Investigation of the Effect of Melatonin Administration on Calpain-1 Expression and Atrophic Morphology in Atrophy of Mouse Myoblast Cell Line Exposed to Hydrogen Peroxide

Karimi N., Kartal Y., Budak M. T.

"Türk Fizyolojik Bilimler Derneği 45. Ulusal Fizyoloji Kongresi", Aydın, Turkey, 31 October - 03 November 2019, pp.34

  • Publication Type: Conference Paper / Summary Text
  • Doi Number: 10.1111/apha.13423
  • City: Aydın
  • Country: Turkey
  • Page Numbers: pp.34
  • Hacettepe University Affiliated: Yes

Abstract

It is known that oxidative stress is the main factor in the formation of disuse muscle atrophy, the most common type of muscle atrophy. The first important damage which is exerted by oxidative stress on proteases such as calpain-1 has been demonstrated in several studies. Treatment options include antioxidants, and in these groups mitochondrial targeting antioxidants such as melatonin have been shown to be more effective.

AIM: In this study, we aimed to determine the effect of melatonin treatment on oxidative stress-induced atrophy morphological change, calpain-1 expression and redox balance in C2C12 muscle cells.

METHOD: We have used H2O2 to generate oxidative stress. Four groups of C2C12 cells were created as K(Control), M(Melatonin), H(H2O2 ), M+H (Melatonin+ H2O2 ) groups. For morphological evaluation, measurement of myotube diameter with Image J Program was performed. In the second step, the amount of calpain-1 protein from total protein lysates was measured by western blot method. TAS and TOS kits were used to find out the antioxidant/oxidant statuses of the cells. Statistical analyzes were performed in SPSS program. The level of significance was set at p <0.05.

RESULTS: There was a statistically significant decrease in the mean myotube diameters of the H group compared to the other groups (K, M, M+H) and statistically significant increase in mean myotube diameter of M group compared to K and H groups. The expression level of calpain-1 protein was increased in H, M and M+H groups compared with K group. TOS was higher in the H group than other groups. The highest mean TAS value was seen in the H group (rebound effect).

CONCLUSION: These results suggest that H2O2 produces muscle atrophy in C2C12 cells and that melatonin prevents morphological change and in addition to calpain-1 protein, other mechanisms can also take part in the prevention of disuse muscle atrophy. Functional studies need to be carried out to better explain this effect.