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 ([Ca²⁺]i) elevation was monitored in cells on DIV 4, 8, and 13. All anesthetics elicited significant [Ca²⁺]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 [Ca²⁺]i elevation or SR decrease, while PPF still induced [Ca²⁺]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 [Ca²⁺]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.

麻醉剂引起的大脑发育中的神经毒性是一个令人关注的问题。这种神经毒性与麻醉剂的暴露时间，剂量和发育阶段密切相关。使用体外钙成像和形态学检查，我们试图确定静脉麻醉药诱导的直接神经毒性是否根据原代培养物中神经元体外培养天数（DIV）的不同阶段而变化。制备了来自E17 Wistar大鼠的皮质神经元。在DIV 3、7和13上，将细胞暴露于静脉麻醉药硫喷妥钠（TPS），咪达唑仑（MDZ）或丙泊酚（PPF），以使用形态学实验研究直接的神经毒性。此外，使用钙成像技术，麻醉剂诱导的细胞内钙浓度（[Ca ²⁺] i）在DIV 4、8和13上的细胞中进行了监测。所有麻醉剂在DIV 4上引起[Ca ²⁺ ] i显着增加。虽然TPS（100μM）和MDZ（10μM）不会改变神经元的死亡， PPF（10μM和100μM）显着降低了生存率（SR）。在DIV 8上，TPS和MDZ不会引起[Ca ²⁺ ] i升高或SR降低，而PPF仍会引起[Ca ²⁺ ] i升高（在10μM和100μM时），并且在100μM时SR显着降低（0.76） ±0.03; P <0.05），但不是在10μM（0.91±0.03）时。在任何麻醉药的DIV 13-14上均未观察到这种麻醉药诱导的[Ca ²⁺ ] i升高和SR降低。我们的研究表明，与TPS或MDZ相比，在开发过程中使用PPF可能要更加谨慎。