Limits...
Low-Level Laser Therapy (LLLT) in Dystrophin-Deficient Muscle Cells: Effects on Regeneration Capacity, Inflammation Response and Oxidative Stress.

Macedo AB, Moraes LH, Mizobuti DS, Fogaça AR, Moraes Fdos S, Hermes Tde A, Pertille A, Minatel E - PLoS ONE (2015)

Bottom Line: The mdx LA 48 group showed significant increase in Myosin Heavy Chain levels compared to the untreated mdx and mdx LA 24 groups.The mdx LA 24 and mdx LA 48 groups showed significant increase in [Ca2+]i.Together, these results suggest that the laser treatment improved regenerative capacity and decreased inflammatory response and oxidative stress in dystrophic muscle cells, indicating that LLLT could be a helpful alternative therapy to be associated with other treatment for dystrophinopathies.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Biologia Estrutural e Funcional, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), Campinas, SP, Brazil.

ABSTRACT
The present study evaluated low-level laser therapy (LLLT) effects on some physiological pathways that may lead to muscle damage or regeneration capacity in dystrophin-deficient muscle cells of mdx mice, the experimental model of Duchenne muscular dystrophy (DMD). Primary cultures of mdx skeletal muscle cells were irradiated only one time with laser and analyzed after 24 and 48 hours. The LLLT parameter used was 830 nm wavelengths at 5 J/cm² fluence. The following groups were set up: Ctrl (untreated C57BL/10 primary muscle cells), mdx (untreated mdx primary muscle cells), mdx LA 24 (mdx primary muscle cells - LLLT irradiated and analyzed after 24 h), and mdx LA 48 (mdx primary muscle cells - LLLT irradiated and analyzed after 48 h). The mdx LA 24 and mdx LA 48 groups showed significant increase in cell proliferation, higher diameter in muscle cells and decreased MyoD levels compared to the mdx group. The mdx LA 48 group showed significant increase in Myosin Heavy Chain levels compared to the untreated mdx and mdx LA 24 groups. The mdx LA 24 and mdx LA 48 groups showed significant increase in [Ca2+]i. The mdx group showed significant increase in H2O2 production and 4-HNE levels compared to the Ctrl group and LLLT treatment reduced this increase. GSH levels and GPx, GR and SOD activities increased in the mdx group. Laser treatment reduced the GSH levels and GR and SOD activities in dystrophic muscle cells. The mdx group showed significant increase in the TNF-α and NF-κB levels, which in turn was reduced by the LLLT treatment. Together, these results suggest that the laser treatment improved regenerative capacity and decreased inflammatory response and oxidative stress in dystrophic muscle cells, indicating that LLLT could be a helpful alternative therapy to be associated with other treatment for dystrophinopathies.

No MeSH data available.


Related in: MedlinePlus

Analysis of inflammation in control and dystrophic muscle cells.Immunoblot analysis of TNF-α (A) and NF-κB (B). Graphs show protein level in the muscle cells from normal (Ctrl) and dystrophic primary muscle cells untreated (mdx untreated) and LLLT treatment analyzed after 24 hours (mdx LA 24) and 48 hours (mdx LA 48). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a loading control. All the experiments were performed in triplicate, and the relative value of the band intensity was quantified and normalized by the corresponding Ctrl. a P< 0.05 versus Ctrl; b P< 0.05 versus mdx untreated; c P< 0.05 versus mdx LA 24. Error bars, SD.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4470633&req=5

pone.0128567.g005: Analysis of inflammation in control and dystrophic muscle cells.Immunoblot analysis of TNF-α (A) and NF-κB (B). Graphs show protein level in the muscle cells from normal (Ctrl) and dystrophic primary muscle cells untreated (mdx untreated) and LLLT treatment analyzed after 24 hours (mdx LA 24) and 48 hours (mdx LA 48). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a loading control. All the experiments were performed in triplicate, and the relative value of the band intensity was quantified and normalized by the corresponding Ctrl. a P< 0.05 versus Ctrl; b P< 0.05 versus mdx untreated; c P< 0.05 versus mdx LA 24. Error bars, SD.

Mentions: Inflammatory response of laser therapy was analyzed by the TNF-α and NF-κB levels (Fig 5A and 5B). Dystrophic primary muscle cells presented high levels of TNF-α and NF-κB (by 15% and 22%, respectively) compared to the control. The mdx LA 24 and mdx LA 48 primary muscle cells showed a significant reduction (by 8% and 9%, respectively) on TNF-α levels compared to control. In addition, the mdx LA 48 group showed a significant decrease on NF-κB levels (by 25% and 14%, respectively) compared to Ctrl and mdx LA 24 groups.


Low-Level Laser Therapy (LLLT) in Dystrophin-Deficient Muscle Cells: Effects on Regeneration Capacity, Inflammation Response and Oxidative Stress.

Macedo AB, Moraes LH, Mizobuti DS, Fogaça AR, Moraes Fdos S, Hermes Tde A, Pertille A, Minatel E - PLoS ONE (2015)

Analysis of inflammation in control and dystrophic muscle cells.Immunoblot analysis of TNF-α (A) and NF-κB (B). Graphs show protein level in the muscle cells from normal (Ctrl) and dystrophic primary muscle cells untreated (mdx untreated) and LLLT treatment analyzed after 24 hours (mdx LA 24) and 48 hours (mdx LA 48). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a loading control. All the experiments were performed in triplicate, and the relative value of the band intensity was quantified and normalized by the corresponding Ctrl. a P< 0.05 versus Ctrl; b P< 0.05 versus mdx untreated; c P< 0.05 versus mdx LA 24. Error bars, SD.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4470633&req=5

pone.0128567.g005: Analysis of inflammation in control and dystrophic muscle cells.Immunoblot analysis of TNF-α (A) and NF-κB (B). Graphs show protein level in the muscle cells from normal (Ctrl) and dystrophic primary muscle cells untreated (mdx untreated) and LLLT treatment analyzed after 24 hours (mdx LA 24) and 48 hours (mdx LA 48). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as a loading control. All the experiments were performed in triplicate, and the relative value of the band intensity was quantified and normalized by the corresponding Ctrl. a P< 0.05 versus Ctrl; b P< 0.05 versus mdx untreated; c P< 0.05 versus mdx LA 24. Error bars, SD.
Mentions: Inflammatory response of laser therapy was analyzed by the TNF-α and NF-κB levels (Fig 5A and 5B). Dystrophic primary muscle cells presented high levels of TNF-α and NF-κB (by 15% and 22%, respectively) compared to the control. The mdx LA 24 and mdx LA 48 primary muscle cells showed a significant reduction (by 8% and 9%, respectively) on TNF-α levels compared to control. In addition, the mdx LA 48 group showed a significant decrease on NF-κB levels (by 25% and 14%, respectively) compared to Ctrl and mdx LA 24 groups.

Bottom Line: The mdx LA 48 group showed significant increase in Myosin Heavy Chain levels compared to the untreated mdx and mdx LA 24 groups.The mdx LA 24 and mdx LA 48 groups showed significant increase in [Ca2+]i.Together, these results suggest that the laser treatment improved regenerative capacity and decreased inflammatory response and oxidative stress in dystrophic muscle cells, indicating that LLLT could be a helpful alternative therapy to be associated with other treatment for dystrophinopathies.

View Article: PubMed Central - PubMed

Affiliation: Departamento de Biologia Estrutural e Funcional, Instituto de Biologia, Universidade Estadual de Campinas (UNICAMP), Campinas, SP, Brazil.

ABSTRACT
The present study evaluated low-level laser therapy (LLLT) effects on some physiological pathways that may lead to muscle damage or regeneration capacity in dystrophin-deficient muscle cells of mdx mice, the experimental model of Duchenne muscular dystrophy (DMD). Primary cultures of mdx skeletal muscle cells were irradiated only one time with laser and analyzed after 24 and 48 hours. The LLLT parameter used was 830 nm wavelengths at 5 J/cm² fluence. The following groups were set up: Ctrl (untreated C57BL/10 primary muscle cells), mdx (untreated mdx primary muscle cells), mdx LA 24 (mdx primary muscle cells - LLLT irradiated and analyzed after 24 h), and mdx LA 48 (mdx primary muscle cells - LLLT irradiated and analyzed after 48 h). The mdx LA 24 and mdx LA 48 groups showed significant increase in cell proliferation, higher diameter in muscle cells and decreased MyoD levels compared to the mdx group. The mdx LA 48 group showed significant increase in Myosin Heavy Chain levels compared to the untreated mdx and mdx LA 24 groups. The mdx LA 24 and mdx LA 48 groups showed significant increase in [Ca2+]i. The mdx group showed significant increase in H2O2 production and 4-HNE levels compared to the Ctrl group and LLLT treatment reduced this increase. GSH levels and GPx, GR and SOD activities increased in the mdx group. Laser treatment reduced the GSH levels and GR and SOD activities in dystrophic muscle cells. The mdx group showed significant increase in the TNF-α and NF-κB levels, which in turn was reduced by the LLLT treatment. Together, these results suggest that the laser treatment improved regenerative capacity and decreased inflammatory response and oxidative stress in dystrophic muscle cells, indicating that LLLT could be a helpful alternative therapy to be associated with other treatment for dystrophinopathies.

No MeSH data available.


Related in: MedlinePlus