| Objective:1.Evaluate the clinical efficacy of Chaihu-jia-Longgu-Muli decoction in treating patients with persistent postural and perceptual dizziness accompanied by anxiety through clinical studies.2.Utilize ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry(UPLC-Q-TOF/MS)to isolate and identify the active components of Chaihujia-Longgu-Muli decoction.3.Predict the mechanism of action of Chaihu-jia-Longgu-Muli decoction in treating vertigo accompanied by anxiety through targeted network pharmacological research methods.4.Validate the results of network pharmacological studies through animal experiments,observe the behavior,neurotransmitter levels,and the expression of proteins in the BDNF/Trk B/CREB signaling pathway in rats with a model of vertigo and anxiety,to clarify the mechanism of action of Chaihu-jia-Longgu-Muli decoction in treating vertigo and anxiety.Methods:1.A total of 120 patients with persistent postural and perceptual dizziness accompanied by anxiety who visited the Dizziness and Balance Disorders Diagnosis and Treatment Center at the First Clinical Hospital of Jilin Academy of Traditional Chinese Medicine from January 2023 to December 2023 were randomly divided into a control group and an experimental group,with 60 patients in each group.There were no statistically significant differences in baseline characteristics such as age,gender,and duration of illness between the two groups.The control group received conventional Western medical treatment,including medication and vestibular rehabilitation therapy.The experimental group received modified Chaihu-jia-Longgu-Muli decoction in addition to the treatment given to the control group,with both groups undergoing continuous treatment for 6 weeks.The improvement in Dizziness Handicap Inventory scores,Dizziness Visual Analog Scale scores,Hamilton Anxiety Scale scores,and Traditional Chinese Medicine syndrome scores at different time points was observed to evaluate the efficacy of Chaihu-jia-Longgu-Muli decoction in treating dizziness accompanied by anxiety.2.A chemical composition database of Chaihu-jia-Longgu-Muli decoction was established by searching databases and literature.Ultra-performance liquid chromatographyquadrupole time-of-flight mass spectrometry was used to perform positive and negative ion scans on the decoction’s decoction,and the UNIFI software was employed for automatic retrieval and matching.Standard substances and literature were utilized to identify the in vitro components of Chaihu-jia-Longgu-Muli decoction.Based on the in vitro components,plasma samples containing the decoction’s components were collected from rats at different time points after oral administration.By comparing these samples with blank plasma,the prototype components of Chaihu-jia-Longgu-Muli decoction were screened.3.Target prediction was conducted using Swiss Target Prediction on the prototype components in the blood.Target information related to dizziness and anxiety was obtained from Gencards,Drugbank,and OMIM databases,and the intersection targets between drugs and diseases were obtained using the Venny 2.1 platform.A target network diagram of the Chaihu-jia-Longgu-Muli decoction’s prototype components in the blood for treating dizziness accompanied by anxiety was constructed,depicting protein interaction network effects.The common targets were inputted into the DAVID database for GO and KEGG enrichment analysis to predict the mechanism of action of Chaihu-jia-Longgu-Muli decoction in treating dizziness accompanied by anxiety.4.Model rats with vertigo with anxiety were established and divided into model group,Chaihu-jia-Longgu-Muli decoction and paroxetine group.After continuous treatment for 7 days,behavioral,neurotransmitter content and BDNF/Trk B/CREB signaling pathway protein expression of rats in different groups were detected,so as to verify the mechanism of action of Chaihu-jia-Longgu-Muli decoction in the treatment of vertigo and anxiety.4.A rat model of dizziness accompanied by anxiety was established and divided into a model group,a Chaihu-jia-Longgu-Muli decoction group,and a paroxetine group.The rats were treated continuously for 7 days,and their behavioral characteristics,neurotransmitter levels,and the expression of BDNF/Trk B/CREB signaling pathway proteins were measured to preliminarily elucidate the mechanism of action of Chaihu-jia-Longgu-Muli decoction in treating dizziness accompanied by anxiety.Results:1.This study included a total of 120 patients,11 of whom dropped out,5 in the experimental group and 6 in the control group.(1)Comparison of baseline data between the two groups: The patients in both groups were comparable in terms of gender,age,duration of illness,Dizziness Handicap Inventory(DHI)score,Hamilton Anxiety Rating Scale(HAMA)score,Vertigo Visual Analog Scale(VAS)score,and Traditional Chinese Medicine(TCM)syndrome score.(2)Comparison of treatment effect between the two groups: After 2and 6 weeks of treatment,the total score of DHI,DHI-P,DHI-F and DHI-E index of patients in the two groups were lower than before treatment.At week 2,the total score of DHI,DHIP,DHI-F and DHI-E index of experimental group were decreased compared with control group,but there was no statistical difference(P > 0.05).At the 6th week,the total score of DHI,DHI-P,DHI-F and DHI-E index in experimental groups were significantly lower than those in control group,with statistical differences between groups(P < 0.05).The results of repeated measurement variance showed that there were statistically significant differences in total DHI,intra-group and interaction(P < 0.05),statistically significant differences in DHIP index within groups(P < 0.05),but no statistically significant differences in inter-group and interaction(P > 0.05).DHI-F index showed statistically significant differences within groups and interaction(P < 0.05),but no statistically significant differences between groups(P > 0.05),DHI-E index showed statistically significant differences within groups(P <0.05),and no statistically significant differences between groups and interaction effects(P >0.05).In terms of improving the vertigo VAS score,the vertigo VAS score of 2,4 and 6weeks after treatment in both groups was decreased compared with that before treatment,and the VAS score of the experimental group was significantly lower than that of the control group at the 2nd and 4th week,with statistical significance(P < 0.05);at the 6th week,there was no statistical difference between the two groups(P > 0.05).The results of repeated measurement variance showed that the differences between groups,within groups and across VAS ratings were statistically significant(P < 0.05).HAMA score showed that both groups of patients were lower than before treatment.Inter-group comparison showed that HAMA score of the experimental group was lower than that of the control group at week 2,but the difference between the two groups was not significant,without statistical significance.At the 6th week,HAMA scores decreased significantly between the two groups(P < 0.05).TCM syndrome scores and curative effects of the experimental group were better than those of the control group as a whole,with statistical significance(P <0.05).Repeated measurement variance results showed that there were statistically significant differences between groups and within groups in HAMA evaluation(P < 0.05),while interaction differences were not statistically significant(P > 0.05).(3)Occurrence of adverse reactions: During the treatment of the two groups of patients,there was one case of sudden acute vestibular disease in the experimental group,which was not related to the treatment plan by analysis.(4)Comparison of follow-up indicators: 1 month after treatment,the total score of DHI,VAS score,HAMA and TCM syndrome score of the experimental group were lower than those of the control group,and the differences between the two groups were statistically significant(P < 0.05).2.Through UPLC-Q-TOF/MS technology combined with UNIFI,102 active ingredients were identified,including 38 saponins,32 flavonoids,12 polyphenols,9anthraquinones and 11 other compounds.21 of the active components can be absorbed into the blood.3.The pharmacological study of blood entry network showed that 282 targets could be regulated by 21 blood entry components of Chaihu-jia-Longgu-Muli decoction,and 4047disease-related targets were obtained,81 key targets were obtained by intersection of the two.PPI network analysis showed that the key targets of Chaihu-jia-Longgu-Muli decoction in treating vertigo and anxiety were mainly VEGFA,DRD2,Caspase-3,EGFR,etc.GO and KEGG mainly enrich neuronal synapses,neurotransmitter regulation,neuroligand-receptor interactions,5-hydroxytryptamine synapses,and neurotrophic factor signaling pathways.4.In this study,unilateral labyrinth destruction combined with anxiety index was used to establish a vertigo and anxiety model.Through Petrosini reporting method,rotary rod experiment and elevated cross maze behavioral experiment,it was verified that the model rats could show postural imbalance,decreased motor coordination ability and anxiety behavioral characteristics due to vestibular damage.(1)Behavioral evaluation.After intragastric treatment,the unbalanced behavior of rats in sham operation group,Chaihu jia Longgu Muli decoction group and paroxetine group disappeared completely,while some rats in model group still had behavioral imbalance symptoms,but the difference was not statistically significant compared with the other three groups.The rotating rod experiment showed that the average latency of rats staying on the rotating rod increased in all groups,and the average latency of the other three groups was roughly the same as that of the sham operation,and basically returned to the normal level,with no statistical significance between the groups.The elevated cross maze showed that compared with the model group,the time of entering the open arm was longer in the Chaihu-jia-Longgu-Muli decoction group and the paroxetine group(P<0.05),and the number of entering the open arm in the paroxetine group was higher than that in the model group(P<0.05).Although the number of entering the open arm in the Chaihu-jia-Longgu-Muli decoction was higher than that in the model group,However,there was no statistical difference between the groups(P=0.07),and the time of entering the closed arm and the anxiety index of the Chaihu-jia-Longgu-Muli decoction group and the paroxetine group decreased.(2)Changes in neurotransmitter content.Compared with sham operation,NE and 5-HT contents in model group were significantly increased(P<0.05),and DA level was higher than that in sham operation group,but there was no statistical difference between the two groups(P=0.135).Compared with the model group,the levels of NE and 5-HT in both the Chaihu jia Longgu Muli decoction group and the paroxetine group were decreased(P<0.05),and the decrease level of NE in the Chaihu jia Longgu Muli decoction group was more significant than that in the model group(P< 0.01).On the 7th day,there was no statistical difference in DA content among all groups,and the DA content in Chaihu-jia-Longgu-Muli decoction group decreased the most.(3)Westernblot results.The expression levels of BDNF,Trk B and CREB were similar between sham operation and model group,but the difference was not statistically significant.Compared with model group,the expressions of BDNF,Trk B and CREB in Chaihu-jiaLonggu-Muli decoction group were increased,and the differences were statistically significant(P < 0.05).Conclusion:1.Chaihu-jia-Longgu-Muli decoction has definite curative effect in the treatment of PPPD with anxiety,which can reduce the symptoms and severity of vertigo in patients with PPPD accompanied by anxiety,and improve the anxiety state,which is worthy of clinical promotion and application.2.A total of 102 kinds of active ingredients were identified from the Chaihu jia Longgu Muli decoction,including saponins and flavonoids.Among them,21 active components can be absorbed by the blood,which may be the main material basis for the efficacy of Chaihu jia Longgu Muli decoction.3.Network pharmacological studies showed that there were 81 key targets of 21 bloodentering components in the decoction,and GO and KEGG mainly enriched neuronal synapses,neurotransmitter regulation and neurotrophic factor signaling pathways,indicating that the Chaihu jia Longgu Muli decoction played a therapeutic role through multicomponents,multi-targets and multi-pathways.4.Chaihu jia Longgu Muli decoction may promote the repair of cells in the vestibular nucleus of the injured side,accelerate vestibular compensation and relieve the symptom of imbalance by up-regulating the BDNF/Trk B/CREB signaling pathway.At the same time,it can reduce the content of neurotransmitters in the brain and play an anti-anxiety role. 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