氯胺酮复合咪达唑仑对谷氨酸诱导PC12细胞凋亡的影响

Effects of ketamine combined with midazolam on glutamate-induced apoptosis in PC12 cells

目的探讨氯胺酮复合咪达唑仑对谷氨酸诱导PCI2细胞凋亡的影响。方法诱导分化4d的神经元样PCI2细胞随机分为5组：对照组（C组）；谷氨酸组（Glu组）加入20mmol／L谷氨酸；氯胺酮组（K组）、咪达唑仑组（M组）、氯胺酮＋咪达唑仑组（K＋M组）均加入20mmol／L谷氨酸后，分别加入50μmol／L氯胺酮、1μmol／L咪达唑仑、50μmol／L氯胺酮＋1μmol／L咪达唑仑。各组细胞继续培养24h后采用M，IT法检测细胞活力，Hoechst33258核染色法及Annexin V—FITC／PI双染流式细胞术检测凋亡细胞。结果与C组比较，Glu组细胞活力降低，细胞凋亡率升高（P〈0．01）；与Glu组比较，K组和M组细胞活力升高，细胞凋亡率降低（P〈0．05）；与K组和M组比较，K＋M组细胞活力升高，细胞凋亡率降低（P〈0．05）；K组和M组细胞活力及细胞凋亡率差异无统计学意义（P〉0．05）。结论氯胺酮复合咪达唑仑可更有效地抑制谷氨酸诱导PCI2细胞凋亡。

Objective To investigate the effects of ketamine combined with midazolam on glutamateinduced apoptosis in neuronal PC12 cells. Methods PC12 cells were provided by animal experiment center, SUN Yat-Sen University. After being cultured in DMEM liguid culture medium containing 2.5 S-NGF （nerve growth factor） in an incubator filled with 5% CO2-95% 02 at 37℃ for 4 days, over 90% of the PC12 cells were differentiated into neuron-type cells. The cells were then randomly divided into 5 groups： control group （group C） ; glutamate group （group Glu）, the cells were incubated in the presence of glutamate 20 mmol/L for 24 h; ketamine group （group K）; midazolam group （group M） and ketamine-midazolam group （group K ＋ M）, the cells were incubated with glutamate 20 mmol/L ＋ ketamine 50 μmol/L or midazolam 1 μmol/L or ketamine 50 μmol/L ＋ midazolam 1 μmol/L for 24 h. The cell viability was evaluated by MTT assay. The apoptosis was detected by Hoechst 33258 nucleus staining and flow cytometry with Annexin V-FITC/PI staining. Results The viability of PC12 cells was significantly decreased while the apoptosis rate was significantly increased by incubation with glutamate 20 mmol/L for 24 h as compared with control group. Ketamine and midazolam alone or in combination attenuated the glutamate-induced injury to the cells. Conclusion Ketamine combined with midazolam can effectively protect against glutamate-induced apoptosis in neuronal PC12 cells.