钙通道阻滞剂对谷氨酸诱导培养海马神经元损伤的保护作用研究。

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

机构信息

Department of Pediatric Neurology and Rehabilitation, Medical University of Bialystok, Białystok, Poland.

出版信息

Pharmacol Rep. 2013;65(3):730-6. doi: 10.1016/s1734-1140(13)71052-1.

DOI:10.1016/s1734-1140(13)71052-1

PMID:23950597

Abstract

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.

摘要

背景

本研究旨在探讨钙通道阻滞剂在兴奋性毒性损伤的实验模型中对海马神经元的潜在保护作用。

方法

将含有钙通道阻滞剂肉桂嗪、氟桂利嗪或尼莫地平之一的 7 天龄大鼠海马神经细胞原代分离培养物暴露于谷氨酸诱导的损伤中。通过测量暴露于谷氨酸 24 小时后培养基中乳酸脱氢酶（LDH）的活性以及通过 Annexin V-FITC/PI 染色在流式细胞术中计数和确定凋亡和坏死细胞，对神经元损伤进行定量评估。

结果

在我们的实验中，与不含药物的对照培养物相比，谷氨酸诱导海马神经元中凋亡细胞增加 339%，坏死细胞增加 289%。在含有氟桂利嗪的培养物中，谷氨酸诱导的细胞凋亡抑制了 62%，而坏死没有明显改变。肉桂嗪对谷氨酸损伤的培养海马神经元没有抗凋亡作用，而尼莫地平加剧了细胞死亡的凋亡途径，并使凋亡神经元的数量增加了 26%。当使用肉桂嗪或尼莫地平时，与谷氨酸损伤培养物相比，坏死细胞的百分比显著降低，肉桂嗪和尼莫地平分别为 44%和 24%。