Study On Hepatotoxicity Mechanism Of Anti-tuberculosis Drugs Based On Urine Metabolomics

Objective:through the study of the changes of urinary metabolics after the use of anti-tuberculosis drugs in tuberculosis patients,the possible mechanism of hepatotoxicity was explored in combination with the concentration of pyrazinamide and metabolites in urine.All of these studies provide a basis for clinical individualized medicationMethods:1?Approved by the Suzhou University Affiliated Infectious Diseases Hospital Ethics Committee to recruit clinically diagnosed tuberculosis patients.Patients were given standard anti-tuberculosis drugs including rifampicin,isoniazid,pyrazinamide and ethambutol.Collect the urine before and after the use of anti-tuberculosis drugs,and determine the relevant biochemical indicators.Of these patients,11 had liver damage and61 had no liver damage.In addition,17 urine samples of healthy subjects who met the age and sex matched those TB patients were collected.2?Based on UPLC-MS urine metabolomics assay platform,using ACQUITY UPLC? HSS T3 1.8 ?m(2.1 × 150 mm)column,0.1% formic acid water(A)-0.1%formic acid acetonitrile(B)The small molecular compounds in the urine were separated and the small molecular compounds in the urine were measured in a positive and negative ion mode using a Waters ACQUITY UPLC series Thermo LTQ Orbitrap XL while the MS/ MS fragments were analyzed for the compounds.The obtained data were subjected to peak identification,peak filtration and peak alignment to obtain information about the compound.The compounds were identified by the first-order precise molecular weight and secondary debris alignment in the database.The PCA and PLSDA models were used to determine the metabolic group differences between the groups.In order to explore the metabolic pathway of anti-tuberculosis drugs the difference in the compound was compared according to the VIP values in the PLS-DA model,the p-values of thecompounds between the groups,and the concentration ratio were determined by the first-order precise molecular weight and secondary debris alignment.Concentration ratio comparison Compounds differ to explore the effects of anti-tuberculosis drugs on metabolic pathways.3?The HPLC-UV method for the determination of pyrazinamide and its metabolites in urine was established.Ultimate? AQ-C18 column was used.The urine samples were eluted with mobile phase KH2PO4 aqueous solution and methanol gradient.The concentration of pyrazinamide and its metabolites in the urine was measured at the wavelength of 268 nm.In order to investigate the role of pyrazinamide in hepatotoxicity of anti-tuberculosis drugs,the relationship between the concentration of pyrazinamide and its metabolites and ALT and endogenous compounds was studied.Results:1 ? There were significant differences in metabolomics among healthy subjects,patients with newly diagnosed tuberculosis,patients without liver injury after treatment,and patients with liver injury after treatment.2?9 compounds were significantly higher than those of untreated patients,inclu-ding isonicotinic-acid,methyl fumaric acid(p <0.01),2-Hydroxypropylphosphonate,etc.18 compounds were significantly decr-eased after administration,respectively Succinylacetone,isocitrate,3-Hydroxy-2-me-thylpyridine-4,5-dicarboxylate,xanthine(p<0.01),7-Cyano-7-deazaguanine,etc.These mainly affect the body's arginine and proline metabolic pathways,purine metabolic pathway and tricarboxylic acid circulation pathway.3?The contents of 7 compounds with liver damaged group were significantly higher than those of non-liver injury group,including uric acid,cis-4-Octenedioic acid(p<0.01).The contents of 13 compounds decreased significantly in the liver injury group,including aconitic acid and hypoxanthine(p <0.01),etc.Metabolic pathway analysis showed that liver damage caused by anti-tuberculosis drugs was associated with pentose phosphate pathway,purine metabolic pathway and pyrimidine metabolic pathway.4?The metabolites of pyrazinamide in urine are associated with the product of the purine metabolic pathway.Metabolites of pyrazinamide such as 5-OH-PA,PA and5-OH-PZA were positively correlated with hypoxanthine in urine,the correlation coefficients were 0.33,0.36 and 0.49.5-OH-PZA and xanthine showed a moderate positive correlation,the correlation coefficient was 0.37.5-OH-PA,PA,5-OH-PZA in urine werenegatively correlated with uric acid,the correlation coefficients were-0.38,-0.45,-0.34.Conclusion:1 ? The hepatotoxicity caused by anti-tuberculosis drugs is related to the pentose phosphate pathway and the purine metabolic pathway.2?The participation of metabolites of pyrazinamide affects the pathway of purine metabolism,resulting in hepatotoxicity.