Zagaja Mirosław, Haratym-Maj Agnieszka, Szewczyk Aleksandra, Rola Radoslaw, Maj Maciej, Łuszczki Jarogniew J, Andres-Mach Marta
Isobolographic Analysis Laboratory, Institute of Rural Health, Jaczewskiego 2, 20-090 Lublin, Poland.
Department of Physiopathology, Institute of Rural Health, Jaczewskiego 2, 20-090 Lublin, Poland.
Neurosci Lett. 2019 Mar 23;696:79-86. doi: 10.1016/j.neulet.2018.12.016. Epub 2018 Dec 12.
The aim of the study was to evaluate the impact of second generation antiepileptic drug levetiracetam (LEV) with arachidonyl-2'-chloroethylamide (ACEA) on proliferating neural precursor cells in mouse brain. Additionally, we established the relationship between treatment with ACEA in combination with LEV and hippocampal neurogenesis in mouse brain. All experiments were performed on male CB57/BL mice injected i.p. with LEV (10 mg/kg), ACEA (10 mg/kg) and PMSF (30 mg/kg) for 10 days. Experiments were provided in two stages: stage 1- an acute response of proliferating neural precursor cells to ACEA and LEV administration (Ki-67 staining), stage 2 - a long term response to ACEA and LEV administration (BrDU, NeuN, GFAP staining). Results indicate that ACEA + PMSF and ACEA + PMSF + LEV significantly increased the total number of Ki-67 positive cells comparing to the control group. PMSF and LEV administered alone and in combination had no significant impact on cell proliferation compared to the control group. Results from neurogenesis study indicated that ACEA + PMSF administered alone and in combination with LEV increased the total number of BrDU cells compared to the control group, although LEV on its own decreased the number of BrDU cells. Moreover, the combination of ACEA + PMSF + LEV significantly increased the total number of newborn neurons compared to the control group. In turn, LEV significantly decreased the process of neurogenesis. Astrocytes were considerably reduced in all treated groups as compare to the control mice. These data provide substantial evidence that LEV administered chronically decreases the proliferation and differentiation of newly born cells while combination of LEV + ACEA significantly increases the level of newborn neurons in the dentate subgranular zone.
本研究的目的是评估第二代抗癫痫药物左乙拉西坦（LEV）与花生四烯酸-2'-氯乙酰胺（ACEA）对小鼠脑内增殖神经前体细胞的影响。此外，我们还建立了ACEA与LEV联合治疗与小鼠脑海马神经发生之间的关系。所有实验均在腹腔注射LEV（10mg/kg）、ACEA（10mg/kg）和苯甲基磺酰氟（PMSF，30mg/kg）10天的雄性CB57/BL小鼠身上进行。实验分两个阶段进行：第一阶段——增殖神经前体细胞对ACEA和LEV给药的急性反应（Ki-67染色），第二阶段——对ACEA和LEV给药的长期反应（溴脱氧尿苷（BrDU）、神经元核抗原（NeuN）、胶质纤维酸性蛋白（GFAP）染色）。结果表明，与对照组相比，ACEA + PMSF以及ACEA + PMSF + LEV显著增加了Ki-67阳性细胞的总数。单独给药及联合给药的PMSF和LEV与对照组相比，对细胞增殖无显著影响。神经发生研究结果表明，与对照组相比，单独给予ACEA + PMSF以及与LEV联合给药均增加了BrDU细胞的总数，尽管单独使用LEV会减少BrDU细胞的数量。此外，与对照组相比，ACEA + PMSF + LEV联合用药显著增加了新生神经元的总数。相反，LEV显著降低了神经发生的过程。与对照小鼠相比，所有治疗组的星形胶质细胞均显著减少。这些数据提供了充分的证据，即长期给予LEV会降低新生细胞的增殖和分化，而LEV + ACEA联合用药则显著增加齿状回颗粒下区新生神经元的水平。
