Systematic Evaluations Of Vancomycin-Induced Toxicity In Rats Based On Metabolomics

Objective: Using gas chromatography-mass spectrometry（GC-MS）to detect healthy rats and rats injected with vancomycin intraperitoneally,fi nd out the significantly different metabolites between the two groups,co nduct systematic evaluation,and explore the mechanism of vancomycin t oxicity to various tissues,so as to provide new ideas for clinical medicat ion.Materials and Methods:（1）6-week-old male KM mice were random ly divided into experimental group and control group（n=9）.Continuous i ntraperitoneal injection of vancomycin（400mg/kg/day）for 7 days.（2）After the metabolites were separated by GC-MS method,the orig inal data was obtained.Agilent Mass Hunter Quantitative software and Un knowns Analysis software was used to identify the compound correspond ing to the detected peak.Compare the database with NIST14.0 mass spe ctrometry library to extract known metabolites for subsequent biological i nformation analysis.Principal component analysis（PCA）and orthogonal partial least squares discriminant analysis（OPLS-DA）were used to analy ze the processed data to find the difference between the control group a nd the experimental group,200 response ranking was used to evaluate th e stability of OPLS-DA models.To comparemetabolites between two gro ups,univariate statistical analysis was carried out using Two-tailed Stude nt’s t-tests with SPSS 19.0 software（SPSS Inc,Chicago,IL,USA）.Comp ounds with variable importance in projection（VIP）values > 1.0 and two-tailed student’s t-test P values < 0.05 were deemed potential discriminan t metabolites.Metabo Analyst 4.0 and KEGG were used to analyze pathw ays.Pathways with raw P values < 0.05 and impact values > 0 and Ra w P<0.5,impact >0 were defined as significant.Results: Compared with the control group,we found different metab olites in the heart,liver,kidney,spleen,brain,hippocampus,inner ear,lun g,intestine,and serum in the vancomycin group.The number of differen t metabolites in different tissues was 23 in heart（mainly amino acid,eth anolamine,uracil,malic acid,pyroglutamic acid,3-hydroxypyridinoic acid,o-ethanolamine phosphate,arachidonic acid,glycerol,4-hydroxyproline,o leic acid and docosahexaenoic acid）,16 in liver（mainly amino acid,D-ri bose,uracil and urea）,27 in kidney（mainly amino acids,uracil,nicotin amide,ribol,o-ethanolamine phosphate,tin inositol and inosine）,18 in s pleen（mainly amino acids γ-Aminobutyric acid,myristic acid,inositol,3-methyl-2-oxovanoic acid,Mg（0:0/18:0/0:0）,formamide,pyroglutamic ac id,stearic acid,arachidonic acid,Mg（16:0/0:0/0:0）,bile solid）,14 in bra in（mainly amino acids,citric acid,docosahexaenoic acid and oleic acid）,9 in hippocampus（mainly creatinine,2-aminobenzoic acid,docosahexae noic acid,Mg（16:0/0:0）,n-dodecane）,7 in inner ear（mainly undecane,4-hydroxybutyric acid,urea,inositol,arachidonic acid and PCBs）,9 in lu ng（mainly D-mannose,uracil,urea,citrulline,succinic acid,myristic acidand isooleic acid）,11 in intestine（mainly L-lactic acid,n-decane,nonan e,tetradecane,tridecanol,myristic acid,tin inositol,palmitic acid,stearicacid,Mg（0:0/18:0/0:0）Sulfite）and 13 in serum（mainly amino acids,i nositol and urea）.Analyzed them separately,15 metabolic pathways were found,including Arginine Biosynthesis,Butanoate metabolism,Glutathione metabolism,Glycerophospholipid metabolism,Glycine serine and threonin e metabolism,Glyoxylate and dicarboxylate metabolism,D-Glutamine and D-glutamate metabolism,Phenylalanine-tyrosine and tryptophan biosynthesis,Alanine aspartate and glutamate metabolism,Phenylalanine metabolism,Glycerolipid metabolism,Arginine and proline metabolism,Fatty Acid Bi osynthesis,Sulfur Metabolism Arginine Biosynthesis.Conclusion:（1）Vancomycin can affect many tissues and organs of human body.（2）After vancomycin treatment,the differential metabolites were mainly concentrated in liver,kidney,spleen,brain and heart.16,24,18,14 and 23 differential metabolites were found respectively.（3）The changes of metabolic pathways in the experimental group mainly involve d 15 metabolic pathways.（4）The injury caused by vancomycin exposuremay be realized through oxidative stress injury,neurotoxicity and the ac cumulation of amino acids and fatty acids in multiple organs.