Prolonged Treatment with Propofol Transiently Impairs Proliferation but Not Survival of Rat Neural Progenitor Cells In Vitro.
Prolonged Treatment with Propofol Transiently Impairs Proliferation but Not Survival of Rat Neural Progenitor Cells In Vitro.
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DOI：
10.1371/journal.pone.0158058
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Culley DJ
中科院分区：
文献类型：
--
作者：
Palanisamy A;Friese MB;Cotran E;Moller L;Boyd JD;Crosby G;Culley DJ
神经认知功能障碍在重症监护幸存者中很常见。这与长时间的镇静有关，但其机制尚不清楚。神经发生持续到成年期，并与学习有关。驱动神经发生的神经前体细胞(NPC)具有临床上使用的主要镇静剂的受体，这表明神经发生的中断可能在一定程度上导致ICU幸存者的认知能力下降。使用体外系统，我们测试了假设，长期暴露于异丙酚浓度-和持续时间-依赖于杀死或显著减少鼻咽癌细胞的增殖。从胚胎第14天的SD大鼠幼鼠分离的鼻咽癌细胞分别暴露于0、2.5或5.0μg/mL异丙酚，其浓度与深度临床麻醉一致，持续4或24小时。在丙泊酚暴露后立即或24小时后检测细胞的死亡和增殖情况。分别用丙啶碘染色和裂解caspase-3免疫细胞化学法检测鼻咽癌细胞的死亡和凋亡率，用EDU掺入法检测鼻咽癌细胞的增殖。用星形孢子素(1μM作用6h)作为阳性对照。用无偏高通量免疫细胞化学方法分析细胞。无论是丙泊酚浓度还是暴露时间，都没有细胞死亡。丙泊酚作用4h对鼻咽癌细胞增殖无明显影响，但作用24 h后，两种浓度的异丙酚均有抑制鼻咽癌细胞增殖的作用(P<；0.0001，与对照组比较)。然而，这种作用是短暂的，停用异丙酚24小时后，细胞增殖恢复到基线水平(P=0.37，异丙酚与对照组相比)。异丙酚暂时但可逆地抑制了鼻咽癌的增殖，没有细胞毒性，对神经干细胞池的影响可以忽略不计，这表明异丙酚即使在临床麻醉中使用的浓度，对神经前体细胞生物学的影响也是有限的。
Neurocognitive dysfunction is common in survivors of intensive care. Prolonged sedation has been implicated but the mechanisms are unclear. Neurogenesis continues into adulthood and is implicated in learning. The neural progenitor cells (NPC) that drive neurogenesis have receptors for the major classes of sedatives used clinically, suggesting that interruption of neurogenesis may partly contribute to cognitive decline in ICU survivors. Using an in vitro system, we tested the hypothesis that prolonged exposure to propofol concentration- and duration-dependently kills or markedly decreases the proliferation of NPCs. NPCs isolated from embryonic day 14 Sprague-Dawley rat pups were exposed to 0, 2.5, or 5.0 μg/mL of propofol, concentrations consistent with deep clinical anesthesia, for either 4 or 24 hours. Cells were assayed for cell death and proliferation either immediately following propofol exposure or 24 hours later. NPC death and apoptosis were measured by propidium iodine staining and cleaved caspase-3 immunocytochemistry, respectively, while proliferation was measured by EdU incorporation. Staurosporine (1μM for 6h) was used as a positive control for cell death. Cells were analyzed with unbiased high-throughput immunocytochemistry. There was no cell death at either concentration of propofol or duration of exposure. Neither concentration of propofol impaired NPC proliferation when exposure lasted 4 h, but when exposure lasted 24 h, propofol had an anti-proliferative effect at both concentrations (P < 0.0001, propofol vs. control). However, this effect was transient; proliferation returned to baseline 24 h after discontinuation of propofol (P = 0.37, propofol vs. control). The transient but reversible suppression of NPC proliferation, absence of cytotoxicity, and negligible effect on the neural stem cell pool pool suggest that propofol, even in concentrations used for clinical anesthesia, has limited impact on neural progenitor cell biology.