| Fibromyalgia syndrome(FM)is a common rheumatic disease.Due to significant individual differences in clinical symptoms,diagnosis is based on subjective evaluations by doctors and patients,and there is a lack of objective diagnostic indicators,which makes early diagnosis difficult.The current treatment methods have limited efficacy.The pathogenesis of FM is not yet fully understood,and there are differences in brain function and gut microbiota in FM patients.However,research is often conducted separately,and systematic research based on the microbiota-gut-brain axis needs to be carried out.Traditional Chinese medicine has achieved good therapeutic effects in treating FM based on syndrome differentiation.Many doctors believe that the disease is mainly caused by the liver,or the seven emotions are not smooth,liver qi stagnation,or liver blood deficiency.At the same time,it involves pathogenesis such as spleen and stomach weakness and insufficient yang qi.Syndrome research has found that ganyu-qizhi syndrome is the main traditional Chinese medicine syndrome type of disease.Research purpose:A meta-analysis was conducted to search for abnormal brain functional connections in resting brain regions of FM patients,and to explore the internal mechanism of changes in the microbiota-gut-brain axis of FM with ganyu-qizhi syndrome,providing a basis for the diagnosis and treatment of FM in traditional Chinese and Western medicine.Part 1:Meta-analysis of resting brain functional connectivity in FM patientsMethod:Using voxel-wise meta-analysis,SDM-PSI software was used to analyze brain functional connectivity abnormalities in resting-state functional magnetic resonance imaging(fMRI)of FM patients.Result:A total of 236 FM patients and 225 control subjects were included in 12 studies with 11 articles.Meta-analysis showed that there were 7 different brain regions.Compared with the control group,the left anterior cingulate gyrus function was enhanced,and the right superior temporal gyrus,the right posterior central gyrus,the right dorsolateral superior frontal gyrus,the right supramarginal gyrus,the right insular lobe,and the left insular lobe function were weakened.After using family-wise error rate(FWER),only the left anterior cingulate gyrus function of FM patients was enhanced(SDM-Z=3.692,P=0.008).Jackknife sensitivity analysis showed that the meta-analysis results were replicable,but there was potential publication bias.Part 2:Clinical Research22 FM patients with ganyu-qizhi syndrome and 20 healthy controls were included.General information about the subjects was collected,and clinical symptoms such as pain,anxiety,depression,fatigue,and sleep were evaluated using a scale.The following research was conducted.Study 1:Differences in brain structure and function in FM patients with ganyu-qizhi syndromeMethod:A 3.0T magnetic resonance scanner was used to collect brain structure and resting-state functional scanning data.1.Based on the voxel-based morphological analysis method(VBM),SPM12 software VBM8 toolbox was used for data processing to obtain brain regions of gray matter volume changes in FM patients with ganyu-qizhi syndrome and control group.2.On the MATLAB 2018b platform,SPM12 software and the DPABI toolkit were used to preprocess fMRI data.①Low-frequency oscillation amplitude(ALFF)analysis was used to reflect the characteristics of local neural activity in the brain,and the brain regions with differences in ALFF values between FM patients with ganyu-qizhi syndrome and the control group were compared.②Based on seed point functional connectivity analysis,different brain regions in the meta-analysis were selected:the left anterior cingulate gyrus and the right superior temporal gyrus as seed points,and the whole brain functional connectivity mapping map was calculated to statistically analyze the brain regions with differences in functional connectivity between the ganyu-qizhi syndrome FM and the control group;③Independent Component Analysis(ICA)used GIFT software to evaluate the optimal decomposition score,matched with 14 resting-state network components,and statistically analyzed the brain networks of FM patients with ganyu-qizhi syndrome with abnormal functional connectivity.3.SPSS 26.0 software was used to analyze the general information and clinical evaluation scale of the subjects.The revised version of the Fibromyalgia Syndrome Impact Questionnaire for FM patients,NRS score,fatigue scale,anxiety self-evaluation scale,depression self-evaluation scale.Pittsburgh Sleep Quality Index score,abnormal gray matter volume,and ALFF value were analyzed for correlation.Research results:1.There is no significant difference in gender,age,education years,and BMI between FM patients with ganyu-qizhi syndrome and the healthy control group.FM patients with ganyu-qizhi syndrome have mild anxiety and depression,high fatigue scale scores.and significant physical fatigue.2.VBM analysis After GRF correction(P<0.05),compared with the healthy control group,there were 8 brain areas with reduced gray matter volume in the FM patients with liver stagnation and qi stagnation syndrome,respectively the left middle frontal gyrus,the right middle cingulate gyrus,the right paracentral lobule,the left middle temporal gyrus,the right superior parietal gyrus,the left anterior cuneiform lobe,the left superior occipital gyrus,and the right insular lobe.The two brain areas with increased gray matter volume were the right superior temporal gyrus and the left thalamus.There was no significant difference(P>0.05)in the correlation analysis between the differential gray matter volume in the whole brain of FM patients with ganyu-qizhi syndrome,as well as the duration of the disease and the scores of various scales.3.After FDR correction(P<0.05),compared with the healthy control,the ALFF values in two brain regions of the FM patients with ganyuqizhi syndrome increased,namely,the right precentral gyrus and the left subfrontal orbital region,while the ALFF values in six brain regions decreased,namely,the left medial superior frontal gyrus,the left middle occipital gyrus,the left dorsolateral superior frontal gyrus,the left parietal inferior marginal angle,the right posterior central gyrus,and the left middle frontal gyrus.There was no significant difference(P>0.05)in the correlation analysis between the differential ALFF value and the duration of illness and the scores of various scales.4.With the left anterior cingulate gyrus as the seed point,compared with the healthy control,there were three brain areas with enhanced connectivity in the FM patients with liver stagnation and qi stagnation syndrome,namely,the left posterior cerebellar lobe,the left lenticular nucleus,and the right precentral gyrus.No brain area with weakened functional connectivity was found.With the right superior temporal gyrus as the seed point,compared with the healthy controls,the patients with ganyu-qizhi syndrome FM had three brain areas with enhanced connectivity,which were divided into the left precentral gyrus,the left occipital lobe,and the left middle frontal gyrus.The four brain areas with weakened functional connectivity were the right middle temporal gyrus,the right middle frontal gyrus,the left angular gyrus,and the right anterior cuneiform lobe.5.ICA analysis showed that 32 components were matched with 14 resting networks,including the dorsal default mode network,the ventral default mode network,the highlighted network,and the basal ganglia network.Statistical analysis did not find that there was a significant difference in the brain network between the patients with ganyu-qizhi syndrome and the control group(P>0.05).6.Meta-analysis of fMRI in resting-state in patients with abnormal brain areas of FM of liver stagnation and qi stagnation syndrome and those with FM of undifferentiated syndrome type found abnormalities in the right posterior central gyrus and the dorsolateral superior frontal gyrus.Study 2:Changes in the microbiota gut brain axis in FM patients with liver stagnation and qi stagnation syndromeMethod:1.Collect feces from subjects and use the 16S rDNA sequencing method to obtain species information of variable regions of intestinal bacteria V3-V4;Use the ITS sequencing method to obtain species information of intestinal fungi.Use QIIME software for data preprocessing,obtain species annotation information,and conduct alpha diversity,beta diversity analysis,and inter-group community difference analysis.2.Collect the serum of the subjects and perform untargeted metabolomics detection of serum metabolites using liquid chromatography-mass spectrometry(LC-MS).The data was preprocessed using Compound Discover(CD3.1)software and R(v3.1.3)XCMS package.Multiple statistical analysis was conducted to screen differential metabolites in FM patients with ganyu-qizhi syndrome,and enrichment analysis was performed.3.Network analysis integrates clinical data,gut microbiota,serum metabolites,and differences in gray matter volume of FM patients with liver stagnation and qi stagnation syndrome,as well as functional magnetic resonance brain connectivity data.Research results:1.The 16S rDNA analysis of intestinal bacterial flora showed that the dominant bacteria and species were the same in the patients with ganyu-qizhi syndrome FM and the control group,and the difference microorganisms between the LEfSe analysis groups.In the FM patients with ganyu-qizhi syndrome,the abundance of Akkermansia,Enterococcus,Lactococcus increased,and the abundance of Eubacterium,Lachnospira,Ruminococcus decreased.Analysis of bacterial function prediction shows that the top 10 functional pathways mainly involve metabolism,genetic information processing,and cellular processes.There was no significant difference in Alpha diversity and Beta diversity.2.ITS identification of intestinal fungi showed that FM patients with ganyu-qizhi syndrome had the same dominant bacterial phylum as the control group,but there were differences in the dominant bacterial genera.The characteristic fungal microorganisms among the groups were analyzed by the LEfSe method.The abundance of Flammulina and Angulomyces in FM patients with ganyu-qizhi syndrome increased,while the abundance of Alternaria,Lodderomyces,Saccharomycopsis,and Dolichousnea decreased.There was no significant difference in Alpha diversity and Beta diversity.3.Untargeted serum metabolomics analysis screened 57 differential metabolites(VIP>1 and P<0.05),and further screened 13 differential metabolites based on multiple differences.Topological analysis and enrichment analysis identified five significant metabolic pathways,with varying influences ranging from high to low,including tryptophan metabolism.arginine and proline metabolism,steroid hormone biosynthesis,phenylalanine metabolism,and glutathione metabolism.4.Correlation analysis showed that there was a wide connection between intestinal bacteria and fungi.Correlation analysis between intestinal bacteria and serum differential metabolites showed that Akkermansia was positively correlated with jasmonic acid.N-lactylphenylalanine and benzoyl acid,while negatively correlated with Lmethionine and L-pyroglutamine.The Eubacterium is positively correlated with indole-3-acetic acid and L-methionine,while negatively correlated with levothyroxine and maslinic acid.Ruminococcus are positively correlated with acetylcholine,DL-lysine,L-citrulline,while negatively correlated with methyl propionate and levothyroxine.Enterococcus was positively correlated with levothyroxine.The Alternaria in fungi was negatively correlated with DL-lysine,isoliquiritigenin,and N-lactylphenylalanine.Dolichousnea was negatively correlated with methyl propionate and benzoic acid.Flammulina was negatively correlated with 17 αHydroxyprogesterone,acetylcholine,creatinine and cystine.Lodderomyces was negatively correlated with Kynurenic acid.Saccharomycopsis was positively correlated with 17 αHydroxyl ketone and corticosterone,while negatively correlated with methyl propionate.5.The analysis of the microbiota gut brain axis network showed that there was a closer correlation between symptom evaluation scales,between gut fungi and bacteria,and between serum metabolites and gut microbiota.The difference in whole-brain VBM was only negatively correlated with the fungal genus Dolichomonas,while the difference in whole-brain ALFF was not significantly correlated with other indicators.Conclusion:1.A voxel-based meta-analysis was conducted to organize the abnormal functional brain regions of resting fMRI in FM patients,involving multiple brain regions such as pain perception,pain processing,cognition,and emotional regulation.2.There are abnormalities in the brain structure and function of FM patients with ganyu-qizhi syndrome,which are similar to the results of meta-analysis without syndrome differentiation.There are also unique abnormal brain functional regions in the brain.3.Multiomics analysis shows that there are multiple potential biomarkers in the gut bacteria,fungi,and serum metabolites of FM patients with liver stagnation and qi stagnation syndrome.The gut characteristic microorganisms are closely related to serum differential metabolites,and the same metabolic pathway involved may be the biological change pathway of the gut-brain axis in FM patients with liver stagnation and qi stagnation syndrome,which is related to the traditional Chinese medicine theory of integrating form and spirit,and the influence of liver stagnation and qi stagnation on the large intestine conduction function. |
PDF Full Text Request