Study On The Mechanism Of The Effects Of Oviductus Ranae On Depressed Model Mice

Objective: To replicate the mouse depression model by chronic unpredictable mild stress,and to detect changes in related behavioral indicators,apply enzyme-linked immunosorbent assay,flow cytometry,HE staining,and Western blotting to determine brain and serum biochemical indicators and hippocampal tissue,the morphological changes and the expression levels of related proteins,evaluate the antidepressant effect of Oviductus Ranae and clarify its mechanism.Methods: 1.60 healthy male ICR mice,equally divided into blank control group,model control group,fluoxetine positive group,and Oviductus Ranae high,medium and low dose group.Except the blank control group,all the mice in the other groups received chronic mild and unpredictable stress for 28 days.Dexamethasone 3mg/kg,Oviductus Ranae high800 mg/kg,Oviductus Ranae medium 400 mg/kg and Oviductus Ranae low 200mg/kg.2.Detection indexes :(1)After the last dose,the mice in each group were subjected to behavioral test;(2)The contents of 5-ht,NE,DA,CRH,ACTH and CORT in the serum were measured by ELISA.(3)Hippocampal tissues of mice in each group were cut and ground to make cell suspension,which was filtered through 300 mesh nylon mesh,and the apoptosis of hippocampal cells was detected by flow cytometry.(4)Hippocampal tissue was taken,investigate the effect of oviductus ranae on hippocampal structure in depressed mice by HE.(5)The remaining hippocampal tissues were preserved at-80?,and the expression of GR,MR,BDNF,Trk B and CREB proteins in the hippocampal tissues were detected by western blot.Results: 1.Behavioral test results: Compared with the blank control group,weight loss occurred in the model control group;2.Sugar water consumption decreased;Tail suspension experiment and forced swimming experiment were prolonged;In the open field experiment,the movement time,movement distance and center stopping time decreased,while the corner stopping time increased(P<0.05).Compared with the model control group,the weight of fluoxetine positive group and Oviductus Ranae group increased significantly.Increased consumption of sugar water;Tail suspension experiment and forced swimming experiment shorten the resting time;The movement time,movement distance and center stopping time of the open-field experiment increased,while the corner stopping time decreased(P<0.05).2.ELISA detected the content of serum related indicators: Compared with the blank control group,the content of CRH,ACTH and CORT in the serum of the model control group increased significantly(P<0.01).The contents of 5-ht,NE and DA were significantly reduced(P<0.01).Compared with the model control group,the content of CRH,ACTH and CORT in fluoxetine positive group and Oviductus Ranae high,medium and low dose group decreased significantly(P<0.05).The contents of 5-ht,NE and DA were significantly increased(P<0.05).3.Detection of hippocampal cell apoptosis: Compared with the blank control group,the proportion of hippocampal cell apoptosis in the model control group increased significantly(P<0.01).Compared with the model control group,the apoptosis rate of hippocampal cells in fluoxetine positive group and Oviductus Ranae group was significantly decreased(P<0.05).4.Hippocampal pathological changes: The hippocampus cells of the blank control group were densely and neatly arranged.Compared with the blank control group,the hippocampus cells in the model control group were obviously atrophic,disordered and reduced in volume.Compared with the model control group,fluoxetine positive group and Oviductus Oanae dose group were improved to different degrees.5.Protein expression in western blot detection: Compared with the blank control group,the expressions of GR,MR,BDNF,Trk B and CREB in the model control group were significantly decreased(P<0.01).Compared with the model control group,the expression levels of GR,MR,BDNF,Trk B and CREB in fluoxetine positive group and Oviductus Ranae dose group were significantly increased(P<0.01).Conclusion: 1.The model of chronic unpredictable mild stress simulates the depression caused by long-term acceptance of chronic stress.Similar to the pathogenesis of human depression,Oviductus Ranae can effectively improve the behavioral changes caused by chronic unpredictable mild stress.2.Oviductus Ranae could effectively increase the contents of monoamine neurotransmitter 5-ht,NE and DA in the serum of mice with depression.The contents of CRH,ACTH and CORT were decreased.3.Oviductus Ranae can inhibit the apoptosis of hippocampal cells,reduce the damage of hippocampal tissues,and improve the pathological changes of hippocampal tissues;4.Oviductus Ranae can increase the expression of GR and MR proteins,inhibit the hyperactive HPA axis through negative feedback regulation,and improve the expression of BDNF,Trk B and CREB proteins,and may play the neuronal protection role of brain-derived neurotrophic factor through this pathway.