Nimodipine but Not Nifedipine Promotes Expression of Fatty Acid 2-Hydroxylase in a Surgical Stress Model Based on Neuro2a Cells

Abstract

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 fa2h 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.

Nimodipine-Dependent Protection of Schwann Cells, Astrocytes and Neuronal Cells from Osmotic, Oxidative and Heat Stress Is Associated with the Activation of AKT and CREB

“…In previous studies, nimodipine concentrations between 1–20 µM were appropriate for achieving a neuroprotective effect in cell culture [11,12], which also applies to the present results. This suggests that the neuroprotective effects of nimodipine are not dose-dependent in the concentrations between 1–20 µM, but that the time of treatment seems to be critical.…”

“…Our data have shown that there is a neuroprotective effect as a result of a 24-h pretreatment with nimodipine. The stress models were established in previous studies [11,12,26]. Ethanol treatment is able to induce oxidative stress through acetaldehyde metabolism, which inhibits mitochondrial respiration at the level of complex I (NADH-ubiquinone oxido-reductase) and the coupling site I of oxidative phosphorylation [27].…”

“…As a voltage-dependent L-type calcium channel blocker, nimodipine regulates the calcium influx in the cells [56,57], which could prevent calcium overload causing apoptosis. In contrast, nifedipine, which is also a calcium channel blocker of the same class and which has a high structural identity with nimodipine, showed no or lower neuroprotective properties [12,58]. Therefore, further studies should clarify whether the shown anti-apoptotic mechanism is controlled by the prevention of calcium overload.…”

“…Cytotoxicity was induced with 2% EtOH, 150 mM NaCl and a 6-h heat incubation at 42 °C, as described in Herzfeld et al 2017 [12] and shown schematically in Figure 7.…”

“…Neuroprotection through nimodipine pretreatment has been demonstrated in vitro in PC-12 pheochromocytoma cells [10]. We have shown in previous studies that the cell death of neuroblastoma cells was significantly reduced after pretreatment with nimodipine prior to stress induction [11,12]. In addition, an upregulation of fatty acid 2-hydrolase (FA2H) was shown via nimodipine and stress induction on the mRNA and protein levels.…”