| Objective: This study aimed to analyze the influence of harmonizing stomach to tranquilize mind acupuncture on the intestinal microbiota in patients with insomnia due to spleen-stomach disharmony syndrome through 16 S r DNA high-throughput sequencing technology.And explore the mechanism of its treatment efficacy on insomnia due to spleen-stomach disharmony syndrome.Methods: 1.Twenty-four eligible patients with insomnia due to spleen-stomach disharmony syndrome were randomly divided into two groups: 12 in treatment group and 12 in control group.Those in the treatment group received harmonizing stomach to tranquilize mind acupuncture,while those in the control group received sham acupuncture.All patients received 5 acupuncture treatment sessions per week for 4 consecutive weeks.Besides,10 healthy volunteers were be recruited as healthy group.2.the Pittsburgh Sleep Quality Index(PSQI);the Gastrointestinal Symptom Rating Scale(GSRS);Self-Rating Anxiety Scale(SAS),Self-Rating Depression Scale(SDS),and State-Trait Anxiety Inventory(STAI)were used to compared the clinical efficacy of treatment group and control group;16S r DNA high-throughput sequencing was used to detect differences in intestinal microbiota structure and diversity;high-performance liquid chromatography-mass spectrometry was used to detect the g-aminobutyric acid(GABA)content and glutamic acid(Glu)content.The treatment group and the control group were collected the above clinical evaluation indicators and fecal samples at the time of enrollment and after the end of treatment,and the healthy group were collected fecal samples at the time of enrollment.Results: 1.A total of 33 subjects completed the study: 10 in the healthy group;23 in the insomnia group,including 12 in the treatment group and 11 in the control group.2.Comparison of clinical efficacy: the PSQI score and GSRS score of the treatment group were significant lower than those in the control group(P<0.05);There were no significant differences in the scores of SAS,SDS and STAI between the two groups(P>0.05).3.Comparison of intestinal microbiota:(1)Comparison between healthy group and insomnia group: The relative abundance of Fusobacteria in the insomnia group was significantly lower than that in the healthy group(P<0.05).LEf Se analysis(LDA threshold 2)showed that compared with insomnia group,Betaproteobacteria,Burkholderiales,Sutterella,Alcaligenaceae,Aeromonadaceae,Aeromonas,Pseudomonas,and Pseudomonadaceae were significantly enriched in the healthy group.(2)Comparison between the control group and the treatment group: Before treatment,the relative abundance of Tenericutes in the treatment group was significantly lower than that in the control group(P<0.05),and the Observed OTU and Faith PD index were significantly lower than those in the control group(P<0.01);After treatment,there was no significant difference in species composition and Alpha diversity indexes between the two groups(P>0.05).After treatment,the relative abundance of Ruminococcus and the Faith PD index were significantly increased in the treatment group(P<0.05);there was no significant difference in species composition and Alpha diversity index before and after treatment in the control group(P>0.05).(3)Statistical analysis of intestinal flora and clinical efficacy: SMB53,Blautia,and [Ruminococcus] were positively correlated with PSQI total score;Paraprevotella,Desulfovibrio,Faecalibacterium,Comamonas,and Haemophilus were negatively correlated with PSQI total score.SMB53 was positively correlated with GSRS total score;Sutterella,Pseudomonas,and Faecalibacterium were negatively correlated with GSRS total score.4.Comparison of GABA and Glu content: Glu content in the insomnia group was significantly higher than that in the healthy group(P<0.05).There was no significant difference in GABA and Glu content levels between inter or inner group comparison(P>0.05).Conclusions:1.The harmonizing stomach to tranquilize mind acupuncture can improve the sleep quality of patients with insomnia due to spleen-stomach disharmony syndrome and improve their gastrointestinal symptoms.2.Structural disorders of the intestinal microbiota in patients with insomnia due to spleen-stomach disharmony syndrome.With the change of intestinal microbiota,the metabolite Glu content was significantly higher than that of the healthy group.3.And the mechanism of improving the sleep quality by harmonizing stomach to tranquilize mind acupuncture may be related to the improvement of intestinal microbiota diversity,but has no obvious correlation with the metabolites Glu and GABA. |
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