Potassium aspartate inhibits SH‑SY5Y cell damage and apoptosis induced by ouabain and H2O2

Sun,Xuefei; Min,Dongyu; Wang,Yan; Hao,Liying

doi:10.3892/mmr.2015.3741

Potassium aspartate inhibits SH‑SY5Y cell damage and apoptosis induced by ouabain and H2O2

Authors:
- Xuefei Sun
- Dongyu Min
- Yan Wang
- Liying Hao
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Affiliations: Department of Pharmaceutical Toxicology, School of Pharmaceutical Science, China Medical University, Shenyang, Liaoning 110001, P.R. China, The Experimental Center of Traditional Chinese Medicine, The Affiliated Hospital of Liaoning University of Traditional Chinese Medicine, Shenyang, Liaoning 110032, P.R. China
Published online on: May 7, 2015 https://doi.org/10.3892/mmr.2015.3741
Pages: 2842-2848

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Abstract

The present study aimed to investigate the effects of L‑aspartic acid potassium salt (potassium aspartate, K‑asp) on SH‑SY5Y cells treated with ouabain and H2O2. An 3‑(4,5‑dimethylthiazol‑2‑yl)‑2,5‑diphenyltetrazolium bromide (MTT) assay was performed to investigate the effects of K‑asp on SH‑SY5Y cell death induced by ouabain. Nissl staining was used to demonstrate the morphological changes of the SH‑SY5Y cells. Light microscopy and 4',6‑diamidino‑2‑phenylindole (DAPI) staining were performed to visualize apoptosis in SH‑SY5Y cells incubated with ouabain for 6, 24 and 48 h. Transmission electron microscopy was used to observe the effect of K‑asp on ultrastructural changes of the SH‑SY5Y cells following incubation with ouabain for 24 and 48 h. An annexin V‑fluorescein isothiocyanate/propidium binding assay and flow cytometry were performed successively to investigate how K‑asp affected the H2O2‑induced cell apoptosis. The MTT assay demonstrated that K‑asp attenuated the cytotoxicity of the SH‑SY5Y cells following treatment with ouabain, in a dose‑dependent manner. The cell survival rates following 48 h incubation in the K‑asp (15 mM) and K‑asp (25 mM) groups were higher compared with the KCl and MK801 groups. Nissl staining demonstrated that the severity of cell injury in the KCl and K‑asp (25 mM) groups were alleviated. In the DAPI staining and transmission electron microscopy analyses, KCl and K‑asp (25 mM) reduced the rate of ouabain‑induced apoptosis. Flow cytometry revealed that K‑asp (25 mM) reduced H2O2‑induced apoptosis. These results demonstrated that K‑asp (25 mM) inhibited the ouabain and H2O2‑induced SH‑SY5Y cell damage and apoptosis, possibly by supplementing levels of intracellular K+.

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