GC-MS Based Metabonomics Study Of Serum And Urine From Weightlifters And Middle-distance Runners

ObjectiveThe purpose of this study is to establish a gas chromatography-mass spectrometry (GC-MS) based metabonomics analytical method of serum and urine to study metabonomic information from weightlifters and middle-distance runners. The metabonome differences in serum and urine between two events and among before, medium-term and after stage training, the before and after different load of single training course for every event are compared to verify the feasibility of metabonomic technology applied in sports medicine field and to explore the characteristics of global metabolic responses induced by sports training . Then we hope to provide a new idea and method for sports medicine research from metabolites level.MethodsEight male weightlifters of JiangSu province weightlifter team and eight male middle-distance runners of the JiangSu province track and field team were enrolled in a 6 weeks stage training and a crossover study for different load of single training course. Seven blood and urine samples were drawn for each subject: before, medium-term and after of stage training, befor and after of heavy and low load training. Biofluid samples were analyzed by GC-MS based metabonomic technology combined with data analysis technology such as denosing, peak sorting, chromatographic peak area calibration, multivariable statistical analysis(principal component analysis,PCA and partial-least squares-discriminant analysis,PLS-DA) and metabolites identification use of NIST 2005. The RBC, HGB, BUN, CK, C and T were also analyzed by conventional biochemistry technology.Results(1) GC-MS based metabonomic analytical method was established and which was obtained by optimizing both separation condition of gas chromatography and mass spectrometer parameters, and of great repeatability, precision, days stability and freeze-thaw stability.(2) The serum and urine metabonomic data of weightlifters and middle-distance runners from before, medium-term and after of stage training shown that: there are fourteen metabolites in serum and sixteen metabolites in urine have great different concentration between weightlifters and middle-distance runners. And there are twenty metabolites (D-ribose ,α-D-glucopyranose ,β-D-glucopyranose , and so on) in serum and twenty metabolites(L-histidine, arabitol, butyric acid, and so on) in urine have significant change for weightlifters, while nine metabolites(glycine, phosphate, D-myo-inositol, and so on) in serum and six metabolites(creatinine, L-histidine, L-tyrosine, and so on) in urine have significant change for middle-distance runners compared between before training and medium-term training, medium-term training and after training and before training and after training.(3) The serum and urine metabonomic data of weightlifters and middle-distance runners from before and after of different load single training course shown that: phosphate,α-tocopherol, 9,12-octadecadienoic acid and acetic acid in serum and creatinine,α-D-glucopyranose, D-glucopyranose, sedoheptulose andβ-D-glucopyranose in urine have significant change after heavy load single training for weightlifters, while urea, galactopyranose, propanoic acid, myo-inositol,α-tocopherol, stearic acid, 5,8,11,14,17-eicosapentaenoic acid and myristic acid in serum and butanedioic acid and hexadecanoic acid in urine have significant change after heavy load single training for middle-distance runners. After low load single training, for weightlifters, the concentration of 9,12-octadecadienoic acid and myo-inositol in serum and arabinitol, sedoheptulose and glucopyranose in urine decreased; for middle-distance runners, the concentration of phosphate, acetic acid, stearic acid in serum decreased but N-methyl nicotinimidate increasd, and the concentration of hexadecanoic acid in urine increased.Conclusions(1) The PLS-DA analytical modle is better than PCA's for metabonomic data process.(2) Carbohydrate, amino acid, free fatty acid and other metabolites related to glycogenolysis, gluconeogenesis, TCA cycle, urea cycle and ketone body synthesis are change induce by exercise, and the more time continued and load to take, the more change was induced.(3) Different events athletes (weightlifters and middle-distance runners) have different metabonomic profiling of serum and urine.(4) Serum combined with urine matabonomics study can provide more comprehensive information for sports medicine research.(5) The technology of metabonomics is greater sensitive and appropriate than the conventional biochemical method. Joint each other can provide more comprehensive and reliable information for sports training monitoring.