| Objective:The correlation between intestinal microecology and common chronic liver disease damp-heat syndrome is explained from the point of TCM theory that different diseases with a same syndrome,which provides scientific basis for the objective study of TCM syndromes.Methods:A cross-sectional clinical epidemiological study was conducted to study patients with chronic hepatitis B(CHB)and non-alcoholic fatty liver(NAFLD)dampness-heat syndrome.Non-damp and heat patients with the above-mentioned diseases were used as the control group,and healthy volunteers were used as the control group.The information of TCM clinical diagnosis and Western medicine diagnosis of the above-mentioned volunteers were collected,and stool samples were collected for biodiversity sequencing in the V3-V4 region of 16 S rRNA(16S ribosomal RNA)gene.On the basis of expert evaluation of syndrome typicality,the clinical characteristics of patients with dampness-heat syndrome under two disease backgrounds were firstly analyzed.Secondly,microbial diversity data were analyzed.Alpha diversity analysis was used to compare species richness and diversity.Principal component analysis(PCA),PCOA(Principal Co-ordinates Analysis)and other analyses were used,combined with ANOSIM/Adonis inter-group difference test.To assess the similarities or differences in overall community structure among different samples/groups;Based on the species annotation results,the species composition of each group was analyzed to obtain the composition and abundance changes of dominant species in each group.Species with significant differences between groups were obtained by assessing the significance of species abundance differences.The generality and individuality of intestinal microecology of CHB and NAFLD patients with typical dampness-heat syndrome were summarized.On this basis,a syndrome diagnosis model for common chronic liver diseases with dampness-heat syndrome was established.The receiver operating characteristic curve(receiver operating characteristic curve)was applied.ROC)to evaluate the diagnostic efficiency of the model.Results :(1)A total of 405 volunteers were recruited in this study,including 127 patients with CHB(77 patients with dampness-heat syndrome and 50 patients with non-dampness-heat syndrome),183 patients with NAFLD(109 patients with dampness-heat syndrome and 74 patients without dampness-heat syndrome),and 95 healthy volunteers.The level of chronic liver disease patients with dampness-heat syndrome was significantly higher than that of patients with non-dampness-heat syndrome,and the frequency of mouth bitterness,moss yellow,moss greasy,tongue red and pulse slippery number in the four syndromes was significantly higher than that of patients with non-dampness-heat syndrome(P < 0.05).(2)Comparing the effects of diseases or syndrome groups on intestinal flora,the analysis of 405 volunteers showed that the intestinal microbes could not be "naturally" differentiated according to different diseases or syndrome groups by unsupervised analysis method.However,PLS-DA(Partial Least Squares Discriminant Analysis)could distinguish intestinal bacteria from healthy volunteers,CHB patients and NAFLD patients.Using the same method according to the syndromes grouping,the patients with damp-heat syndrome and non-damp-heat syndrome could not be significantly separated,and the damp-heat syndrome groups of the two diseases did not show the trend of aggregation,suggesting that the difference of flora between different diseases is greater than the difference between syndromes.The potential interference of syndrome typical degree difference to the analysis of characteristic microflora of syndrome was considered.Therefore,follow-up studies were conducted to analyze the commonness and individuality of changes in intestinal flora of damp and heat syndrome in common chronic liver diseases by taking typical dampness-heat syndrome patients and non-dampness-heat syndrome patients without the secondary syndrome of damp and heat as objects.(3)Analysis of the commonality and personality of changes in intestinal flora diversity in CHB and NAFLD patients with typical damp-heat syndrome showed that there was no statistical significance in the diversity of intestinal flora Alpha between CHB and NAFLD patients with typical damp-heat syndrome and patients without damp-heat syndrome.However,PLS-DA analysis of intestinal bacteria between typical and non-damp-heat patients could be clearly distinguished,suggesting that there is a material basis for the difference of bacterial flora between typical and non-damp-heat patients.Species difference analysis showed that at the phylum level,there was no statistical difference between CHB and NAFLD patients with typical and non-damp-heat syndromes.At the generic level,compared with CHB patients without damp-heat syndrome,Dorea,Dialister,Ruminococcus＿Gauvreaui＿Group,Adlercreutzia,Unclassified＿P＿Firmicutes,Unclassified＿C＿Clostridi of typical CHB patients with dampheat syndrome A,the abundance of Unclassified＿F＿Veillonellaceae,UCG-003 and Unclassified＿C＿＿Bacilli were significantly lower(P<0.05);The abundance of Fenollaria and Bacillus were significantly increased(P<0.05).The abundance of Bautia,Abiotrophia and Pseudoproionibacterium in typical NAFLD patients were significantly decreased(P<0.05)compared with those in non-dampness and heat syndrome patients.The abundance of Non＿F＿Norank＿O＿RF39,Shuttleworthia,Atopobium and Alloscardovia were significantly increased(P<0.05).722 OTUs(Operational Taxonomic Units)with common expression trends between CHB and NAFLD patients with typical damp-heat syndrome and non-damp-heat syndrome,mainly involving 11 taxa and 191 genera,and 127 genera with the same trend.The genera with common expression trends were found to be mainly involved in the metabolism of arginine and proline,biosynthesis of streptomycin,degradation of aromatic compounds,type 2 diabetes,human papillomavirus infection and viral carcinogenesis.(4)Bacteria with common expression trend and statistical P value <0.1 between CHB and NAFLD patients with damp-heat syndrome and non-damp-heat syndrome were screened.Combined with the common difference of clinical index between patients with damp-heat syndrome and non-damp-heat syndrome and the non-subjective symptom of traditional Chinese medicine,the discriminant model of damp-heat syndrome of chronic liver disease was established.The Area Under Curve(AUC)of ROC of CHB patients with typical and non-dampness-heat syndrome was 0.872.AUC of NAFLD patients with typical dampness-heat syndrome and non-dampness-heat syndrome was 0.851.A total of405 CHB and NAFLD patients were cross-validated.The AUC of CHB patients with damp-heat syndrome was 0.752.The AUC of patients with dampness-heat syndrome in NAFLD was 0.712.Conclusion:1.The difference of intestinal flora between diseases was greater than that between syndromes2.At the level of different diseases and syndrome,we found that there were 722 enterobacteria with common expression trend in CHB and NAFLD patients with dampness-heat syndrome at the OTU level and 127 at the generic level.These enterobacteria may be involved in methane metabolism and β-alanine metabolism pathway.3.According to the regression equation constructed by Eubacterium,Atobacterium,DBIL and mossy yellow,the AUC of cross-validation for CHB and NAFLD patients with damp heat syndrome were 0.752 and 0.712,respectively,which need to be further verified. |
PDF Full Text Request