| Objective: This study adopts the chronic unpredictable mild stress depression model rats as the research object, from the perspective of behavioral scienceto discuss the effect of antidepressant and mood stabilizing by acupuncture method, from the change of rat hippocampal GLU and GABA content and NMDA-receptor subunits protein expression to explore the part of the central mechanism of acupuncture treatment of depression, so as to provide a scientific basis and foundation for the generalization and application of acupuncture treatment of depression, and provide new ideas for the pathogenesis research of depression.Methods: Choose 120 SPF adult SD rats, were randomly divided into blank control(BC) group, the depression model(DM) group, valproate treatment(VT) group,fluoxetine treatment(FT) group, the acupuncture treatment(AT) group and the non-acupoint acupuncture treatment(NAAT) group, each group of 20, male and female half and half. In addition to the blank control group,the other five groups of animals were used in the chronic unpredictable mild stress(the chronic unpredictable mild stress, CUMS)method to make depression model. Meanwhile, the NAAT acupunctured non-acupoint of rats; the AT acupunctured the "Baihui", "Neiguan" and "Sanyinjiao" points; the VT lavaged the valproate fluid and the FT lavaged fluoxetine fluid.Once a day for 21 days. All the animals were weighed at the day 1 and day21,and determinated sugar water consumption, to compared the changes of weights and consumption of sucrose water of each group. Observed the changes of behavior of rats at different hours:before the experiment, the day 7, the day 14, the day 21.Take each rat hippocampus after the experiment, using high performance liquid chromatography(HPLC) to test the content of each sample of Glu, GABA and detect protein expression levels of rat hippocampal NR1 and NR2A(NMDA-receptor subunits) by Western Blot method.The results:1. The influence of weight Before building model, there were no significant differences between groups of rats(P > 0.05). The day 21, the weight of DM was significantly lower than the BC(P< 0.01). In terms of weight mong the VT,the FT and AT,compared with the BC, there were no significant differences(P > 0.05). But VT’s weight increased significantly(P< 0.05), rather than the AT’s.Between the NAAT and the DM,it was no significant difference(P > 0.05).2. The influence of behavioral science2.1 The consumption of sugar water Before the experiment, the intake of 1% sucrose water each rat is on the basic level, and there were no statistically significant differences(P > 0. 05). At the day 21,the intake of 1% sucrose water of the DM was significantly lower than the BC’s,and the difference was statistically significant(P < 0.01). Three differences were statistically significant among the FT, the VT and the AT, compared with the DM(P< 0.01), and the comparison between the 3 groups had no statistical differences(P >0.05).2.2 The Open field test After the experiment,we tested the explorative activity each rat by the Open-field method and recorded activities within 3 minutes. Results show that, compared with the BC, the DM’s scores of horizontal and vertical movement were significantly lower(P < 0.01).Compared with the DM, the scores of horizontal and vertical movement among the FT, the VT and AT, were significantly increased, and the differences were statistically significant(P < 0.01). Compared with the AT, the scores of horizontal movement between the FT and the VT were significantly lower, and the difference was statistically significant(P < 0.01).Compared with the FT,the scores of vertical movement between the VT and the AT were significantly lower, and the difference was statistically significant(P < 0.01).Compared the VT with the AT,there was no significant difference(P > 0.05).3. The influence of the testing results of immunohistochemical staining3.1 The expression of GLU and GABA positive neurons in hippocampal CA1 area Compared with the BC, the expression of GLU positive neurons of the DM in hippocampal CA1 area was increased significantly(P < 0.01), and the expression of GABA positive neurons was decreased significantly(P < 0.01), suggesting that:making depression mold can cause CUMS’s rats hippocampal CA1 area GLU content increase and GABA content decrease.Compared with the DM, the expressions of GLU positive neurons both the VT and the AT were decreased significantly(P <0.01), and the expressions of GABA positive neurons both the VT and the AT were no significant change(P > 0.05),and there was no significant difference between the VT and the AT, suggesting that: the VT and the AT can effectively adjust the growth of GLU content in hippocampal CA1 area caused by CUMS, while the FT and the NAAT had no effect.3.2 The expression of GLU and GABA positive neurons in hippocampal CA3 area Compared with the BC, the expression of GLU positive neurons of the DM in hippocampal CA3 area was increased significantly(P <0.01), and the expression of GABA positive neurons was decreased significantly(P <0.01), suggesting that:making depression mold can cause CUMS’s rats hippocampal CA3 area GLU content increase and GABA content decrease.Compared with the DM, the expressions of GLU positive neurons both the VT and the AT were decreased significantly(P <0.01),but the expression of the VT declined significantly(P <0.05), rather than the AT’s, and the difference was statistically significant(P <0.05). Compared with the DM, the expressions of GABA positive neurons both the VT and the AT were significantly increased(P <0.01, P <0.05), but it was no significant difference between the VT and the AT(P >0.05).suggesting that: the VT and the AT can effectively adjust the growth of GLU content and the decrease of GABA content in hippocampal CA3 area caused by CUMS, while the FT and the NAAT had no effect.In terms of cutting down the content of GLU,the VT is superior to the AT.4. The influence of the testing results of subunits of NMDA receptor by Western Blot method Compared with the BC, the expressions of NMDA-receptor subunit NR1 and NR2 A protein of the DM were increased significantly(P < 0.01).Compared with the DM, the expressions of NMDA-receptor subunit NR1 and NR2 A both the VT and the AT were decreased significantly(P <0.01, P <0.05),then there was no significant difference between the VT and the AT(P>0.05).And there were no significant differences among the FT,the NAAT and the DM(P>0.05).There were no significant differences on GABAAR and NR2 B protein expression levels among 6 groups(P >0.05).In conclusion:1. Acupuncture points has good treatment effect on depression model rat,and it is the same with fluoxetine and valproic acid sodium. While acupuncture non-acupoints had no effect.2. It may be associated the antidepressant effect of the acupuncture points with adjusting the imbalances of GLU and GABA content in hippocampus brain regions.But antidepressant mechanism of fluoxetine has nothing to do with GLU/GABA system.3. It may be associated the antidepressant effect of the acupuncture points with reducing hippocampal NMDA-receptor subunits NR1 and NR2 A protein expression, so as to lower the toxicity of excitatory GLU. |
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