Effects Of Methadone Maintenance Treatment And Compulsory Isolation Detoxification On Gut Microbial Diversity

Objective: This study used drug users and healthy people as controls to explore the effects of Methadone Maintenance Treatment（MMT）and compulsory isolation detoxification treatment on the composition and diversity of gut microbes,as two widely used treatment methods for drug detoxification.It is expected to provide scientific data for the improvement of possible gut-related diseases in drug users,methadone maintenance treatment patients,and compulsory isolation detoxification populations,and to provide valuable information for promoting different detoxification treatment methods for drug users’ physical and mental health recovery and smooth return to society.Methods: The study selected Honghe Prefecture,Yunnan Province as the study site,and adopted strict inclusion and exclusion criteria.Finally,the demographic basic information data and dietary frequency questionnaire of 99 survey subjects [divided into four groups: compulsory isolation detoxification group（CD）;methadone maintenance treatment patients group（MP）;drug use control group（DU）;healthy control group（HC）] were analyzed,and fecal samples were collected to extract gut microbial genomic DNA,and 16 S r DNA sequencing analysis was performed.Then we analyzed the dietary nutrition intake status of each group,the difference of gut microbial diversity,the potential relationship between diet and the diversity of gut microbes,and explored the main driving factors that affect the gut microbes of each group.Results:（1）General demographic basic information data in CD,MP,DU,and HC showed significant differences in gender（X~2 = 15.469,P = 0.001）,age（X~2 =33.957,P < 0.001）and education level（X~2 = 32.802,P < 0.001）.Metadata regarding drug abuse behavior among CD,MP,and DU study subjects showed significant differences among these three groups,notably in the duration of drug abuse（in months）（P < 0.001;Kruskal-Wallis rank sum test）,drug type（X~2 = 15.340,P = 0.004）,and frequency of forced cessation（X~2 = 41.892,P < 0.001）,while drug abuse manner in the three groups was no statistical difference（X~2 = 3.257,P = 0.516）.（2）Principal component analysis（PCA）of dietary structure showed that the dietary structure of MP group was quite different compared with other groups.The four groups of dietary nutrition intake data showed that the MP has the characteristics of higher dietary intake and energy intake.In particular,the intake of fruits,milk,meat,and nuts were higher in MP than that in CD and DU（P ≤ 0.001-0.025）.Proteinand carbohydrate-based energy,and dietary fiber intake in the MP group were the highest compared with other groups（P ≤ 0.001-0.046）.（3）The alpha diversity data of the gut microbial communities among four groups showed that differences in the richness and abundance of OTUs were non-significant among groups in comparisons of observed species,Chao1,Abundance-based coverage estimator（ACE）,and Shannon and Simpson indices,although differences in the coverage index among the four groups were significant（P = 0.035）.（4）Principal coordinate analysis（PCo A）analysis based on Jaccard distance showed that HC and CD were separated along axis 2,with some overlap.However,microbiota of DU and MP were highly similar.Linear discriminant analysis（LDA）effect size（LEf Se）showed that Actinobacteria,Streptococcaceae,Lactobacillaceae,Bifidobacteriaceae,Enterobacteriaceae,Lactobacillus,Streptococcus,Veillonella,Bifidobacterium,Intestinibacter and Fusicatenibacter were significantly enriched in MP group（LDA = 2.824-4.440,P ≤ 0.001-0.014）.The gut microbes of DU were more abundant in Peptostreptococcaceae,Succinivibrionaceae,Coriobacteriaceae,Ruminococcus,Roseburia,and Collinsella and Succinivibrio（LDA = 3.043-4.366,P≤ 0.001-0.049）.（5）The analysis of variance（ANOVA）of gut microbial species abundance between groups showed that there was no significant difference between CD and HC in the levels of phylum,class,order and family.The proportions of Bifidobacteriaceae,Lactobacillaceae,Bifidobacterium and Lactobacillus were the highest in MP than others（P ≤ 0.001-0.024）.MP showed a higher proportion of Actinobacteria compared with CD and HC（P = 0.005-0.017）.Similarly,Streptococcaceae,Streptococcus and Fusicatenibacter were found in higher levels in MP than in CD and DU（P =0.021-0.043）.In addition,The gut microbiota in DU were significantly enriched for Firmicutes and Ruminococcaceae compared with that in CD（P = 0.004-0.021）.Among the four subject groups,DU also showed the highest proportional abundance of Ruminococcus（P = 0.001-0.038）.（6）The Pearson correlation analysis of gut microbes and diet showed that specific OTUs are significantly correlated with dietary intake.Energy from carbohydrates was positively correlated with the presence of Lachnospira,Anaerostipes,Fusicatenibacter,Cellulosilyticum and Atopobium（r = 0.306-0.373,P ≤0.001-0.002）.The intake of dietary fiber was positively correlated with the abundance of Papillibacter（r = 0.380,P < 0.001）.The abundance of Cyanobacteria,Betaproteobacteria,Burkholderiales,Bifidobacteriales,Actinomycetales,Sutterellaceae,Lactobacillaceae,Bifidobacteriaceae,Chloroplast,Microbacteriaceae,Enterobacteriaceae,Lactobacillus,Bifidobacterium,Coprobacillus,Streptophyta,Hungatella,Okibacterium,Howardella,Enterococcus,Ezakiella,and Methanosphaera were positively correlated with milk intake（r = 0.301-0.436,P ≤0.001-0.002）.Conclusion:（1）The high beta diversity of gut microbes in MP may be driven by dietary changes caused by drugs（methadone intake leads to an increase in orexin A,which in turn affects the diet and causes the enrichment of Lactobacillus,Bifidobacterium and Fusicatenibacter in the gut）,these changes indicate that methadone maintenance treatment may affect gut health through dietary regulation.（2）CD and HC hold higher similarity of gut microbial diversity,which indicates that compulsory isolation treatment may be beneficial to improve the gut health of drug users.（3）The high abundance of Ruminococcus in the gut microbes associated with the increased risk of memory and cognitive impairment was lower in MP and CD than DU,suggesting that both drug detoxification methods may reduce the risk of memory and cognitive impairment in drug users.（4）Both methadone maintenance treatment and compulsory isolation treatment are beneficial to gut health to a certain extent.