Research On The Metabolic Features Of Middle And Long Distance Runners In Heavy Load Training Stage And The Possible Mechanism Of Acupoint Stimulation Based On Metabolomics

Objectives:Study of middle-long-distance substance metabolism and energy metabolism has made a lot of good results,but most of the physiological and biochemical methods based on traditional preset some general macromolecular substances were studied,it is difficult to reflect comprehensively the sports training and competition of the metabolic integrity,systemic effects,therefore,urgent need to introduce a new concept and method,a more comprehensive and accurate reflection of middle-long-distance training material metabolism and energy metabolism in obtaining integrity and dynamics of change.Acupoint stimulation can stimulate the body's self-regulation function and has great potential in promoting the recovery of body function and improving the body's ability to exercise.Previous studies on acupoint stimulation often focus on the changes of a certain substance or several substances,and it is difficult to reflect the overall effect of acupoint stimulation on human body regulation.Metabolomics can analyze the obtained high-throughput biological information by"panoramic"scanning of metabolite changes,which is a powerful analytical method to reveal the influence of heavy load training on metabolism and acupoint stimulation regulation.In this study,the metabolomics method based on NMR was used to analyze and discuss the metabolic characteristics of middle and long-distance runners in the heavy load training phase,to find the key metabolites that affect the metabolic pathway changes of middle and long-distance runners in the heavy load training phase,and to construct the metabolomics atlas.To study the changes of metabolic pattern and molecular mechanism of long-term acupoint stimulation intervention after heavy load training for middle-distance runners,and to explain the possible mechanism of acupoint stimulation in the recovery of athletes'functional state from the perspective of metabolism.Methods:Eighteen male athletes of the long distance running team affiliated to Shanghai sports university were selected,all of whom were healthy.They were divided into the experimental group(LTA,9):acupoint stimulation group and the control group(LTR,9):non-acupoint stimulation group."Zusanli"(legs),"weizhong"(legs),"shenshu"and"guanyuan"points were selected for electroacupuncture stimulation of athletes in the experimental group(LTA)and the control group(LTR)(the control group was stimulated by 0.5-inch acupoint side opening)for 35 minutes every day for 4weeks.Three urine samples were collected on the morning of the training(Monday).Monday morning in the middle of training(two weeks later);Monday morning in late training(four weeks later).NoesyPr1d pulse[rd-90-t1-90-tm-90-acq]with presaturated PWP was used to detect the one-dimensional NOESY spectrum.All spectra under the condition of 25?with ultra-low temperature probe of Bruker(Karlsruhe,Germany)Avance III 600 MHz spectrometer are collected.Using MestReNova FID data processing software(version 12.0,Mestrelab Research lavrov).Fourier transform of FID with 0.3hz linewidth factor was used to improve the signal-to-noise ratio of the spectra,and then phase correction,baseline adjustment and spectrum peak alignment were performed on the spectra,and the methyl peak of the TSP was calibrated to 0.00 PPM.The peak height of each non-overlapping spectral peak was taken as the quantitative result of the spectral peak after the attribution of the metabolite,and then the data was normalized.The normalized data were normalized by UV scaling in simca-p~+14Principal component analysis(PCA),least squares-supervisory analysis(pls-da),orthogonal projection-supervisory analysis(opls-da)of potential structures are performed on software 14(Umetrics AB,Umea,Sweden).Results:(1)After heavy load training,the content of taurine,ascorbic acid,glucose,n-acetylglycoprotein,2-amino-hexamic acid and 2-hydroxyisobutyric acid in athletes'urine decreased significantly.The contents of glutamine,tyrosine,propylene glycol,lactic acid,dimethylglycine,valine,methylnicotinamide,alpha-ketoglutaric acid,alanine and formic acid increased significantly,mainly involving the changes of amino acid metabolism,energy metabolism,oxidative stress and the metabolic pathway of intestinal flora.(2)After acupoint stimulation,the content of n-acetylglycoprotein and phenylacetylglycine in athletes'urine increased;The contents of glutamine,citric acid,lactic acid,-ketoglutaric acid,tyrosine,3-aminoisobutyric acid,glycine and formic acid decreased.The main metabolic pathways of acupoint stimulation on athletes are amino acid metabolism,energy metabolism and intestinal flora metabolism.Conclusions:(1)There are significant differences in NMR profiles of urine samples at the beginning and end of heavy load training of middle-distance runners,which could screen out the key metabolites affecting the changes of metabolic pathway of middle-distance runners in heavy load training from the metabolomics analysis.(2)The metabolic characteristics of middle-distance runners in heavy load training stage are as follows:aerobic and oxidative metabolism plays the largest role;Glycolysis accounts for a large proportion of lactic acid accumulation;Amino acid metabolism was active and most amino acid catabolism was enhanced.Higher levels of oxidative stress.(3)Acupoint stimulation can play a good regulating role in energy metabolism,amino acid metabolism and intestinal flora metabolism of middle-and-long distance runners in heavy load training stage.(4)Acupoint stimulation is targeted,and the possible mechanism is that acupoint stimulation can enhance or inhibit the activity of enzymes in the corresponding metabolic pathway;The"bidirectional regulation"function of acupoint stimulation is to activate the body's inherent regulation function objectively.