The Impact of Spermidine on C2C12 Myoblasts Proliferation, Redox Status and Polyamines Metabolism under H2O2 Exposure

Abstract

A central feature of the skeletal muscle is its ability to regenerate through the activation, by environmental signals, of satellite cells. Once activated, these cells proliferate as myoblasts, and defects in this process profoundly affect the subsequent process of regeneration. High levels of reactive oxygen species such as hydrogen peroxide (H2O2) with the consequent formation of oxidized macromolecules increase myoblasts’ cell death and strongly contribute to the loss of myoblast function. Recently, particular interest has turned towards the beneficial effects on muscle of the naturally occurring polyamine spermidine (Spd). In this work, we tested the hypothesis that Spd, upon oxidative challenge, would restore the compromised myoblasts’ viability and redox status. The effects of Spd in combination with aminoguanidine (Spd-AG), an inhibitor of bovine serum amine oxidase, on murine C2C12 myoblasts treated with a mild dose of H2O2 were evaluated by analyzing: (i) myoblast viability and recovery from wound scratch; (ii) redox status and (iii) polyamine (PAs) metabolism. The treatment of C2C12 myoblasts with Spd-AG increased cell number and accelerated scratch wound closure, while H2O2 exposure caused redox status imbalance and cell death. The combined treatment with Spd-AG showed an antioxidant effect on C2C12 myoblasts, partially restoring cellular total antioxidant capacity, reducing the oxidized glutathione (GSH/GSSG) ratio and increasing cell viability through a reduction in cell death. Moreover, Spd-AG administration counteracted the induction of polyamine catabolic genes and PA content decreased due to H2O2 challenges. In conclusion, our data suggest that Spd treatment has a protective role in skeletal muscle cells by restoring redox balance and promoting recovery from wound scratches, thus making myoblasts able to better cope with an oxidative insult.

Morroniside Protects C2C12 Myoblasts from Oxidative Damage Caused by ROS-Mediated Mitochondrial Damage and Induction of Endoplasmic Reticulum Stress

“…It stands to reason therefore that increasing GSH levels may be an effective strategy to protect cells from oxidative damage. Similar to results seen previously ( Ceci et al ., 2022a , 2022b ), we show that reductions to GSH levels in H 2 O 2 -treated C2C12 cells can be significantly blocked by morroniside pretreatment. Analysis of cellular ROS levels using DCF-DA showed that intracellular ROS production was significantly increased by H 2 O 2 stimulation, and that these were clearly restored to more normal levels through co-treatment of H 2 O 2 with morroniside.…”

Spermidine improves the antioxidant capacity and morphology of intestinal tissues and regulates intestinal microorganisms in Sichuan white geese

“…Spermine supplementation during lactation can improve the intestinal villus height: crypt depth ratio and intestinal absorption area of piglets, thus promoting their growth ( van Wettere et al, 2016 ). Spermidine can improve the antioxidant capacity of muscle cells and reduce the ratio of oxidized glutathione (GSH/GSSH) to promote cell proliferation ( Ceci et al, 2022 ). In addition, spermidine was able to reverse the decreased activity of antioxidant enzymes in the hearts of offspring caused by hypoxia in utero ( Chai et al, 2022 ).…”

Embryonic Cadaverine Signaling: Implications for Plasma Free Amino Acid and Skeletal Muscle Energy Metabolism in Newly Hatched Chicks

“…Polyamines are generic terms for cationic organic compounds containing multiple amino groups. The chemical characteristics of polyamines allow them to function in cellular homeostasis, including in cell proliferation and differentiation[ 2 ], nucleic acid protection[ 3 ], apoptosis[ 4 ], autophagy[ 5 ], and antioxidant activity[ 6 ]. Other common polyamines found in vertebrates include putrescine, spermidine, and spermine, which are synthesized by the decarboxylation of ornithine[ 7 ].…”