Effects of Sevoflurane on Self-Renewal Capacity and Differentiation of Cultured Neural Stem Cells
Effects of Sevoflurane on Self-Renewal Capacity and Differentiation of Cultured Neural Stem Cells
复制标题
七氟醚对培养神经干细胞自我更新能力和分化的影响
DOI：
10.1007/s11064-013-1074-4
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发表时间：
2013-08-01
影响因子：
4.4
通讯作者：
Xiong, Lize
中科院分区：
文献类型：
--
作者：
Nie, Huang;Peng, Zhengwu;Xiong, Lize
七氟醚麻醉可导致幼鼠长期认知障碍，可能是通过抑制神经发生。海马体对记忆巩固至关重要，是哺乳动物大脑中仅有的两个神经干细胞（NSCs）在一生中不断更新的区域之一。为了阐明七氟醚诱导认知功能障碍的发病机制，我们测量了临床七氟醚剂量对海马NSCs存活、增殖和分化的影响。从Sprague-Dawley大鼠胚胎中分离神经干细胞，体外扩增，并以0.5、1或1.5的最低肺泡浓度（MAC）暴露于七氟醚中1或6小时。处理后2天，分别通过WST-1测定、乳酸脱氢酶（LDH）活性和tdt介导的dutp -生物素缺口末端标记（TUNEL）来评估细胞活力、细胞毒性和凋亡率。5-乙基-2′-脱氧尿苷（BrdU）掺入及Ki67染色检测细胞增殖率。用免疫细胞化学和Western blot检测治疗后7天的分化情况。在增殖期和分化期分别测定p44/42细胞外调节激酶（ERK1/2）的磷酸化水平。七氟醚在1 MAC或1.5 MAC下作用1 h可增加活细胞数量，而在相同浓度下作用6 h可抑制细胞增殖并促进细胞凋亡（P < 0.01）。七氟醚对NSC分化没有影响，亚临床浓度（0.5 MAC）既不改变增殖也不改变活力。七氟醚暴露1 MAC或1.5 MAC后1 h， ERK1/2的磷酸化水平在增殖期升高，而在分化期没有升高。短暂（1小时）暴露于临床浓度的七氟醚可增强培养的NSCs的增殖，这可能是由ERK1/2介导的，但暴露6小时可抑制增殖并诱导凋亡。长时间暴露于七氟醚可降低海马NSCs的自我更新能力，导致认知缺陷。
Sevoflurane anesthesia in infant rats can result in long-term cognitive impairment, possibly by inhibiting neurogenesis. The hippocampus is critical for memory consolidation and is one of only two mammalian brain regions where neural stem cells (NSCs) are renewed continuously throughout life. To elucidate the pathogenesis of sevoflurane-induced cognitive dysfunction, we measured the effects of clinical sevoflurane doses on the survival, proliferation, and differentiation of hippocampal NSCs. Neural stem cells were isolated from Sprague-Dawley rat embryos, expanded in vitro, and exposed to sevoflurane at 0.5, 1, or 1.5 minimal alveolar concentration (MAC) for 1 or 6 h. Two days after treatment, cell viability, cytotoxicity, and apoptosis rate were estimated by WST-1 assay, lactate dehydrogenase (LDH) activity, and TdT-mediated dUTP-biotin nick end labeling (TUNEL), respectively, while proliferation rate was assessed by 5-ethynyl-2'-deoxyuridine (BrdU) incorporation and Ki67 staining. Differentiation was assayed 7 days after treatment by immunocytochemistry and Western blots of neuron and glial markers. The phosphorylation level of p44/42 extracellular regulated kinases (ERK1/2) was measured in the proliferation and differentiation phases respectively. Sevoflurane at 1 MAC or 1.5 MAC for 1 h increased viable cell number whereas a 6 h exposure at these same concentrations suppressed proliferation and promoted apoptotic death (P < 0.01). Sevoflurane had no effect on NSC differentiation, and a sub-clinical concentration (0.5 MAC) altered neither proliferation nor viability. The phosphorylation level of ERK1/2 increased after 1 h of 1 MAC or 1.5 MAC of sevoflurane exposure in the proliferation phase, but not in the differentiation phase. Brief (1 h) exposure to sevoflurane at clinical concentrations enhanced proliferation of cultured NSCs possibly mediated by ERK1/2, but a 6 h exposure suppressed proliferation and induced apoptosis. Prolonged sevoflurane exposure may decrease the self-renewal capacity of hippocampal NSCs, resulting in cognitive deficits.