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目的 探讨氯胺酮对体外培养的吗啡慢性处理脊髓星形胶质细胞释放谷氨酸的影响。方法体外培养星形胶质细胞于融合状态,随机分为8组:对照组(不加吗啡)和吗啡依赖模型A、B1、B2、B3、C1、C2、C3组,48h后各组培养液均换成Neumbasal/B27无血清培养液,A组不加任何药物,B1、B2、B3组分别加入终浓度为0.1、1、10μmol·L-1的吗啡,C1、C2、C3组分别加入0.4、4、40μmol·L-1的氯胺酮,后6组加药15 min后再加入终浓度为10μmol·L-1纳络酮。用反相高效液相色潜分析法测定各组细胞外液中谷氨酸含量。结果 对照组谷氨酸含量[(2.52±0.33)μmol·L-1]与A组[(2.61±0.27)μmol·L-1]比较差异无显著性(P>0.05)。B1、B2、B3组内谷氨酸的含量均升高,其中B1组谷氨酸含量[(6.36±2.19)μmol·L-1]与A组[(2.61±0.27)μmol·L-1]相比差异有非常显著性(P<0.01).B2、B3组与A组相比,差异有显著性(P<0.05)。C1、C2、C3组内谷氨酸含量则随着氯胺酮预处理量的增加而逐渐降低。结论 星形胶质细胞可能通过调节胞外谷氨酸浓度的变化来参与吗啡依赖和戒断反应,而氯胺酮则通过减少星形胶质细胞释放谷氨酸来发挥抗戒断症状的作用。
Objective To investigate the effects of ketamine on glutamate release from chronic morphine treated spinal cord astrocytes in vitro. Methods The astrocytes cultured in vitro were randomly divided into 8 groups: control group (without morphine) and morphine-dependent models A, B1, B2, B3, C1, C2, Nembasal / B27 serum-free medium was replaced with no drug in group A, morphine was added in groups B1, B2 and B3 respectively, and groups C1, C2 and C3 were added with 0.4 , 4, 40μmol·L-1 ketamine. After the last six groups were added with drugs for 15 min, naloxone at a final concentration of 10μmol·L-1 was added. The contents of glutamic acid in extracellular fluid of each group were determined by RP-HPLC. Results There was no significant difference in glutamate content in the control group [(2.52 ± 0.33) μmol·L-1 vs [2.61 ± 0.27] μmol·L-1] in the control group (P> 0.05). The contents of glutamic acid in group B1, B2 and B3 were all higher than those in group A [(6.36 ± 2.19) μmol·L-1 in group B1 [(2.61 ± 0.27) μmol·L-1] (P <0.01), there was a significant difference between group B2 and group B3 (P <0.05). The contents of glutamic acid in groups C1, C2 and C3 decreased with the pretreatment of ketamine. Conclusion Astrocytes may participate in the morphine dependence and withdrawal response by regulating the concentration of extracellular glutamate, while ketamine exerts anti-withdrawal symptoms by reducing glutamate release from astrocytes.