| Depression has the characteristics of high prevalence rate,high recurrence rate and high suicide rate,so it has become one of the frequently-occurring diseases that seriously affect human life and health.At present,western medicine is the main clinical treatment,but it has serious side effects,poor tolerance and high price.Traditional Chinese medicine in the treatment of depression has the advantages of multi-target and integrity,small adverse effects and high safety,which has attracted the attention of more and more researchers.Traditional Chinese medicine Bupleurum has the effect of soothing the liver and relieving depression.It has a history of hundreds of years in the treatment of depression,and it is frequently used in antidepressant prescriptions.The traditional Chinese medicine Bupleurum is divided into North Bupleurum and Southern Bupleurum.Studies have shown that compared with North Bupleurum,South Bupleurum is better at soothing the liver and relieving depression,but the antidepressant mechanism has not been explained.In addition,vinegar can enhance the effect of soothing the liver and relieving depression,but its specific mechanism remains to be revealed.Therefore,this paper explores the antidepressant effect and mechanism of raw and vinegar products of Bupleurum from the perspective of metabonomics and intestinal flora diversity,so as to provide scientific basis for clinical application of Bupleurum in the treatment of depression.Firstly,the depression-related model of rats was established by chronic unpredictable mild stimulation combined with isolation.The efficacy of Nanchaihu and its vinegar products were evaluated by general behavioral observation,behavioral tests(sugar water preference rate and open field test),and the contents of dopamine(DA),serotonin(5-HT)and norepinephrine(NE)in the brain.The results showed that the number of open field activities,the preference rate of sugar water and the content of monoamine neurotransmitters in the brain of the rats after 28 days of modeling were significantly lower than those of the blank control group(P<0.05).However,after 21 days of treatment with raw Bupleurum and vinegar products,the abovementioned indexes increased significantly(P<0.05).All the indexes of rats in the South Bupleurum vinegar group were better than those in the raw group,but the difference was not statistically significant.Furthermore,the profile of serum metabolites of rats in each group was measured by UPLC-Q-TOF/MS technique,compounds were identified by ProgenesisQI software,and differential metabolites and metabolic pathways related to depression were screened by Metaboanalyst online website to study the changes of metabolites in rats before and after treatment with raw Bupleurum and vinegar products.Results 27 differential metabolites associated with depression were obtained.Include:cholic acid,chenodeoxycholic acid,sphingosine-1-phosphate,taurocholic acid,LysoPC(18:0),(12,13-EpOME),11βhydroxyprogesterone,testosterone,tryptamine,sphingomyelin,arachidonic acid,phosphatidylethanolamine,cholesterol sulfate,prostaglandin E2,LysoPC(18:1),5-HETE,phospholipid choline,L-tryptophan,L-phenylalanine,testosterone glucuronic acid,19hydroxytestosterone,testosterone,Phosphatidyl serine,dehydroepiandrosterone sulfate,glycine chenodeoxycholic acid,bezoar deoxycholic acid,glycolic acid and 5-hydroxy-Ltryptophan,etc.The results showed that arachidonic acid metabolism,α-linolenic acid metabolism,sphingolipid metabolism,tryptophan metabolism,phenylalanine tyrosine tryptophan biosynthesis and glycerol phospholipid metabolism were disturbed in the model rats.However,18 differential metabolites could be significantly recalled in the treatment of South Bupleurum,and 21 differential metabolites could be significantly recalled in the treatment of South Bupleurum vinegar,which involved two kinds of metabolic pathways,such as amino acid metabolism and lipid metabolism.On this basis,the single molecule real-time sequencing method based on PacBio sequencing platform was used to sequence the fecal microflora of depression model rats before and after treatment of raw and vinegar products of Bupleurum chinense.Then the sequence was filtered and clustered,and Alpha diversity analysis,Beta diversity analysis and species abundance analysis were carried out.The results showed that compared with the normal rats,the structural diversity and abundance of intestinal flora in the model group decreased,while the raw Bupleurum and vinegar products could play a callback role.At the genus level,compared with normal rats,the abundance of Muribaculum in the model group increased significantly,while the abundance of Prevotella,Lactobacillus,Monoglobus,Christensenelacea-R-7-group,Lachnospiraceae_NK4A136_group,Escherichia coli/Shigella and Ruminococcaceae_UCG-013 decreased significantly(P<0.05).Compared with the model group,the abundance of Prevotella,Christensenelacea-R-7-group,Monoglobus,Lactobacillus,Muribaculum and Ruminococcaceae_UCG-013 could be significantly adjusted in the raw and vinegar products of Bupleurum.In addition,the abundance of Rosella could be significantly adjusted in the raw products of Bupleurum,and the abundance of Lachnospiraceae_NK4A136_group could be significantly adjusted in the vinegar products of Bupleurum chinense.In addition,it was also found that at the seed level,compared with normal rats,the abundance of Bacteroidales_bacterium,Clostridium_sp.and Rombousia_idedls in model rats increased significantly(P<0.05),Monoglobus-pecitinilyticus,while the abundance of Escherichia coli decreased significantly(P<0.05).However,both raw and vinegar products of Bupleurum chinense could significantly adjust the abundance of Monoglobus_pecitinilticus and Rombousia-idedls(P<0.05),indicating that both raw products and vinegar products of Bupleurum chinense could effectively improve the functional disorder of intestinal flora in rats caused by depression.Finally,the Pearson correlation coefficient method was used to analyze the correlation between the peak area of metabolic biomarkers and the abundance of intestinal microflora.The results showed that among the top 30 abundant bacteria,Lactobacillus and Clostridium_sensu_strictol were involved in tryptophan metabolism,Klebsiella,Bacteroides and Prevotella were involved in sphingolipid metabolism,Rosella,Lachnospiraceae_NK4A136_group and Ruminococcaceae_UCG-013 were involved in bile acid synthesis,Lachnospiraceae_NK4A136_group was involved in glycerolipid metabolism,Muribaculum was involved in steroid synthesis.The imbalance of intestinal microflora may cause the disorder of host amino acid metabolism and lipid metabolism pathway.To sum up,raw and vinegar products of Bupleurum chinense may have antidepressant effects by regulating the abundance of Lactobacillus,Clostridium_sensu_strictol,Rosella,Lachnospiraceae_NK4A136_group,Klebsiella,Bacteroides,Prevotella,Ruminococcaceae UCG-013 and Muribaculum,and then affecting amino acid metabolism and lipid metabolism,and finally affecting the synthesis of neurobiofilms and monoamine neurotransmitters in the brain. |
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