Acute Electrical Pulse Stimulation and Hyperglycemia Regulates RCAN1-4 in C2C12 myotubes through Oxidative Stress

Ramin Emrani Bidi; Amelie Rebillard; Dany Saligaut; Arlette Delamarche; Kelvin J A Davies; Josiane Cillard

doi:10.1016/j.freeradbiomed.2014.10.757

Acute Electrical Pulse Stimulation and Hyperglycemia Regulates RCAN1-4 in C2C12 myotubes through Oxidative Stress

Free Radic Biol Med. 2014 Oct:75 Suppl 1:S29. doi: 10.1016/j.freeradbiomed.2014.10.757. Epub 2014 Dec 10.

Authors

Ramin Emrani Bidi¹, Amelie Rebillard², Dany Saligaut³, Arlette Delamarche², Kelvin J A Davies⁴, Josiane Cillard³

Affiliations

¹ EA1274 (Faculté de pharmacie), Université de Rennes 1, France. Electronic address: ramin.emrani@gmail.com.
² EA1274 (M2S), Université de Rennes 2, France.
³ EA1274 (Faculté de pharmacie), Université de Rennes 1, France.
⁴ Ethel Percy Andrus Gerontology Center of the Davis School of Gerontology; and Division of Molecular and Computational Biology (Department of Biological Sciences, of the Dornsife College of Letters, Arts & Sciences), the University of Southern California,Los Angeles, California 90089-0191, USA.

PMID: 26461328
DOI: 10.1016/j.freeradbiomed.2014.10.757

Abstract

Regulator of Calcineurin1 (RCAN1) controls the Sr/Thr phosphatase, Calcineurin. RCAN1 has never been studied in-vitro in myotubes following acute Electrical Pulse Stimulation (EPS) which recapitulates the physiological effect of exhaustive exercise and hyperglycemia (HG). 144 h differentiated C2C12 myotubes were treated either with high glucose (HG) at 15mM or kept as control for 48 h. The myotubes were subjected to EPS for 1second followed by 1second of pause for 90min. Two separate Non-EPS controls with and without HG were performed simultaneously. The protein level of RCAN1-4 increased immediately after EPS up to 6hours in HG myotubes and in control. Non-EPS myotubes treated with HG exhibited a higher level of RCAN1-4 compare to Non-EPS control. RCAN1-1L and 1-S did not show any significant change after EPS in both groups. Calcineurin level decreased immediately after EPS. The levels of CuZn SOD and MnSOD were increased immediately after EPS whereas Catalase level increased significantly only 3hours after EPS in both groups. CuZn SOD and MnSOD showed higher level in HG control compare to control while the level of Catalase was lesser in HG control. NFATc4 level decreased at 3Hr post EPS in both groups. PGC1-? remained unchanged in all the samples. 4HNE increased significantly in both groups after EPS and was higher in HG control than in control. Protein Carbonyl (PC) increased significantly 3hours after EPS in normal cells and returned to basal level at 6 h. However in HG myotubes, PC level increased immediately after EPS and no further modification until 6Hr was observed. HG samples exhibited much higher PC than normal samples. We have already shown than exhaustive exercise regulates RCAN1-4 in rat skeletal muscle through involvement of oxidative stress. The acute EPS of C2C12 myotubes in normal condition or under hyperglycemia, served as an in-vitro model of exercise and diabetic regulation of RCAN1 through oxidative stress in-vitro.