The Metabolic Response Of Erythrocyte To Acute Exercise And The Adaptation Of Erythrocyte To Lactate Shuttle:Exploration Of Molecular Mechanisms Based On The NMR-based Technology

Sport-adaptation is the fundamental biological theory within sports trainings.By exploring the influence of sports training on motor ability and underlying mechanisms,we aim to diagnose,predict,and control the process and result of sports training,which makes sports training more operational and scientific.However,the theory of sports adaptation,especially in the biological mechanisms of human motor ability improvement aspect,still lacks experimental evidence and molecular mechanism related studies.Erythrocytes are the most numerous host cells in the human body and are visible components of the blood.As highly differentiated cells,it functions to transport life-essential oxygen and in exchange for carbon dioxide,which makes it an important research object in sports science fields.The metabolism of erythrocytes is unique,for instance,anaerobic glycolysis is the exclusive energy resource in erythrocytes.The effect of long-term exercise on the metabolism of erythrocytes may provide some shreds of evidence for the explanation of exercise adaptation theory.The main purpose of this article is to:(1)observing the acute response of erythrocytes during one-time submaximal intensity exercise by metabolomic methods;(2)investigating the erythrocytes'metabolic adaptation during long-term exercise training by analyzing the differences in lactate uptake and utilization between athletes and non-athletes populations in magnetic transfer(MT)experiment.Herein,we applied NMR based metabolomic analysis,magnetic transfer technology of isotope tracer,and molecular biological techniques across our research work.Chapter 1 introduces the metabolomic analysis results of the acute response of red blood cells to a one-time submaximal intensity exercise,in which the metabolic pattern changes,characteristic metabolites,and key metabolic pathways were identified.In brief,we found that a one-time submaximal intensity exercise disturbed the erythrocyte inherent metabolism and changed the metabolic pattern of red blood cells.Lactate and alanine are two main characteristic metabolites that increased significantly after exercise.These results were reciprocally confirmed in the plasma metabolomic analysis among these groups.In chapter 2,the erythrocytes metabolic adaptation induced by long-term exercise training was studied.In vivo experiments of erythrocytes were carried out by isolating erythrocytes from the athletes and non-athletes blood.We cultured erythrocytes with¹³C-lactate supplemented medium in which the lactate concentration simulates the blood lactate concentration under different exercise intensities,and compared the lactate influx of athletes and nonathletes populations.Meanwhile,with the help of NMR based living-cell isotope trace methods,we found the detailed downstream metabolic product of imported lactate.And the extracted membranes were subjected to western blot analysis,including MCT1,band3,and CD147.The result showed that under 8 m M lactate culture,the ratio of inside to outside lactate between athletes and nonathletes'erythrocytes was different,which was higher in athletes relative to nonathletes,and the expression of MCT1 and CD147 was consistently higher in athletes.Besides,we found that the imported lactate was further converted to pyruvate erythrocytes.In conclusion,the ¹H-NMR based metabolomics analysis result revealed that one-time submaximal intensity exercise changed the erythrocyte metabolic patterns,and long-term exercise could contribute to the lactate shuttling ability adaption in erythrocytes.This article provided new insights into the biological mechanisms involved in exercise adaptation researches and may advance our understanding of motion adaptation theory.