antioxidant effects in different types of exercise for hydrogen inhalation require further investigation.

Mitochondria: one of the vital hubs for molecular hydrogen’s biological functions

“…The promotion of mitochondrial biogenesis by H 2 has been previously reported, as demonstrated by increased expression of mtDNA (mtCoxII, mtCoxIV, and mtNd1) and mitochondrial biogenesis-related genes (Pgc-1α, Tfam, Nrf-1/2, and Pparγ) (Chaoqun et al, 2021;Xie et al, 2021;Luo et al, 2022;Zhang et al, 2023). The H 2 -induced mitochondrial biogenesis may be mediated by PPARγ/AMPK/SIRT1-PGC-1α-NRF1/2-TFAM pathway (Chaoqun et al, 2021;Xie et al, 2021;Luo et al, 2022). In addition to the regulation at transcriptional level, mitochondrial biogenesis can also be governed by post-translational processes, such as mitochondrial protein import regulation (Devaux et al, 2010).…”

“…The activation of this PGC-1α/NRF-1/2/TFAM pathway leads to the synthesis of mitochondrial DNA and proteins and generation of new mitochondria (Popov, 2020). The promotion of mitochondrial biogenesis by H 2 has been previously reported, as demonstrated by increased expression of mtDNA (mtCoxII, mtCoxIV, and mtNd1) and mitochondrial biogenesis-related genes (Pgc-1α, Tfam, Nrf-1/2, and Pparγ) (Chaoqun et al, 2021;Xie et al, 2021;Luo et al, 2022;Zhang et al, 2023). The H 2 -induced mitochondrial biogenesis may be mediated by PPARγ/AMPK/SIRT1-PGC-1α-NRF1/2-TFAM pathway (Chaoqun et al, 2021;Xie et al, 2021;Luo et al, 2022).…”

Therapeutic Potential of Hydrogen-Rich Water on Muscle Atrophy Caused by Immobilization in a Mouse Model

“…Animal studies demonstrate that H 2 treatment attenuated the exercise-induced increases in skeletal muscle TNF-α, NF-κB, IL-6, and caspase-3 levels while increasing superoxide dismutase levels [20]. Moreover, another study found that compared to but unlike vitamin C, H 2 treatment did not reduce the exercise-induced elevated levels of peroxisome proliferator-activated receptor-gamma (PPARγ) coactivator-1alpha (PGC-1α), nuclear transcription factors 1-2 (NRF-1,2), and mitochondrial transcription factor A (TFAM) [27]. These studies indicate that H 2 can reduce excessive levels of inflammation and oxidative stress, but does not interfere with favorable exercise-induced adaptations in the skeletal muscle.…”

The effects of advanced glycation end‐products on skin and potential anti‐glycation strategies

“…The antioxidant, such as N-acetylcysteine (NAC), Vitamin C, Vitamin E, can scavenge ROS, prevent oxidative stress and damage. 69,81,82 These are all important approaches to inhibit AGEs.…”

“…Antioxidants can help reduce oxidative stress, lowering AGEs generation. Antioxidants assist in scavenging free radicals, decreasing the oxidation of proteins, lipids and nucleic acids, thereby slowing down AGEs formation 11,35,69 …”