Preliminary Study On The Relationship Between Changes Of Oral Metabolites And HSV-1,Bacterial And Fungal Loads

Objective:After HIV infection,there are some oral lesions associated with the microbial infection,including oral candidiasis and herpetic stomatitis.The analysis of the relationship between metabolites and microorganisms in the oral cavity of patients with HIV infection is helpful to strengthen the understanding of the role of microorganisms in oral lesions.In this study,through case-control design,LC-MS metabonomics and qPCR techniques were used to explore the relationship between differential metabolites and main microorganisms HSV-1,bacteria and fungi in oral fluid of HIV infected and healthy people.Methods:The HIV infected group,was the HIV infected population without HAART,with 20 cases.the healthy control group,was population who were negative for HIV antibody,with 19 cases.We collected oral gargle,then the differential metabolite information of the two groups was obtained by metabolite extraction,LC-MS non-target metabonomics detection,qualitative and semi-quantitative analysis and bioinformatics analysis.On the basis of the DNA extracted from gargle,qPCR was adopted to quantify HSV-1,bacteria and fungi in samples.Finally,the correlation between differential metabolites and the microbial load was analyzedResults:(1)Microbial load(x±s):The microbial loads HSV-1 of Group A and Group R were 4.4427±2.0815 and 3.3483±3.0685 respectively,the bacteria were 3.9113±2.2416 and 4.1422±1.9808 respectively,and the fungi were 4.3957±1.7305 and 4.7994±1.9946 respectively.Statistical analysis showed that the mean value HSV-1 of the logarithm of the number of copies per microliter was relatively high in Group A,and the difference was statistically significant(P<0.05).The mean values of bacteria and fungi were relatively high in Group R,and the difference was not statistically significant(P=0.911 and P=0.448).(2)Differential metabolite information:a total of 228 metabolites were detected in the positive ion mode,and 33 differential metabolites were detected in the two groups.In the negative ion mode,98 metabolites were detected in the two groups,and there were 15 differential metabolites between the two groups.(3)Correlation analysis:①In positive ion mode,the number of differential metabolites associated with HSV-1,bacteria and fungi were 4,13 and 28,respectively.②In negative ion mode,the number of differential metabolites related to HSV-1,bacteria and fungi were 1,0 and 6,respectively.(4)Hierarchical cluster analysis of differential metabolites related to microbial load:①In positive ion mode,the amount of most dipeptides increased and the amount of choline decreased in Group A.②In the negative ion mode,it was found that the amounts of 2’-Deoxyuridine,phosphatidylcholine and heptadecanoic acid in Group A were significantly higher than those in other substances,suggesting that the metabolism of these substances in Group A was abnormal.(5)KEGG analysis of differential metabolites related to microbial load:①Differential metabolites significantly related to microbial load in positive ion mode were matched to choline,adenine,thymine,hypoxanthine and N-acetyl-D-mannose amine in this database.②Metabolites significantly related to microbial load in negative ion mode were matched to thymine,hypoxanthine,L-citrulline,deoxyuridine and choline phosphate in the database.(6)Metabolic pathway analysis of differential metabolites related to microbial load:①In positive ion model:The enrichment degree of purine metabolism was the highest,the impact factor of pyrimidine metabolism was the largest,and the enrichment degree of glucose metabolism was the lowest.The impact factor of amino acid metabolism was the smallest.②In negative ion mode:The degree of enrichment of pyrimidine metabolism is the highest,and its impact factor is the biggest.The enrichment degree of purine metabolism was the lowest and the impact factor was the smallest.Conclusion:This study showed that there was a correlation between some oral differential metabolites and microbial loads in HIV infected and non-HIV infected patients screened by LC-MS,suggesting that the difference of metabolites may be related to oral opportunistic infection.