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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 (10 µM) prevented the deleterious effect of H2O2 on cell migration but not cell fusion.(a) Phase contrast images showing the effect of RS alone (10 and 20 µM) on cell migration (24 h). (b, c) Phase contrast images showing the effects of different pre-conditioning resveratrol (RS) concentrations and H2O2 on cell migration (b) and fusion (c). The number of migrated cells and myotubes in the different conditions are summarized in (d) and (e), respectively. (b and graph in d) show that the effects of 24 h H2O2 on cell migration were dose-dependent. High doses of H2O2 (500–1000 µM) blocked cell motility, while it was increased with 100 µM H2O2. (b and graph in d) shows that RS pre-conditioning (10 µM) abolished the deleterious effects of 500 µM, attenuated that of 1000 µM H2O2 and further enhanced cell migration induced by H2O2 100 µM. (c,e) 500 and 1000 µM H2O2 did block cell fusion, which was not rescued with RS preconditioning. Data are expressed as mean ± s.e.m of biological quadruplicate. *: P<0.01 vs. CT; ‡: P<0.01 vs. RS 10 µM; ¤: P<0.01 vs. RS 20 µM. P-value calculated using a two-tailed Student's t-test. Bars 20 µm. Original magnification, x 50.
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f5: Resveratrol (10 µM) prevented the deleterious effect of H2O2 on cell migration but not cell fusion.(a) Phase contrast images showing the effect of RS alone (10 and 20 µM) on cell migration (24 h). (b, c) Phase contrast images showing the effects of different pre-conditioning resveratrol (RS) concentrations and H2O2 on cell migration (b) and fusion (c). The number of migrated cells and myotubes in the different conditions are summarized in (d) and (e), respectively. (b and graph in d) show that the effects of 24 h H2O2 on cell migration were dose-dependent. High doses of H2O2 (500–1000 µM) blocked cell motility, while it was increased with 100 µM H2O2. (b and graph in d) shows that RS pre-conditioning (10 µM) abolished the deleterious effects of 500 µM, attenuated that of 1000 µM H2O2 and further enhanced cell migration induced by H2O2 100 µM. (c,e) 500 and 1000 µM H2O2 did block cell fusion, which was not rescued with RS preconditioning. Data are expressed as mean ± s.e.m of biological quadruplicate. *: P<0.01 vs. CT; ‡: P<0.01 vs. RS 10 µM; ¤: P<0.01 vs. RS 20 µM. P-value calculated using a two-tailed Student's t-test. Bars 20 µm. Original magnification, x 50.

Mentions: Our second objective was to establish the optimal RS concentration to counteract or synergise the effects of H2O2 on cell migration and cell fusion (Fig. 5). We found that the effects of 24 h H2O2 on cell migration were dose-dependent (Fig. 5b,d). High doses of H2O2 (500 and 1000 µM) blocked cell motility almost completely, while it was increased by 100 µM H2O2, (Fig. 5b,d). High doses of H2O2 (500 and 1000 µM) also blocked cell fusion, as indicated by a reduced number of myotubes (Fig. 5c,e). Notably, 24 h of 10 µM RS pre-conditioning abolished the deleterious effects of 500 µM on cell migration and attenuated that of 1000 µM H2O2. In addition, 10 µM RS further enhanced cell migration induced by 100 µM H2O2. Twenty µM RS did not enhance the stimulating effect of 100 µM H2O2 nor have additional protective effects (Fig. 5b,d). Preconditioning with 10 µM RS for 24 h did, however, not rescue the inhibition on cell fusion induced by 1000 µM H2O2 (Fig. 5c,e).


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

Bosutti A, Degens H - Sci Rep (2015)

Resveratrol (10 µM) prevented the deleterious effect of H2O2 on cell migration but not cell fusion.(a) Phase contrast images showing the effect of RS alone (10 and 20 µM) on cell migration (24 h). (b, c) Phase contrast images showing the effects of different pre-conditioning resveratrol (RS) concentrations and H2O2 on cell migration (b) and fusion (c). The number of migrated cells and myotubes in the different conditions are summarized in (d) and (e), respectively. (b and graph in d) show that the effects of 24 h H2O2 on cell migration were dose-dependent. High doses of H2O2 (500–1000 µM) blocked cell motility, while it was increased with 100 µM H2O2. (b and graph in d) shows that RS pre-conditioning (10 µM) abolished the deleterious effects of 500 µM, attenuated that of 1000 µM H2O2 and further enhanced cell migration induced by H2O2 100 µM. (c,e) 500 and 1000 µM H2O2 did block cell fusion, which was not rescued with RS preconditioning. Data are expressed as mean ± s.e.m of biological quadruplicate. *: P<0.01 vs. CT; ‡: P<0.01 vs. RS 10 µM; ¤: P<0.01 vs. RS 20 µM. P-value calculated using a two-tailed Student's t-test. Bars 20 µm. Original magnification, x 50.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4308712&req=5

f5: Resveratrol (10 µM) prevented the deleterious effect of H2O2 on cell migration but not cell fusion.(a) Phase contrast images showing the effect of RS alone (10 and 20 µM) on cell migration (24 h). (b, c) Phase contrast images showing the effects of different pre-conditioning resveratrol (RS) concentrations and H2O2 on cell migration (b) and fusion (c). The number of migrated cells and myotubes in the different conditions are summarized in (d) and (e), respectively. (b and graph in d) show that the effects of 24 h H2O2 on cell migration were dose-dependent. High doses of H2O2 (500–1000 µM) blocked cell motility, while it was increased with 100 µM H2O2. (b and graph in d) shows that RS pre-conditioning (10 µM) abolished the deleterious effects of 500 µM, attenuated that of 1000 µM H2O2 and further enhanced cell migration induced by H2O2 100 µM. (c,e) 500 and 1000 µM H2O2 did block cell fusion, which was not rescued with RS preconditioning. Data are expressed as mean ± s.e.m of biological quadruplicate. *: P<0.01 vs. CT; ‡: P<0.01 vs. RS 10 µM; ¤: P<0.01 vs. RS 20 µM. P-value calculated using a two-tailed Student's t-test. Bars 20 µm. Original magnification, x 50.
Mentions: Our second objective was to establish the optimal RS concentration to counteract or synergise the effects of H2O2 on cell migration and cell fusion (Fig. 5). We found that the effects of 24 h H2O2 on cell migration were dose-dependent (Fig. 5b,d). High doses of H2O2 (500 and 1000 µM) blocked cell motility almost completely, while it was increased by 100 µM H2O2, (Fig. 5b,d). High doses of H2O2 (500 and 1000 µM) also blocked cell fusion, as indicated by a reduced number of myotubes (Fig. 5c,e). Notably, 24 h of 10 µM RS pre-conditioning abolished the deleterious effects of 500 µM on cell migration and attenuated that of 1000 µM H2O2. In addition, 10 µM RS further enhanced cell migration induced by 100 µM H2O2. Twenty µM RS did not enhance the stimulating effect of 100 µM H2O2 nor have additional protective effects (Fig. 5b,d). Preconditioning with 10 µM RS for 24 h did, however, not rescue the inhibition on cell fusion induced by 1000 µM H2O2 (Fig. 5c,e).

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