The present study aimed to investigate the effects of Laspartic acid potassium salt (potassium aspartate, Kasp) on SHSY5Y cells treated with ouabain and H2O2. An 3(4,5dimethylthiazol2yl)2,5diphenyltetrazolium bromide (MTT) assay was performed to investigate the effects of Kasp on SHSY5Y
cell death induced by ouabain. Nissl staining was used to demonstrate the morphological changes of the SHSY5Y cells. Light microscopy and 4',6diamidino2phenylindole (DAPI) staining were performed to visualize apoptosis in SHSY5Y cells incubated with ouabain for 6, 24 and 48 h. Transmission
electron microscopy was used to observe the effect of Kasp on ultrastructural changes of the SHSY5Y cells following incubation with ouabain for 24 and 48 h. An annexin Vfluorescein isothiocyanate/propidium binding assay and flow cytometry were performed successively to investigate
how Kasp affected the H2O2induced cell apoptosis. The MTT assay demonstrated that Kasp attenuated the cytotoxicity of the SHSY5Y cells following treatment with ouabain, in a dosedependent manner. The cell survival rates following 48 h incubation in the Kasp (15 mM) and Kasp (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 Kasp (25 mM) groups were alleviated. In the DAPI staining and transmission electron microscopy analyses, KCl and Kasp (25 mM) reduced the rate of
ouabaininduced apoptosis. Flow cytometry revealed that Kasp (25 mM) reduced H2O2induced apoptosis. These results demonstrated that Kasp (25 mM) inhibited the ouabain and H2O2induced SHSY5Y cell damage and apoptosis, possibly by supplementing levels of intracellular K+.
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Document Type: Research Article
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
Publication date: August 1, 2015
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Molecular Medicine Reports is a monthly, peer-reviewed journal available in print and online, that includes studies devoted to molecular medicine, underscoring aspects including pharmacology, pathology, genetics, neurosciences, infectious diseases, molecular cardiology and molecular surgery. In vitro and in vivo studies of experimental model systems pertaining to the mechanisms of a variety of diseases offer researchers the necessary tools and knowledge with which to aid the diagnosis and treatment of human diseases.
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