| Objective:To study the effect of Bupleurum plus Dragon Bone and Oyster Decoction on the expression of 5-HT2 AR and CaMKⅡα in hippocampus of depression rats,and to explore the antidepressant mechanism of Bupleurum plus Dragon Bone and Oyster Decoction.Method:Fifty SPF male SD rats were randomly divided into two groups: groupⅠwas a blank group,10 were given normal feeding,and group II was a model.CUMS combined with orphanage to establish a depression model,and forced swimming,tail suspension,and open field experiments to evaluate depression in rats.Randomly divided into 3 groups(model group,Bupleurum plus Dragon Bone and Oyster Decoction group,fluoxetine group)to determine the body weight and behavioral changes of rats;HE staining to observe the changes in rat hippocampus;-MRNA expression of HT2 AR and CaMKⅡα;immunohistochemical method to detect the expression of 5-HT2 AR and CaMKⅡα protein.Result:1.Effect on weight:After the model building is completed,the weight of the blank group is significantly higher than that of the model group,and the difference is significant and statistically significant(P <0.05).After 4 or 6 weeks of gavage,the weight of each group increased,and there was no difference between the groups(P> 0.05).After 8 weeks of gavage,there was no significant difference between the Bupleurum plus Dragon Bone and Oyster Decoction group and the fluoxetine group and the model group(P> 0.05).Compared with the fluoxetine group,the Bupleurum plus Dragon Bone and Oyster Decoction group had no significant weight gain and no statistical significance(P> 0.05).2.Behavioral testing:The effect of the drug on the tail suspension immobility time of depression rats:Before administration,the model group prolonged the tail suspension immobility time compared with the blank group(P <0.05).After 4 weeks of administration,there was no significant change in the immobility time of the tail of Bupleurum plus Dragon Bone and Oyster Decoction group and model group,and the difference was not statistically significant (P> 0.05).Compared with the model group,the fluoxetine group had shorter tail-hanging immobility time,and the difference was statistically significant(P <0.05).Compared with the fluoxetine group,the Bupleurum plus Dragon Bone and Oyster Decoction group had no statistically significant difference(P> 0.05).After 8 weeks of administration,the Chaihu Jia longgu muli decoction group and fluoxetine group had shorter tail dwelling time than the model group,and the difference was statistically significant(P <0.05).Compared with the fluoxetine group,the Bupleurum plus Dragon Bone and Oyster Decoction group had no significant difference(P> 0.05).The effect of drugs on swimming float time of depressed rats:The swimming float time of the model group before administration was longer than that of the blank group,the difference was statistically significant(P <0.05).Four weeks after the administration,the Bupleurum plus Dragon Bone and Oyster Decoction group and the fluoxetine group had shorter swimming float times than the model group,and the difference was statistically significant(P <0.05).Compared with the fluoxetine group,the Bupleurum plus Dragon Bone and Oyster Decoction group was significantly different(P <0.05).After 8weeks of administration,the Bupleurum plus Dragon Bone and Oyster Decoction group and the fluoxetine group had shorter swimming and floating times than the model group,and the difference was statistically significant(P <0.05).Compared with the fluoxetine group,the Bupleurum plus Dragon Bone and Oyster Decoction group had no significant difference(P>0.05).The effect of drugs on the open field experiment in depressed rats:The total distance of the model group before administration was reduced,the rest time was increased,and the wall-standing time was reduced.The difference was statistically significant(P <0.05).After 4 weeks of administration,the total distance of the fluoxetine group was significantly increased compared with the model group(P <0.05).Compared with the model group,the rest time of Bupleurum plus Dragon Bone and Oyster Decoction group and fluoxetine group was significantly shortened,which was statistically significant(P<0.05).The fluoxetine group had significantly shorter rest time than Bupleurum plus Dragon Bone and Oyster Decoction group,with statistical significance(P <0.05).There was no significant difference between the Bupleurum plus Dragon Bone and Oyster Decoction group and the fluoxetine group compared with the model group during the climbing time and standing time(P> 0.05).After 8 weeks of administration,the total distance of the Bupleurum plus Dragon Bone and Oyster Decoction group and the fluoxetine group was significantly increased compared with the model group,with statistical significance(P<0.05).Compared with the Bupleurum plus Dragon Bone and Oyster Decoction group,the total distance of fluoxetine group increased significantly with a significant difference(P<0.05).Compared with the model group,the rest time of Bupleurum plus Dragon Bone and Oyster Decoction group and fluoxetine group was significantly shortened,and the difference was significant,with statistical significance(P <0.05).Compared with the Bupleurum plus Dragon Bone and Oyster Decoction group,the rest period was significantly shorter in the fluoxetine group(P <0.05).Compared with the model group,the Bupleurum plus Dragon Bone and Oyster Decoction group and the fluoxetine group had no significant difference in standing time(P> 0.05).There was no significant difference in wall-climbing time between the fluoxetine group and the Bupleurum plus Dragon Bone and Oyster Decoction group(P>0.05).3.Hippocampus HE staining:The pyramidal cells of hippocampus in model group were scattered,irregular in shape and pyknotic in nucleus.After the intervention of Bupleurum plus Dragon Bone and Oyster Decoction and fluoxetine,the pyramidal cells of the hippocampus of the rats were arranged in order,occasionally with hyperchromatic nuclei and large cell bodies.4.Real-time PCR to detect the expression of 5-HT2 ARm RNA and CaMKⅡαm RNA:After 8 weeks of drug intervention,the expression of 5-HT2 AR and CaMKⅡα m RNA in Bupleurum plus Dragon Bone and Oyster Decoction group and fluoxetine group increased.Compared with the model group,the difference was significant P <0.05.5.Immunohistochemical method was used to detect the expression of 5-HT2 AR and CaMKⅡα:After 8 weeks of drug intervention,the expression of 5-HT2 AR and CaMKⅡαm RNA in Bupleurum plus Dragon Bone and Oyster Decoction group and fluoxetine group increased.Compared with the model group,the difference was significant <0.05.Conclusion:1.Depression modeling may slow the weight gain of rats,and antidepressant treatment does not change the growth trend of rats.2.Bupleurum plus Dragon Bone and Oyster Decoction can increase the time of autonomous activity in rats,fluoxetine is more effective than Bupleurum plus Dragon Bone and Oyster Decoction.3.The anti-depression of Bupleurum plus Dragon Bone and Oyster Decoction and fluoxetine canimprove the structure and distribution of pyramidal cells in the hippocampus,there by improving the plasticity of hippocampal neurons.4.The anti-depression mechanism of Bupleurum plus Dragon Bone and Oyster Decoction group can increase the 5-HT2 AR and CaMKⅡα m RNA levels in the hippocampus of depressed rats and increase the expression of 5-HT2 AR and CaMKⅡα proteins. |
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