Zhao Yilin, Jin Xiaogao, Wang Jintao, Tan Lei, Li Shiyong, Luo Ailin
Department of Anesthesiology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, China.
J Huazhong Univ Sci Technolog Med Sci. 2011 Dec;31(6):779-783. doi: 10.1007/s11596-011-0676-9. Epub 2011 Dec 16.
This study examined the effects of clinically relevant concentrations of isoflurane on the amplitude of NMDA receptor current (I(NMDA)) and the expression of cytochrome C in cultured developing rat hippocampal neurons. The hippocampi were dissected from newborn Sprague-Dawley rats. Hippocampal neurons were primarily cultured for 5 days and then treated with different concentrations of isoflurane [(0.25, 0.5, 0.75, 1 minimum alveolar concentration (MAC))]. The peak of I(NMDA) was recorded by means of the whole cell patch clamp technique. The cytochrome C level was detected by Western blotting and quantitative real-time PCR. Our results showed that isoflurane (0.25, 0.5, 0.75 and 1 MAC) potentiated the amplitude of I(NMDA) by (116 ± 8.8)%, (122 ± 11.7)%, (135 ± 14.3)% and (132 ± 14.6)%, respectively, and isoflurane increased the mRNA expression of cytochrome C in a concentration-dependent manner. The cytochrome C mRNA expression reached a maximum after 0.5 MAC isoflurane stimulation for 6 h (P<0.05). It was concluded that isoflurane enhances the expression of cytochrome C in cultured rat hippocampal neurons, which may be mediated by facilitation of NMDA receptor.
本研究检测了临床相关浓度的异氟烷对培养的发育中大鼠海马神经元N-甲基-D-天冬氨酸受体电流(I(NMDA))幅度及细胞色素C表达的影响。从新生的Sprague-Dawley大鼠中分离出海马体。将海马神经元原代培养5天,然后用不同浓度的异氟烷[(0.25、0.5、0.75、1最低肺泡浓度(MAC))]进行处理。采用全细胞膜片钳技术记录I(NMDA)的峰值。通过蛋白质免疫印迹法和定量实时聚合酶链反应检测细胞色素C水平。我们的结果显示,异氟烷(0.25、0.5、0.75和1 MAC)分别使I(NMDA)的幅度增强了(116±8.8)%、(122±11.7)%、(135±14.3)%和(132±14.6)%,并且异氟烷以浓度依赖的方式增加细胞色素C的mRNA表达。在0.5 MAC异氟烷刺激6小时后,细胞色素C的mRNA表达达到最大值(P<0.05)。得出的结论是,异氟烷增强了培养的大鼠海马神经元中细胞色素C的表达,这可能是由N-甲基-D-天冬氨酸受体的促进作用介导的。