| Anxiety is a common chronic mental disorder,characterized by depression,lack of interest,guilt,often accompanied by excessive anxiety,restlessness,insomnia,muscle tension and other physiological awakening symptoms.Anxiety disorder is characterized by high morbidity,high mortality and high recurrence rate.It seriously affects people’s production and quality of life,and also brings huge economic burden to individual families and society.The pathogenesis of anxiety disorder is very complex.In the past,there have been many researches on its pathogenesis in the aspects of genetics,immunity and endocrine.Genome-wide studies have confirmed the genetic characteristics of anxiety disorder,but this situation may also be triggered by environmental factors.Although antidepressants have a good effect in the treatment of anxiety,such as selective serotonin reuptake inhibitors(SSRIs),but patients often have symptoms of relapse or adverse reactions and other shortcomings.Schizandra chinensis(Turcz)Baill,as a traditional tranquilizing drug in China,its main pharmacological activity is lignan,which has a variety of pharmacological effects,such as scavenging free radicals in vivo,protecting liver,improving immunity,central inhibition and tranquilizing effects.In clinic,the processed products are mostly used as medicine.After processing,the efficacy of Schisandra chinensis has changed a lot.The main reason is that the effective ingredients of Schisandra chinensis have changed after processing with yellow wine.At present,the research on raw Schisandra chinensis is mostly carried out at home and abroad,but the research on the anti-anxiety mechanism of wine processed Schisandra chinensis is rarely reported.In addition,the latest research found that there is a close relationship between intestinal flora and the production of anxiety disorder.Intestinal flora can affect the metabolic phenotype of human body,which is of great significance for maintaining human health.It may be a new target for the treatment of anxiety,therefore,based on metabonomics and intestinal flora,the anti anxiety mechanism of Schisandra chinensis was explored.The specific research contents are as follows:1.Based on UPLC-Q/TOF-MS,12 lignans in the water extract of Schisandra chinensis before and after processing were analyzed quantitatively.The results showed that the contents of 6-o-phthaloyl-gomisin o,schisandrin,gomisin D,schisandrin A,gomisin T,schisandrin B and schisandrin C increased,while the contents of schisandrin A,schisandrin B,schisandrin C,schisandrin D and new schisandrin B decreased.2.In this study,the internationally recognized chronic unpredictable stress(CUS)method was used to induce anxiety rat model,and the field experiment(OFT),elevated cross maze experiment(EPM),suspended tail experiment(TST)and forced swimming experiment(FST)were used as evaluation indexes.After 5 weeks of stimulation,the behavior of rats in the model group showed that the behavior of independent exploration in the field environment was reduced and increase of residence time,tail suspension time and swimming time in the closed arm of cross maze indicated that the anxiety model induced by CUS was established successfully.3.Based on UPLC/LTQ-orbitrap-MS technique,the non-target metabonomics of plasma and brain samples of anxiety rats and control rats was studied.The results showed that OPLS-DA analysis showed that the samples of plasma and brain tissue of anxiety rats had a significant trend of classification.49 and 23 different metabolites of plasma and brain tissue related to anxiety were screened by univariate statistical analysis.Through further analysis of these differential metabolites,we found that anxiety rats were mainly accompanied by metabolic abnormalities such as glycerol phospholipid metabolism,cholic acid metabolism and sphingolipid metabolism.4.16S rRNA high-throughput sequencing was used to analyze the intestinal microflora structure of anxiety rats and control rats.The diversity and richness of intestinal microflora in anxiety rats were significantly different from those in the control group.Beta diversity analysis showed that there was a significant difference in the intestinal microflora structure between anxiety rats and control rats.Further studies showed that the main feature of this imbalance was that some beneficial bacteria,such as Lactobacillus,had significantly increased the intestinal abundance in anxiety rats.These changes may be related to the occurrence and development of anxiety.5.The anxiety model induced by CUS was used to evaluate the antianxiety effect of water extracts from raw Schisandra chinensis and wine processed Schisandra chinensis.The results show that,through behavioral experiments,raw and wine processed Schisandra chinensis can significantly increase the independent exploration behavior in the field experiment of anxiety rats,increase the stay time of anxiety rats on the open arm of the elevated cross maze,and significantly reduce the immobility time of anxiety rats in tail suspension and forced swimming,which suggests that raw and wine processed Schisandra chinensis can improve the anxiety behavior of rats,wine processed Schisandra chinensis is better than raw Schisandra chinensis.The results of ELISA,RT-q PCR,Western blot and immunofluorescence showed that wine processed Schisandra chinensis can significantly reduce the content of lactate in plasma,the expression of GPR81 mRNA in liver and the expression of TNF-α in hippocampus,wine processed Schisandra chinensis can improve the expression of S1PR2 in hippocampus and up regulate the expression of AC and PKA protein in liver.This suggests that wine processed Schisandra chinensis may have anti anxiety effect by regulating lipid metabolism and reducing inflammatory factors in hippocampus.Secondly,based on the technology of UPLC/LTQ-orbitrap-MS,the above-mentioned plasma samples of rats fed with Schisandra chinensis were analyzed by metabonomics,to further explore the effect of Schisandra chinensis on the overall metabolic spectrum of anxiety rats and its related mechanism.The results showed that there were significant differences in plasma metabolism between the anxiety group and the raw Schisandra chinensis group,and between the anxiety group and the wine processed Schisandra chinensis group.Among them,compared with the anxiety group rats,the raw Schisandra chinensis regulated 29 endogenous metabolites,the wine processed Schisandra chinensis regulated 36 endogenous metabolites,the raw Schisandra chinensis and wine processed Schisandra chinensis regulated 25 endogenous metabolites at the same time.In conclusion,12 lignans in Schisandra chinensis before and after processing were quantitatively analyzed based on UPLC-Q/TOF-MS technology,and then the endogenous differential metabolites related to anxiety plasma and brain tissue were screened and analyzed by UPLC/LTQ-orbitrap-MS technology,and the possible mechanism related to anxiety was revealed from the perspective of metabolism.Then using 16S rRNA high-throughput sequencing technology to study the structural characteristics of the intestinal flora of anxious rats,and analyzed the anxiety-related microorganisms.Furthermore,we studied how the structural changes of intestinal flora in anxiety rats affect their metabolic phenotypes,thus revealing the possible physiological mechanism of intestinal flora involved in the onset of anxiety,which is of great significance for the timely diagnosis and effective treatment of anxiety patients in the future.In addition,we use molecular biology and metabonomics to explore the possible and effective way of raw and wine Schisandra chinensis in the treatment of anxiety,in order to provide Chinese medicine ideas for finding safe and effective natural anti anxiety drugs. |
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