Department of Anesthesiology and Intensive Care Medicine, International University of Health and Welfare, Narita, Chiba, Japan; Department of Anesthesiology and Intensive Care Medicine, Osaka University, Suita, Osaka, Japan.
Department of Anesthesiology and Intensive Care Medicine, Osaka University, Suita, Osaka, Japan.
Neurotoxicology. 2018 Dec;69:320-329. doi: 10.1016/j.neuro.2018.08.002. Epub 2018 Aug 11.
Anesthetic-induced neurotoxicity in the developing brain is a concern. This neurotoxicity is closely related to anesthetic exposure time, dose, and developmental stages. Using calcium imaging and morphological examinations in vitro, we sought to determine whether intravenous anesthetic-induced direct neurotoxicity varies according to different stages of the days in vitro (DIV) of neurons in primary culture. Cortical neurons from E17 Wistar rats were prepared. On DIV 3, 7, and 13, cells were exposed to the intravenous anesthetics thiopental sodium (TPS), midazolam (MDZ), or propofol (PPF), to investigate direct neurotoxicity using morphological experiments. Furthermore, using calcium imaging, the anesthetic-induced intracellular calcium concentration ([Ca]i) elevation was monitored in cells on DIV 4, 8, and 13. All anesthetics elicited significant [Ca]i increases on DIV 4. While TPS (100 μM) and MDZ (10 μM) did not alter neuronal death, PPF (10 μM and 100 μM) decreased the survival ratio (SR) significantly. On DIV 8, TPS and MDZ did not elicit [Ca]i elevation or SR decrease, while PPF still induced [Ca]i elevation (both at 10 μM and 100 μM) and significant SR decrease at 100 μM (0.76 ± 0.03; P < 0.05), but not at 10 μM (0.91 ± 0.03). Such anesthetic-induced [Ca]i elevation and SR decrease were not observed on DIV 13-14 for any of the anesthetic drugs. Our study indicates that more caution may be exercised when using PPF compared to TPS or MDZ during development.
在发育中的大脑中，麻醉诱导的神经毒性是一个令人关注的问题。这种神经毒性与麻醉暴露时间、剂量和发育阶段密切相关。我们通过体外钙成像和形态学检查，试图确定静脉麻醉诱导的直接神经毒性是否因原代培养神经元的体外日龄（DIV）不同而有所不同。从 E17 Wistar 大鼠中制备皮质神经元。在 DIV3、7 和 13 时，用静脉麻醉剂硫喷妥钠（TPS）、咪达唑仑（MDZ）或丙泊酚（PPF）处理细胞，通过形态学实验研究直接神经毒性。此外，在 DIV4、8 和 13 时，通过钙成像监测麻醉剂诱导的细胞内钙浓度（[Ca]i）升高。所有麻醉剂在 DIV4 时均引起显著的[Ca]i 升高。虽然 TPS（100 μM）和 MDZ（10 μM）不会改变神经元死亡，但 PPF（10 μM 和 100 μM）显著降低了存活率（SR）。在 DIV8 时，TPS 和 MDZ 不会引起[Ca]i 升高或 SR 降低，而 PPF 仍会引起[Ca]i 升高（在 10 μM 和 100 μM 时），并且在 100 μM 时（0.76 ± 0.03；P < 0.05），但在 10 μM 时（0.91 ± 0.03）则不会显著降低 SR。在任何麻醉剂中，在 DIV13-14 时都没有观察到这种麻醉诱导的[Ca]i 升高和 SR 降低。我们的研究表明，在发育过程中使用 PPF 时可能需要更加谨慎，与 TPS 或 MDZ 相比。
