Ketamine alters the neurogenesis of rat cortical neural stem progenitor cells.
Ketamine alters the neurogenesis of rat cortical neural stem progenitor cells.
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DOI：
10.1097/ccm.0b013e318253563c
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发表时间：
2012-08
影响因子：
8.8
通讯作者：
Anand KJ
中科院分区：
文献类型：
--
作者：
Dong C;Rovnaghi CR;Anand KJ
高剂量或长时间暴露于氯胺酮会增加发育中大脑的神经元凋亡，尽管对神经干祖细胞（NSPCs）的影响尚未研究。本研究探讨了氯胺酮对培养的大鼠胎脑皮层神经干细胞的细胞死亡和神经发生的剂量和时间依赖性反应。基于实验室的研究大学研究实验室Sprague-Dawley（SD）大鼠NSPCs从胚胎第17天（E17）的SD大鼠胎儿的皮质中分离。在剂量反应实验中，将培养的NSPCs暴露于不同浓度的氯胺酮（0-100 μM）24小时。在时程实验中，将NSPC培养物暴露于10 μM氯胺酮不同持续时间（0-48小时）。分别用活化的caspase-3免疫染色和乳酸脱氢酶（LDH）测定评估NSPCs的凋亡和坏死。用溴脱氧尿苷（BrdU）掺入法和Ki 67免疫组化法检测NSPCs的增殖变化。使用Tuj-1免疫染色评估神经元分化。在所有浓度和持续时间的氯胺酮暴露试验后，培养的NSPCs对凋亡和坏死具有抗性。氯胺酮可抑制增殖，氯胺酮暴露于100 μM 24小时后BrdU阳性细胞数量减少在氯胺酮浓度高于10 μM（P <0.005）或10 μM（P<0.01）48小时后，Ki 67阳性细胞的数量减少（P<0.001），而在氯胺酮浓度高于10 μM（P<0.001）或10 μM（P<0.01）48小时后，Ki 67阳性细胞的数量减少（P<0.01）。氯胺酮增强神经元分化，暴露24小时后，所有氯胺酮浓度均增加Tuj-1阳性神经元（P<0.001）。所有暴露于10 μM氯胺酮超过8小时的情况也发生了这种情况（P<0.001）。临床相关浓度的氯胺酮不会通过细胞凋亡或坏死诱导NSPC中的细胞死亡。氯胺酮改变了从大鼠新皮层分离的NSPCs的增殖并增加了其神经元分化。这些研究表明，在胎儿或新生儿生命中暴露于氯胺酮可能会改变神经发生和随后的大脑发育。
High doses or prolonged exposure to ketamine increase neuronal apoptosis in the developing brain, although effects on neural stem progenitor cells (NSPCs) remain unexplored. This study investigated dose- and time- dependent responses to ketamine on cell death and neurogenesis in cultured rat fetal cortical NSPCs. Laboratory-based study University research laboratory Sprague-Dawley (SD) rats NSPCs were isolated from the cortex of SD rat fetuses on embryonic day 17 (E17). In dose-response experiments, cultured NSPCs were exposed to different concentrations of ketamine (0–100 μM) for 24 hours. In time-course experiments, NSPC cultures were exposed to 10 μM ketamine for different durations (0–48 hours). Apoptosis and necrosis in NSPCs were assessed using activated caspase-3 immunostaining and lactate dehydrogenase (LDH) assays, respectively. Proliferative changes in NSPCs were detected using Bromo-deoxyuridine (BrdU) incorporation and Ki67 immunostaining. Neuronal differentiation was assessed using Tuj-1 immunostaining. Cultured NSPCs were resistant to apoptosis and necrosis following all concentrations and durations of ketamine exposure tested. Ketamine inhibited proliferation, with decreased numbers of BrdU-positive cells following ketamine exposure to 100 μM for 24 hours (P<0.005) or 10 μM for 48 hours (P<0.01), and reduced numbers of Ki67-positive cells following exposure to ketamine concentration higher than 10 μM for 24 hours (P<0.001) or at 10 μM for 48 hours (P<0.01). Ketamine enhanced neuronal differentiation, with all ketamine concentrations increasing Tuj-1-positive neurons (P<0.001) after 24-hours of exposure. This also occurred with all exposures to 10 μM ketamine for longer than 8 hours (P<0.001). Clinically relevant concentrations of ketamine do not induce cell death in NSPCs via apoptosis or necrosis. Ketamine alters the proliferation and increases the neuronal differentiation of NSPCs isolated from the rat neocortex. These studies imply that ketamine exposure during fetal or neonatal life may alter neurogenesis and subsequent brain development.