| BackgroundDepression is a disease with clinical manifestations of continuing depression, lack of sense of pleasure, loss of interest, thinking and cognitive retardation and affective disorders and other symptoms. Sleep deprivation is an effective method of rapid antidepressant effect, but the mechanism is unclear. Combination of antidepressants and sleep deprivation are often able to obtain a better antidepressant effect more consolidation in clinical practice, and are easily accepted by patients. Clomipramine is tricyclic antidepressants, its efficacy in the treatment of various types of depression and anxiety disorders, its5-serotonin reuptake strong blocking effect. In this study, we establish chronic unpredictable mild stimulation of rat model of depression and observe effect of sleep deprivation, clomipramine on CREB, PKA expression, neuronal apoptosis and behavior of rat depression model.Objective(1) To establish chronic unpredictable mild stress rat model of depression and evaluate their behavior. To observe the behavioral improvement after sleep deprivation, clomipramine, and their combination in model of depression of rats.(2) By detecting hippocampal CREB, PKA expression and neuronal apoptosis related genes Bcl-2and Bax for study mechanism of sleep deprivation, clomipramine, and their combined antidepressant.MethodsSelect70male SD rats with homogeneity, weighing310-420g, of which60rats were used to construct depression model, and the other10served as controls. Depression model established using28days of chronic unpredictable mild stress stimulation. Weight model rats’weighing, sucrose consumption test, open-field test and water maze test were used to detect behavior of depression rats model. Select40depression rats model were randomly divided into normal group except for the other groups as follows:normal group (10, A group), depression group (10, group B), clomipramine group (10, group C), sleep deprivation group (10, D), sleep deprivation combined clomipramine group (10, E group). Sleep deprivation time:36h continuous sleep deprivation was performed at1,8,15,22th days. Clomipramine administered at a dose of5mg/kg,1/d, continued to28d. Detect the behavior of rats in each group changes the situation in28days after the adoption of the above methods. Real-time quantitative PCR, Western blot was used to detect the expression, of hippocampal GA3region of CREB, PKA, Bcl-2and Bax mRNA and protein. Detect the neuronal apoptosis using the TUNEL method.Results(1) After28days of chronic unpredictable mild stimulation, the model group average weight was (341.1±23.9) g, significantly lower than the control group (448.5±39.2) g (P<0.05). Average water consumption of sucrose in model group was (6.4±0.4) ml/100g, significantly lower than the control group (13.6±0.8) ml/100g (P<0.05). Open-field test results show that the latency, the number of horizontal movement distance and erected times in model rats were (3.9±0.9) s,(205,7±18.3) cm and (5.2±1.3) times, significantly lower than the control group.(2) After28days of different interventions in each group, body weight of A and E group were significantly increased, group B was no significant increased, the rate of weight increase in group C and D were less than the group E. Sucrose water test and open field test and water maze test in group E showed a significant improvement, and group C and D group improved less than group E, but the difference between group C and D groups had not statistically significant (P>0.05).(3) CREB expression in group A and E were significantly higher than the other three groups (P<0.05), the difference between the two groups was not statistically significant (P>0.05). Expression of PKA was lowest in Group B, which was significantly lower than the other four groups (P<0.05). Bcl-2gene and protein expression level of group B and D was significantly lower than group A and E (P<0.05), while Bcl-2expression between Groups C and D groups showed no significant difference (P>0.05). Bax gene and protein expression in group B was significantly higher than other groups, pairwise comparison found that C, D, E groups were not statistically significant in the Bax gene and protein expression compared with group A (P>0.05). Sleep deprivation apoptotic rate was the highest (55±6)%, significantly higher than the control group (14±3)%and the combination group(19±4)%.Conclusions(1) Sleep deprivation combined clomipramine can improve behavior of depression model of rats.(2) Sleep deprivation combined clomipramine can effectively increase CREB, PKA and Bcl-2expression and reduce Bax expresson in CA3region of the depression model rats hippocampus.(3) Sleep deprivation combined clomipramine can reduce neuronal apoptosis in rat hippocampal CA3region of depression model. |
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