氯胺酮复合咪达唑仑对谷氨酸诱导PC12细胞凋亡的影响

目的 探讨氯胺酮复合咪达唑仑对谷氨酸诱导PC12细胞凋亡的影响.方法 诱导分化4 d的神经元样PC12细胞随机分为5组:对照组(c组);谷氨酸组(Glu组)加入20 mmol/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20 mmol/L谷氨酸后,分别加入50 μmol/L氯胺酮、1μmol/L咪达唑仑、50 μmol/L氯胺酮+1 μmol/L咪达唑仑.各组细胞继续培养24 h后采用MTT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PC12细胞凋亡.

Effects of ketamine combined with midazolam on glutamate-induced apoptosis in PCI2 cells

Objective To investigate the effects of ketamine combined with midazolam on glutamate-induced apoptosis in neuronal PC12 cells.Methods PCI2 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% CO_2-95% O_2 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 + ketaminc 50 μmol/L or midazolam 1 μmol/L or ketamine 50 μmol/L + midazolam I 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 midazolum 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.