| Objective:To observe the protective effect of ketamine?Ket?combined with butylphathlide?NBP?on PC12 cell injury induced by corticosterone?CORT?and the effect of the combination of ketamine and NBP on acute depression like behavior,and to explore the molecular mechanism of the combination of ketamine and NBP in the development and application of antidepressants.Methods:The model of PC12 cell injury induced by corticosterone was established.The protective effect of ketamine and butylphthalide on PC12 cell injury induced by corticosterone was detected by CCK-8 method,and the changes of synaptic morphology of PC12 cell under fluorescence microscope were observed.The model of acute depression was used to observe the effect of ketamine combined with butylphthalide on the behavior of depression like mice.Western blot was used to detect the effect of ketamine combined with butylphthalide on the expression of PC12 cell and p-ERK,BDNF,p-mTOR,synapsin and other related proteins in the cerebral cortex of depression like mice.Results:In the cell experiment,corticosterone showed a dose-dependent effect on PC12 cell injury and significantly reduced the cell survival rate at 400?M?P<0.001?.Compared with the model group,ketamine group?Ket,0.01?M?and butylphthalide group?NBP,1?M?had no difference,while the combination group?0.01?M Ket+1?M NBP?and Ket group?0.1?M?could significantly improve the survival rate of injured cells?P<0.01,P<0.01?.The results of cell synapse morphology showed that compared with the control group,the axon length of cortisone group was significantly shorter?P<0.001?.The axon length of the injured cells increased significantly in the combination group and 0.1?m Ket group?P<0.01,P<0.01?.Animal behavior experiments found that compared with the solvent control group,there was no significant difference in the cumulative immobility time of acute depression like mice treated with ketamine(1mg·kg-1)or butylphthalide(30mg·kg-1),but the combination of ketamine and butylphthalide could significantly reduce the cumulative immobility time of forced swimming?P<0.01?,and ketamine 15mg·kg-1 also significantly reduced the accumulated immobility time in forced swimming?P<0.001?.In the open field experiment,the results showed that 1 mg·kg-1 ketamine group,30 mg·kg-1butylphthalide group and the combination group had no effect on the number of horizontal climbing and standing of normal mice,while 15 mg·kg-1 ketamine group could significantly increase the number of standing of mice?P<0.05?.Western blot showed that the expression of p-ERK,BDNF,p-mTOR and synapsin protein decreased in the corticosterone group.Compared with corticosterone group,the expression of p-ERK,BDNF,p-mTOR and synapsin protein in combination group and 0.1?M ketamine group increased,but there was no significant difference.Compared with the solvent control group,there was no significant difference in the expression of p-ERK and synapsin in the cerebral cortex between the combination group and the ketamine 15 mg·kg-1 group.Conclusion:The combination of ketamine and butylphthalide can protect PC12 cells damaged by corticosterone and enhance the synaptic plasticity of neurons.In animal experiments,it is found that the combination of ketamine and butylphthalide can not significantly reduce the accumulated immobility time in the behavior of mice,while the combination of ketamine and butylphthalide can reduce the forced swimming and tail suspension of mice.In the open field experiment,it was found that ketamine 15 mg·kg-1could increase the number of times of standing,but the combined group did not.It is suggested that ketamine combined with butylphthalide can produce antidepressant effect,and its mechanism is related to the activation of BDNF-mTOR signal pathway. |
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