Use of Creatine and Creatinine to Minimize Doxorubicin-Induced Cytotoxicity in Cardiac and Skeletal Muscle Myoblasts

Abstract

Doxorubicin (DOX), an effective anticancer agent, can damage cardiac and skeletal muscle tissue via excessive generation of reactive oxygen species (ROS). Supplemental creatine (Cr) has been shown to have a therapeutic role in disease states characterized by increased oxidative stress. To investigate the effects of Cr and creatinine (CrN) on DOX-induced cytotoxicity. Cultured L6 and H9C2 myoblasts were exposed to 25 μM DOX, 10 mM Cr, 10 mM CrN, 25 μM DOX + 10 mM Cr, 25 μM DOX + 10 mM CrN, or control media for 12 h. Viability was assessed using Confocal and Widefield imaging. Immunoblotting was used to determine protein expression. Viability was lowest in the DOX-treated group regardless of cell type; however, when DOX was combined with Cr or CrN, viability was improved. Levels of oxidative stress, as measured by 4-hydroxynonenal (4HNE), were significantly (p < 0.05) higher in the DOX treated cells vs. controls; however, Cr + DOX and CrN + DOX significantly lowered 4HNE levels compared to DOX-treated cells. Creatine kinase (CK), a key marker of cellular damage, was significantly higher in DOX-treated H9c2 cells vs. controls. However, Cr or CrN in combination with DOX, resulted in no significant differences in CK vs. controls. Supplementation with Cr or CrN may preserve cell viability during DOX treatment.

Disclaimer

The views expressed in the submitted article are his own and not the official position of the institution or funder.

Conflict of Interest

In accordance with Taylor & Francis policy, no potential conflict of interests are reported by the authors.

Author Contributions

The experiment was designed by Eric Bredahl. Eric Bredahl, Wisam Najadawi, and Caroline Pass performed the research. Eric Bredahl performed the statistics. Eric Bredahl, Jake Siedlik, Joan Eckerson, and Kristen Drescher wrote and reviewed the manuscript.

Additional information

Funding

This study was supported by a George F. Haddix research grant awarded by Creighton University. The data were collected at the Integrated Biological Imaging Facility at Creighton University, Omaha, NE. This facility is supported by the Creighton University School of Medicine and grants GM103427 and GM110768 from the National Institute of General Medical Science (NIGMS), a component of the National Institutes of Health (NIH). The facility was constructed with support from grants from the National Center for Research Resources (RR016469) and the NIGMS (GM103427). This investigation is solely the responsibility of the authors and does not necessarily represent the official views of NIGMS or NIH.