Study of the protective effect of calcium channel blockers against neuronal damage induced by glutamate in cultured hippocampal neurons

• Published in: Pharmacological reports Vol. 65; no. 3; pp. 730 - 736
• Main Authors: Sendrowski, Krzysztof, Rusak, Małgorzata, Sobaniec, Piotr, Iłendo, Elżbieta, Dąbrowska, Milena, Boćkowski, Leszek, Koput, Alicja, Sobaniec, Wojciech
• Format: Journal Article
• Language: English
• Published: Cham Springer International Publishing 2013
• Subjects: Animals; Apoptosis - drug effects; Calcium Channel Blockers - pharmacology; Cell Death - drug effects; Cells, Cultured; Drug Safety and Pharmacovigilance; Glutamic Acid - adverse effects; Glutamic Acid - metabolism; Hippocampus - drug effects; Hippocampus - metabolism; L-Lactate Dehydrogenase - metabolism; Necrosis - drug therapy; Necrosis - metabolism; Nervous System Diseases - drug therapy; Nervous System Diseases - metabolism; Neurons - drug effects; Neurons - metabolism; Pharmacotherapy; Pharmacy; Rats; Rats, Sprague-Dawley; Short Communication; channel blockers; neurons; neuroprotection; Ca; glutamate; hippocampal culture
• ISSN: 1734-1140; 2299-5684
• DOI: 10.1016/S1734-1140(13)71052-1

Background
The aim of this study was to examine the putative protective effect of calcium channel blockers on hippocampal neurons in the experimental model of excitotoxic damage.
Methods
Seven-day old primary dissociated cultures of rat hippocampal neural cells containing one of the following calcium channel blockers: cinnarizine, flunarizine or nimodipine were exposed to glutamate-induced injury. Quantitative assessments of neuronal injury were accomplished by measuring lactate dehydrogenase (LDH) activity in the media 24 h after exposure to glutamate and by counting and establishing the apoptotic and necrotic cells in flow cytometry with Annexin V-FITC/PI staining.
Results
In our experiment, glutamate induced a 339% elevation of apoptotic cells and a 289% increase of necrotic cells in hippocampal neurons as compared to control cultures without drugs. In cultures containing flunarizine, glutamate-induced cell apoptosis was suppressed by 62% while necrosis showed no significant alternation. Cinnarizine exerted no anti-apoptotic effects on glutamate-injured cultured hippocampal neurons, while nimodipine intensified the apoptotic pathway of cell death and promoted an increase in the number of apoptotic neurons by 26%. When cinnarizine or nimodipine were used, the percentage of necrotic cells was significantly lower when compared with glutamate-injured cultures and it amounted to 44% and 24% for cinnarizine and nimodipine, respectively.
Conclusions
The obtained results suggest the beneficial anti-apoptotic potential of flunarizine and the anti-necrotic potential of cinnarizine against glutamate-induced death of cultured hippocampal neurons. Nimodipine can protect neurons against necrosis, but has an intensified adverse pro-apoptotic effect on cultured neurons in the experimental model of excitotoxic injury.