| BackgroundDepression is a chronic recurrent disease,with persistent low mood and cognitive dysfunction as the main clinical features,including attention and anhedonia,lonely depression,sleep disorders,movement and mental retardation,and even suicidal thoughts.The disease has an extremely high incidence rate,recurrence rate,self-harm rate and suicide rate,and has become a prominent problem affecting the global public health safety.In recent years,with the incidence of depression and more rapid,people’s physical and mental health has been profoundly affected,and the prevention and treatment of depression is worth paying attention to.The Notch signaling pathway,which is closely related to the pathogenesis of depression,plays an important role in regulating the proliferation,regeneration and differentiation of nerve cells.Liver and spleen deficiency is a common TCM type of depression,and studies have shown that chronic binding stress can successfully replicate the depression model of liver and spleen deficiency in depression.The "square certificate correspondence"relationship between xiaoyaosan and liver depression and spleen deficiency syndrome has been repeatedly proved by a large number of literature,experiments and clinical research institutes.Xiaoyaosan focuses on the overall syndrome treatment of depression,liver depression and spleen deficiency,and monomer saikosaponin A further focuses on the regulatory effect of the active components.Therefore,based on Notch signaling pathway,the neuroregulation and intrinsic action mechanism of xiaoyaosan and saikosaponin A on depression can be investigated.ObjectiveThis study aims to explore the effect of xiaoyaosan and saikosaponinA in rats from Notch pathway,and the number of positive cells,and the expression of key molecular signal on the Notch pathway,which provides scientific experimental basis for the prevention and treatment of depression in traditional Chinese medicine.MethodSixty healthy male SD rats were selected,weighing from 220 to 250 g.After 7 days of adaptive rearing,rats were divided into 5 groups by body weight,including normal group,model group,saikosaponinA group,fluoxetine group and xiaoyaosan group.Each group consisted of 12 individuals.Chronic bound stress was performed for 21 days in rats except for the normal group.The gavage intervention was performed on saikosaponinA group,fluoxetine group and xiaoyaosan group.After the molding was completed,the behavioral experiment(sugar-water preference experiment and open field experiment)and the materials were used.The depressed rat model was evaluated by analyzing rat changes in general macroscopic signs,body weight,food intake and behavior.The morphological changes of hippocampus were observed by HE staining,the number of positive cells was observed by immunofluorescence,and the protein and gene expression of Notch1,Jagged1 and Hes1 were detected in the Notch pathway of rat hippocampus by protein immunoblotting(western blot)and real-time PCR.Result(1)A rat model of liver depression and spleen deficiency,mental depression,reduced activity,motor delay,hair dryness and no hair appeared.The intake,weight and gain were significantly lower than the normal group(P<0.05).In the model group,the preference rate,standing times,modification times,entering the central area and the total distance of exercise were lower than those of the normal group(P<0.01).SaikosaponinA,fluoxetine and xiaoyaosan all improved on depressive behavior.(2)In CA1 of CA3 and DG,Compared with the normal group,the number of BrdU/NeuN and BrdU/DCX positive cells was significantly decreased in the DG area of the hippocampus in the model group(P<0.01).Compared with the model group,the number of BrdU/NeuN positive cells in the saikosaponinA group,fluoxetine group and xiaoyaosan group increased significantly(P<0.01).Although the number of BrdU/NeuN positive cells was increased in each administered group,it was still different compared with the normal group(P<0.05).The number of BrdU/DCX positive cells increased in the saikosaponinA group(P<0.05);The number of BrdU/DCX positive cells increased in the xiaoyaosan group compared significantly(P<0.01);Although the number of positive cells could be increased in fluoxetine,the difference was not significant compared with the model group(P>0.05).It was still different from the normal group(P<0.05).(3)RT-PCR results showed that the gene expression of Notchl,Jaggedl,and Hes1 decreased significantly compared with the normal group(P<0.01).Compared with the model group,cccccc group,fluoxetine group and xiaoyaosan group can upregulate Notchl and Hesl(P<0.01);Fluoxetine group and xiaoyaosan group can upregulate Jaggedl(P<0.05).(4)The western blot results showed that Notch1,Jagged1,and Hes1 protein expressions were significantly decreased in the model rats compared with the normal group(P<0.01).Significant difference in Notchl protein expression in the xiaoyaosan group of rats compared to the model group(P<0.01).Jaggedl protein expression was significantly increased in saikosaponinA,fluoxetine and xiaoyao group(P<0.01).The protein expression of Hes1 was significantly increased in the saikosaponinA and xiaoyaosan group(P<0.01).In the Notch1 and Jaggedl protein expression,each administration group still performed significantly differently compared with the normal group(P<0.05orP<0.01).In Hes1 protein expression,the difference insaikosaponinA and fluoxetine compared with the normal group(P<0.01).Conclusion(1)21-day chronic binding stress can successfully replicate the rat model of liver depression and spleen deficiency depression.(2)Chronic stress shows nerve damage in the hippocampus and the decline of nerve cell proliferation and differentiation ability in the depressed rat model.After drug intervention and treatment,the nerve damage in the hippocampus can be repaired to some extent.The treatment effect of xiaoyaosan is better than saikosaponinA.This suggests that xiaoyaosan and saikosaponinA may exert antidepressant effects by regulating neural cell function in the hippocampus.(3)Based on the Notch pathway,xiaoyaosan and saikosaponinA can significantly upregulate the gene and protein expression of key signaling molecules in the pathway,suggesting that xiaoyaosan and saikosaponinA may play a hippocampal neuroprotective role through the Notch pathway to improve the depressive symptoms of liver depression and spleen deficiency.The effect of saikosaponinA is weaker than that of xiaoyaosan compound. |
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