Analysis Of The Influence Of HAART On The Oral Metabolites Of HIV Infected Patients

Objective:Highly active antiretroviral therapy(HAART)has greatly improved the life expectancy of HIV infected patients.However,long-term use of HAART is associated with metabolic abnormalities and increases the risk of kidney,liver and cardiovascular dysfunction.Metabolomic analysis of the changes of metabolites in patients subsequent to their usage of antiviral drugs helps to identify biomarkers associated with drug use.At present,there are few studies about the changes of metabolites in oral fluid of HIV patients after using HAART.This project collected the oral fluid of HIV infected patients in Yunnan before HAART usage and 6 months,12 months and 18 months after the use of HAART,and analyzed the metabolites in these patients.The purpose of this project was to explore the effect of HAART on the oral metabolites of the patients.Methods:A total of 30 HIV-infected persons were included in the group.The oral fluid collected before HAART usage was labeled as Group A,oral fluid specimens of the follow-up patients were collected sequentially at 6,12,and 18 months after drug administration and marked as Groups,D,E and F,respectively.18 months subsequent to HAART usage,10 patients no longer participated in our follow-up.For the 20 patient samples which we followed at all time points,LC-Q-TOF-MS(combining liquid chromatography-mass spectrometry(LC-MS)with quadrupole flight time(Q-TOF)platform positive ion mode(POS)and negative ion mode(NEG)were used to detect metabolites to conduct preprocessing,univariate statistical analysis,multivariate statistical analysis(including principal component analysis(PCA)and orthogonal partial least squares discriminant analysis(OPLS-DA),etc.on the obtained data.Finally,a meaningful inter-group differential metabolite was screened and the metabolic pathways affected by HAART were analyzed.Results:In the comparison among Group D(6 months after HAART usage),Group E(12 months after HAART usage),Group F(18 months after HAART usage)and Group A(pretreatment group),first,the main components of all the samples were similar by PCA analysis,and the OPLS-DA analysis showed that the metabolic difference between the samples in groups D,E and F and those in group A was significant.Meanwhile,the PCA models of Groups D,E and F showed no significant difference among the three groups.The differential metabolites were screened out by VIP greater than 1 and P value less than 0.05,fold change>2 or<0.5.Comparison between Groups D,E,F and Group A showed in the oral fluids of the three groups which received HAART,8 metabolites,including N-acetyl-L-omithine,inosine,5-deoxyadenosine,5-methylthioadenosine,2-hydroxypyridine,uracil,alanine threonine peptide,and tryptophyl-tyrosine,were down-regulated.Another 8 differential metabolites,including Isoleucine aspartate dipeptide,N-(omega)-Hydroxy arginine,L-Ny-monomethyl arginin,2-Hydroxyadenine,prolyl arginine,quinone Acetyl histidine,arginine isoleucine small peptide,and amino acid proline,were upregulated.Finally,the differential metabolites KEGG and differential metabolite pathway analysis focused on amino acid metabolism,glucose metabolism,nucleotide metabolism(pyrimidine metabolism,purine metabolism),pyruvate metabolism(aform of sugar,fat and protein conversion process)and fat metabolism(fatty acid metabolism,glycerophospholipid metabolism).The changes in the three metabolic pathways of Group D,E and F samples were mainly sugar metabolism,amino acid metabolism and sphingolipid metabolism.Conclusion:In this study,LC-Q-TOF-MS was used to detect oral fluid metabolites before and after use of HAART in HIV-infected individuals.The results show that there are many levels of metabolites change.The results of the study has significant implications for further understanding of the impact of HAART on the metabolism of HIV-infected host and the possible mechanism of such impact.