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The impact of resveratrol and hydrogen peroxide on muscle cell plasticity shows a dose-dependent interaction.

Bosutti A, Degens H - Sci Rep (2015)

Bottom Line: RS did not increase oxidative capacity.In conclusion, low resveratrol doses promoted in vitro muscle regeneration and attenuated the impact of ROS, while high doses augmented the reduced plasticity and metabolism induced by oxidative stress.Thus, the effects of resveratrol depend on its dose and degree of oxidative stress.

View Article: PubMed Central - PubMed

Affiliation: School of Healthcare Science, Manchester Metropolitan University, Manchester, United Kingdom.

ABSTRACT
While reactive oxygen species (ROS) play a role in muscle repair, excessive amounts of ROS for extended periods may lead to oxidative stress. Antioxidants, as resveratrol (RS), may reduce oxidative stress, restore mitochondrial function and promote myogenesis and hypertrophy. However, RS dose-effectiveness for muscle plasticity is unclear. Therefore, we investigated RS dose-response on C2C12 myoblast and myotube plasticity 1. in the presence and 2. absence of different degrees of oxidative stress. Low RS concentration (10 μM) stimulated myoblast cell cycle arrest, migration and sprouting, which were inhibited by higher doses (40-60 μM). RS did not increase oxidative capacity. In contrast, RS induced mitochondria loss, reduced cell viability and ROS production, and activated stress response pathways [Hsp70 and pSer36-p66(ShcA) proteins]. However, the deleterious effects of H2O2 (1000 µM) on cell migration were alleviated after preconditioning with 10 µM-RS. This dose also enhanced cell motility mediated by 100 µM-H2O2, while higher RS-doses augmented the H2O2-induced impaired myoblast regeneration and mitochondrial dehydrogenase activity. In conclusion, low resveratrol doses promoted in vitro muscle regeneration and attenuated the impact of ROS, while high doses augmented the reduced plasticity and metabolism induced by oxidative stress. Thus, the effects of resveratrol depend on its dose and degree of oxidative stress.

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Resveratrol induced p66Shc-Ser36 phosphorylation and up-regulation of Heat Shock Protein (Hsp)-70.(a) Western blot analysis showing the effect of resveratrol (RS; 10, 20, 40 and 60 µM), H2O2 (100 and 1000 µM) and RS pre-conditioning (10 and 20 µM) on p66Shc-Ser36 phosphorylation and Hsp-70 protein levels. Dose dependent increases in Ser36-p66 phosphorylation and in Hsp-70 protein contents were observed in resveratrol-treated cells. 1000 µM, but not 100 µM H2O2 induced p66Shc(A)-Ser36 phosphorylation (ai); both concentrations of H2O2 elevated Hsp-70 protein levels, but more so in 1000 than 100 µM H2O2 (aii). RS (10 and 20 µM) pre-conditioning attenuated the effects of 1000 µM H2O2 on Hsp-70 (aii), but not on p66Shc-Ser36 phosphorylation (ai). DM vs. CT: in none of the cases was significant. P-values calculated using a two-tailed Student's t-test.*: P<0.01 vs CT; ¤: P<0.001 vs H2O2 1000 µM; ‡: P<0.01 vs H2O2 100 µM. Data are mean ± s.e.m.
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f8: Resveratrol induced p66Shc-Ser36 phosphorylation and up-regulation of Heat Shock Protein (Hsp)-70.(a) Western blot analysis showing the effect of resveratrol (RS; 10, 20, 40 and 60 µM), H2O2 (100 and 1000 µM) and RS pre-conditioning (10 and 20 µM) on p66Shc-Ser36 phosphorylation and Hsp-70 protein levels. Dose dependent increases in Ser36-p66 phosphorylation and in Hsp-70 protein contents were observed in resveratrol-treated cells. 1000 µM, but not 100 µM H2O2 induced p66Shc(A)-Ser36 phosphorylation (ai); both concentrations of H2O2 elevated Hsp-70 protein levels, but more so in 1000 than 100 µM H2O2 (aii). RS (10 and 20 µM) pre-conditioning attenuated the effects of 1000 µM H2O2 on Hsp-70 (aii), but not on p66Shc-Ser36 phosphorylation (ai). DM vs. CT: in none of the cases was significant. P-values calculated using a two-tailed Student's t-test.*: P<0.01 vs CT; ¤: P<0.001 vs H2O2 1000 µM; ‡: P<0.01 vs H2O2 100 µM. Data are mean ± s.e.m.

Mentions: The effects of H2O2 on mitochondrial membrane potential (ΔΨm) and its implication in the activation of the mitochondrial apoptosis cascade are well recognized37. Resveratrol has been suggested to counteract apoptosis induced by H2O2. Therefore, we analysed whether RS was able to counteract H2O2-induced mitochondria depolarisation. RS pre-conditioning did not prevent, but even enhanced the depolarization induced by 24 h exposure to 1000 µM H2O2 (Fig. 7). Notably, this was associated with increased phosphorylation of p66Shc(A)-Ser36 (Fig. 8ai), and elevated Hsp-70 protein levels (Fig. 8aii), two key players in the mitochondrial and cellular stress response3738. In particular, 1000 µM, but not 100 µM-H2O2 induced p66Shc(A)-Ser36 phosphorylation (Fig. 8ai) and both elevated Hsp-70 protein levels (Fig. 8aii). Although pre-incubation with RS reduced the 1000 µM H2O2-induced elevation in Hsp70, it did not alleviate the increased p66Shc(A) phosphorylation.


The impact of resveratrol and hydrogen peroxide on muscle cell plasticity shows a dose-dependent interaction.

Bosutti A, Degens H - Sci Rep (2015)

Resveratrol induced p66Shc-Ser36 phosphorylation and up-regulation of Heat Shock Protein (Hsp)-70.(a) Western blot analysis showing the effect of resveratrol (RS; 10, 20, 40 and 60 µM), H2O2 (100 and 1000 µM) and RS pre-conditioning (10 and 20 µM) on p66Shc-Ser36 phosphorylation and Hsp-70 protein levels. Dose dependent increases in Ser36-p66 phosphorylation and in Hsp-70 protein contents were observed in resveratrol-treated cells. 1000 µM, but not 100 µM H2O2 induced p66Shc(A)-Ser36 phosphorylation (ai); both concentrations of H2O2 elevated Hsp-70 protein levels, but more so in 1000 than 100 µM H2O2 (aii). RS (10 and 20 µM) pre-conditioning attenuated the effects of 1000 µM H2O2 on Hsp-70 (aii), but not on p66Shc-Ser36 phosphorylation (ai). DM vs. CT: in none of the cases was significant. P-values calculated using a two-tailed Student's t-test.*: P<0.01 vs CT; ¤: P<0.001 vs H2O2 1000 µM; ‡: P<0.01 vs H2O2 100 µM. Data are mean ± s.e.m.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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f8: Resveratrol induced p66Shc-Ser36 phosphorylation and up-regulation of Heat Shock Protein (Hsp)-70.(a) Western blot analysis showing the effect of resveratrol (RS; 10, 20, 40 and 60 µM), H2O2 (100 and 1000 µM) and RS pre-conditioning (10 and 20 µM) on p66Shc-Ser36 phosphorylation and Hsp-70 protein levels. Dose dependent increases in Ser36-p66 phosphorylation and in Hsp-70 protein contents were observed in resveratrol-treated cells. 1000 µM, but not 100 µM H2O2 induced p66Shc(A)-Ser36 phosphorylation (ai); both concentrations of H2O2 elevated Hsp-70 protein levels, but more so in 1000 than 100 µM H2O2 (aii). RS (10 and 20 µM) pre-conditioning attenuated the effects of 1000 µM H2O2 on Hsp-70 (aii), but not on p66Shc-Ser36 phosphorylation (ai). DM vs. CT: in none of the cases was significant. P-values calculated using a two-tailed Student's t-test.*: P<0.01 vs CT; ¤: P<0.001 vs H2O2 1000 µM; ‡: P<0.01 vs H2O2 100 µM. Data are mean ± s.e.m.
Mentions: The effects of H2O2 on mitochondrial membrane potential (ΔΨm) and its implication in the activation of the mitochondrial apoptosis cascade are well recognized37. Resveratrol has been suggested to counteract apoptosis induced by H2O2. Therefore, we analysed whether RS was able to counteract H2O2-induced mitochondria depolarisation. RS pre-conditioning did not prevent, but even enhanced the depolarization induced by 24 h exposure to 1000 µM H2O2 (Fig. 7). Notably, this was associated with increased phosphorylation of p66Shc(A)-Ser36 (Fig. 8ai), and elevated Hsp-70 protein levels (Fig. 8aii), two key players in the mitochondrial and cellular stress response3738. In particular, 1000 µM, but not 100 µM-H2O2 induced p66Shc(A)-Ser36 phosphorylation (Fig. 8ai) and both elevated Hsp-70 protein levels (Fig. 8aii). Although pre-incubation with RS reduced the 1000 µM H2O2-induced elevation in Hsp70, it did not alleviate the increased p66Shc(A) phosphorylation.

Bottom Line: RS did not increase oxidative capacity.In conclusion, low resveratrol doses promoted in vitro muscle regeneration and attenuated the impact of ROS, while high doses augmented the reduced plasticity and metabolism induced by oxidative stress.Thus, the effects of resveratrol depend on its dose and degree of oxidative stress.

View Article: PubMed Central - PubMed

Affiliation: School of Healthcare Science, Manchester Metropolitan University, Manchester, United Kingdom.

ABSTRACT
While reactive oxygen species (ROS) play a role in muscle repair, excessive amounts of ROS for extended periods may lead to oxidative stress. Antioxidants, as resveratrol (RS), may reduce oxidative stress, restore mitochondrial function and promote myogenesis and hypertrophy. However, RS dose-effectiveness for muscle plasticity is unclear. Therefore, we investigated RS dose-response on C2C12 myoblast and myotube plasticity 1. in the presence and 2. absence of different degrees of oxidative stress. Low RS concentration (10 μM) stimulated myoblast cell cycle arrest, migration and sprouting, which were inhibited by higher doses (40-60 μM). RS did not increase oxidative capacity. In contrast, RS induced mitochondria loss, reduced cell viability and ROS production, and activated stress response pathways [Hsp70 and pSer36-p66(ShcA) proteins]. However, the deleterious effects of H2O2 (1000 µM) on cell migration were alleviated after preconditioning with 10 µM-RS. This dose also enhanced cell motility mediated by 100 µM-H2O2, while higher RS-doses augmented the H2O2-induced impaired myoblast regeneration and mitochondrial dehydrogenase activity. In conclusion, low resveratrol doses promoted in vitro muscle regeneration and attenuated the impact of ROS, while high doses augmented the reduced plasticity and metabolism induced by oxidative stress. Thus, the effects of resveratrol depend on its dose and degree of oxidative stress.

Show MeSH
Related in: MedlinePlus