Effect of Creatine on the Survival of RGC-5 Cells under Serum Deprivation

Jae Woo Kim; Jung Heum Hong; Sun Hee Kang; Yun Young Kim

doi:10.3341/jkos.2011.52.5.618

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Kim, Hong, Kang, and Kim: Effect of Creatine on the Survival of RGC-5 Cells under Serum Deprivation

Original Article

J Korean Ophthalmol Soc 2011;52(5):618-623.

Published online: 3 August 2011

DOI: https://doi.org/10.3341/jkos.2011.52.5.618

Effect of Creatine on the Survival of RGC-5 Cells under Serum Deprivation

Jae Woo Kim, MD, PhD, Jung Heum Hong, MD, Sun Hee Kang, MD, Yun Young Kim, MD, PhD

Department of Ophthalmology, Catholic University of Daegu School of Medicine, Daegu, Korea

Address reprint requests to Jae Woo Kim, MD, PhD Department of Ophthalmology, Daegu Catholic University Medical Center #3056-6 Daemyeong 4-dong, Nam-gu, Daegu 705-718, Korea Tel: 82-53-650-4728, Fax: 82-53-627-0133, E-mail: jwkim@cu.ac.kr

Received 13 August 20109 November 2010 Accepted 8 March 2011

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Purpose

To evaluate the protective effect of creatine on the survival of retinal ganglion cells after serum deprivation.

Methods

RGC-5 cells were exposed to 5 mM creatine with serum-free media for 4 days. Cellular survival and mitochondrial respiratory activity were measured with MTT assay and resazurin assay, respectively. Degree of apoptosis was evaluated with vital staining using acridine orange/Hoechest 33342 and flow cytometric analysis using annexin/PI, respectively.

Results

Creatine increased cellular survival of RGC-5 cells significantly after serum deprivation. Additionally, creatine increased mitochondrial respiratory activity and inhibited apoptosis of RGC-5 cells.

Conclusions

The energy precursor creatine increased survival of retinal ganglion cells after serum deprivation. Creatine could be relevant for the cytoprotection of retinal ganglion cells.

Keywords: Creatine, Cytoprotection, Retinal ganglion cells

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Figure 1.

Effect of creatine on the survival of RGC-5 cells after serum deprivation for 4 days. Addition of 5 mM creatine significantly increased cellular survival. ^* p < 0.05.

Figure 2.

AO/HO double staining of RGC-5 cells with or without treatment of 5 mM creatine for 4 days. Normal control cell stains blue nuclei and brilliant red cytoplasm. In contrast, the apoptotic cell (arrows) shows yellow to orange-red nuclei, becoming indis-tinguishable from the cytoplasm. Addition of 5 mM creatine (A) revealed less apoptotic cells compared non-treated control (B) (×200).

Figure 3.

Flow cytometric analysis of the effect of creatine on the survival of RGC-5 cells after serum deprivation for 4 days. Addition of 5 mM creatine significantly decreased cellular apoptosis. ^* p < 0.05.

Figure 4.

Effect of creatine on the mitochondrial respiratory activity in the RGC-5 cells after serum deprivation for 4 days. Addition of 5 mM creatine significantly increased mitochondrial respiratory activity compared to non-treated controls.

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