小剂量氯胺酮联用丁基苯酞的抗抑郁作用及其机制

Antidepressant Effects of Low-dose Ketamine Combination with n-Butylphthalide and Its Mechanism

目的研究小剂量氯胺酮(Ket)联用丁基苯酞(NBP)的抗抑郁作用及其机制。方法建立皮质酮(CORT)诱导PC12细胞损伤的体外抑郁模型,PC12细胞分成6组:空白对照组、模型对照组(CORT 400μmol·L^(-1))、对照A组(CORT 400μmol·L^(-1)+NBP 1μmol·L^(-1))、对照B组(CORT 400μmol·L^(-1)+Ket 0.01μmol·L^(-1))、阳性对照组(CORT 400μmol·L^(-1)+Ket 0.1μmol·L^(-1))、联合组(CORT 400μmol·L^(-1)+Ket 0.01μmol·L^(-1)+NBP 1μmol·L^(-1))。CCK-8法检测细胞存活率,荧光显微镜观察细胞神经元形态变化,蛋白免疫印迹法评价联合用药对损伤细胞p-ERK/ERK、脑源性神经营养因子(BDNF)、synapsin蛋白表达变化。将ICR小鼠随机分为模型对照组、NBP组(30 mg·kg^(-1))、Ket组(1 mg·kg^(-1))、联合组(NBP 30 mg·kg^(-1)+Ket 1 mg·kg^(-1))。每组分别给药,30 min后采用强迫游泳实验评价小鼠行为学变化。结果模型对照组、联合组细胞存活率分别为(52.17±4.56)%,(75.39±4.64)%,突触长度分别为(64.53±12.14),(89.12±24.53)μm,与模型对照组比较,联合组细胞存活率显著提高,神经元长度明显增长(均P<0.01);损伤细胞中p-ERK/ERK、BDNF蛋白表达水平显著提高(均P<0.05),且能够增加synapsin蛋白表达,但差异无统计学意义(P>0.05)。在小鼠急性抑郁模型实验中,模型对照组、联合组累计不动时间分别为(177.6±17.33),(119.90±41.45)s(P<0.01)。结论NBP能够增强小剂量Ket抗抑郁作用,且其作用机制可能与ERK-BDNF信号通路相关。

Objective To study antidepressant effects of low dose ketamine(Ket)combined with butylphthalide(NBP)and its mechanism.Methods In vitro depression model of PC12 cell injury by corticosterone(CORT)was established.PC12 cell were divided into six groups,blank control grup,model control group(CORT 400μmol·L^(-1)),control group A(CORT 400μmol·L^(-1)+NBP 1μmol·L^(-1)),control group B(CORT 400μmol·L^(-1)+Ket 0.01μmol·L^(-1)),positive group(CORT 400μmol·L^(-1)+Ket 0.1μmol·L^(-1)),combined group(CORT 400μmol·L^(-1)+Ket 0.01μmol·L^(-1)+NBP 1μmol·L^(-1)).Cell viability was detected by a CCK-8 method.The morphological changes of neurons were observed by a fluorescence microscopy.Western blotting was used to evaluate the expression levels of p-ERK/ERK,BDNF and synapsin in injured cells.ICR mice were randomly divided into model control group,NBP group(30 mg·kg^(-1)),Ket group(1 mg·kg^(-1))and combined group(NBP 30 mg·kg^(-1)+Ket 1 mg·kg^(-1)).Each group was administered separately.After 30 min from dosing,forced swimming test was used to evaluate the behavioral changes of mice.Results Compared with model control group,cell viabilities of model control group and combined group were(52.17±4.56)% and(75.39±4.64)%,synaptic neurons lengths were(64.53±12.14)μm and(89.12±24.53)μm,respectively.The neuronal length was significantly increased(all P<0.01),and the protein expression levels of p-ERK/ERK and BDNF in injured cells were significantly increased(all P<0.05),and the expression trend of synapsin protein was increased,but there was no statistical difference(P>0.05).In the acute depression mouse model experiment,the cumulative immobility times of the model control group and the combined group were(177.6±17.33)s and(119.90±41.45)s,respectively.Compared with the model control group,the cumulative immobility time of the combined group was significantly decreased(P<0.01).Conclusion NBP can enhance the antidepressant effect of low dose Ket,and its mechanism may be related to ERK-BDNF signaling pathway.