Herzfeld Eva, Speh Lea, Strauss Christian, Scheller Christian
Department of Neurosurgery, Martin-Luther University of Halle-Wittenberg, Ernst-Grube-Str. 40, 06120 Halle (Saale), Germany.
Int J Mol Sci. 2017 May 3;18(5):964. doi: 10.3390/ijms18050964.
Nimodipine is well characterized for the management of aneurysmal subarachnoid hemorrhage and has been shown to promote a better outcome and less delayed ischemic neurological deficits. Animal and clinical trials show neuroprotective efficacy following nerve injuries. We showed a neuroprotective effect on Neuro2a cells. Subsequent microarray analysis revealed-among others-fatty acid 2-hydroxylase (FA2H) upregulated by nimodipine in vitro, which is a component of myelin synthesis. Differentiated Neuro2a cells were analyzed for nimodipine-mediated survival considering stress treatment in comparison to nifedipine-treatment. Cell survival was determined by measurement of LDH activity in the culture medium. Nimodipine decreased surgery-like stress-induced cell death of differentiated Neuro2a cells. Neuro2a cell culture was analyzed for changes in FA2H expression induced by nimodipine or nifedipine in surgery-like stress conditions. We analyzed expression levels of FA2H mRNA and protein by qPCR using specific primers or a FA2H-specific antibody in nimodipine or nifedipine non- and pre-treated Neuro2a cell culture, respectively. Nimodipine but not nifedipine increases FA2H protein levels and also significantly increases mRNA levels of FA2H in both undifferentiated and differentiated Neuro2a cells. Our findings indicate that higher expression of FA2H induced by nimodipine may cause higher survival of Neuro2a cells stressed with surgery-like stressors.
尼莫地平在动脉瘤性蛛网膜下腔出血的治疗方面已得到充分研究，并且已证明其能促进更好的预后并减少延迟性缺血性神经功能缺损。动物和临床试验表明，神经损伤后尼莫地平具有神经保护作用。我们发现尼莫地平对Neuro2a细胞具有神经保护作用。随后的微阵列分析显示，在体外，尼莫地平上调了脂肪酸2 - 羟化酶（FA2H）等基因，FA2H是髓鞘合成的一个组成部分。为了研究尼莫地平介导的细胞存活情况，将分化的Neuro2a细胞在应激处理下与硝苯地平处理进行比较。通过测量培养基中的乳酸脱氢酶（LDH）活性来确定细胞存活率。尼莫地平减少了手术样应激诱导的分化Neuro2a细胞的死亡。在手术样应激条件下，分析了尼莫地平或硝苯地平诱导的Neuro2a细胞培养物中FA2H表达的变化。我们分别在未用和预先用尼莫地平或硝苯地平处理的Neuro2a细胞培养物中，使用特异性引物通过定量聚合酶链反应（qPCR）分析FA2H mRNA水平，或使用FA2H特异性抗体分析FA2H蛋白表达水平。尼莫地平而非硝苯地平可增加FA2H蛋白水平，并且在未分化和分化的Neuro2a细胞中均显著增加FA2H的mRNA水平。我们的研究结果表明，尼莫地平诱导的FA2H高表达可能导致在手术样应激源作用下的Neuro2a细胞具有更高的存活率。
