格斗对抗性项目代谢调节与适应的核磁共振代谢组学研究
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摘要
运动训练和竞赛对新陈代谢产生系统性、整体性的影响。通过几个指标的检测难以反映运动产生的代谢变化的全貌,因此,引入系统生物学的思路和方法来研究运动训练的代谢问题,具有明显的方法学优势。代谢组学(metabonomics/metabolomics)是继基因组学和蛋白质组学之后发展起来的一门新学科,通过“全景式”地扫描代谢物的变化,对所获得的高通量生物学信息进行分析,以研究运动对机体的整体影响,是揭示运动过程中代谢变化规律的强有力的分析方法。
本文采用基于核磁共振(NMR)的代谢组学、模式识别和化学计量学等方法,选择了击剑和散打等两个格斗对抗性项目(以下简称格斗项目),研究：1)竞技比赛中代谢的调节及其机制；2)长期训练和比赛引起的代谢适应及其机制。
绪论主要对研究问题进行综述,指出目前研究存在的不足并进行展望。同时对本文研究思路进行了概括性的论述。
第一章第一节研究了击剑项目竞赛中的代谢调控及分子机制。观察了全国击剑赛花剑运动员比赛前后尿液中代谢物的变化。结果显示,比赛前后运动员的代谢模式发生了明显的变化,造成这一变化的特征性代谢物涉及肌酸代谢、三羧酸循环、糖代谢、氨基酸代谢、酮体代谢、嘌呤核苷酸代谢等途径,N-氧化三甲胺(TMAO)浓度下降,氧化应激增强。此外,一些氨基酸类神经递质的含量发生了明显的变化。研究提示,花剑比赛是以有氧供能为主的运动项目；神经系统的活动、体内氨的生成和清除对于花剑竞赛具有重要的意义。本研究为花剑竞赛和训练中的监控提供了新的技术手段。
第一章第二节研究散打竞赛的代谢调控及分子机制。同时,我们也研究了训练中常用的三种类型的训练(一般性体能训练、专项体能训练和队内教学比赛)对代谢模式的影响,并与正式比赛后代谢模式的变化进行比较,分析不同类型训练方案的有效性。结果表明,正式比赛中代谢模式的变化大于三种类型的训练,主要体现在整体能量代谢和内源性神经递质的变化更明显,提示神经系统活动加强会造成能量代谢的增加,这是格斗项目重要的代谢特征,也是格斗项目训练方案设计时需要引起高度关注的问题。此外,NMR代谢组学技术和方法可以为训练方案的选择、制定和效果验证提供全新的思路。
第二章研究了格斗运动员(散打和击剑)长期训练所致的代谢适应的特征及机制。分别比较格斗运动员与普通人之间、以及格斗运动员与短跑运动员之间尿液中代谢物的差异,连续3周收集周一上午的晨尿,寻找在连续3次的检测中均发现组间差异的特征性代谢物,分析长期训练引起的相对稳定的代谢特征,并讨论其形成机制。结果显示,格斗运动员在肌酸代谢、氨基酸代谢和肠道菌群代谢方面具有不同于普通人的一般性适应特征,而在能量储备种类、占优势的代谢类型、抗氧化能力、神经递质的浓度等方面具有不同于短跑运动员的专项性适应特征。研究提示,代谢适应是长期专项训练和竞赛累积的效应,是运动员适应专项需要的表现。本研究筛选出的代谢物可以作为格斗对抗性项目训练监控、优秀运动员选材、训练效果评价的潜在的生化指标。
第三章跟踪观察了一例由于赛前快速减体重造成运动能力下降的典型个案。第一节和第二节分别动态观察了这名散打运动员在减体重过程中的不同阶段,静态(安静状态下)和动态(训练前后及恢复前后)的代谢模式的变化及其时间轨迹。结果显示,在快速减体重过程中,体内整体的能量代谢经历了先升后降的过程；减体重后期酮体代谢占据重要地位。禁食状态下进行训练,机体不仅出现代谢紊乱,而且内环境稳态受到破坏,并危害身体健康。基于NMR的代谢组学技术,比传统的监控指标分析能够更加敏感和全面地进行个体训练监控和评价,为个体化训练监控和科技服务提供了强有力的技术平台和研究方法。
Sports training and competition have a systemic impact on metabolism, which can not be clari Sports training and competition have a systemic impact on metabolism. Metabolism of sport and training has been intensively studied nearly100years through traditional biochemical methods. However, the systemic effects of training cannot be clarified by several indicators. Metabonomics (Metabolomics) is proven to be a powerful strategy to reveal the mechanisms involved in systemic metabolic responses induced by sports training and competition through studying global changes of endogenous metabolites.
In this thesis, nuclear magnetic resonance (NMR)-based metabonomics, along with pattern recognition was employed to reveal:1) metabolic regulation during competitions of combat sport;2) metabolic adaptation induced by long-term training in fencers and Sanda players.
In the introduction, I reviewed the recent literature on using a metabonomic approach in sport and exercise field, analyzed the major findings, shortcomings, and its potential applications in the future.
In the first section of Chapter I, we studied the metabolic regulation during a national fencing competition, analyzed its molecular mechanisms. Urinary metabolites were examined by NMR before and after the competition. Metabolic profiling revealed pronounced shifts in the several metabolic pathways during the competition, including the citric acid cycle, metabolism of creatine, amino acid, ketone body and purine nucleotide. Trimethylamine-N-oxide (TMAO) concentration decreased, and metabolic biomarkers of oxidative stress increased after the competition. Interestingly, we found several amino acid neurotransmitters which suggested an inhibition of nervous regulation. Furthermore, the generation and removal of ammonia may be important in foil competition.
In the second section of Chapter I, we studied the metabolic regulation during a Sanda competition. Furthermore, we examined metabolic reactions of three training procedures, and compared them with those of a Sanda competition, so as to analyze the effectiveness of the training procedures. Results showed that the metabolic patterns were different between them. In the Sanda competition, the global energy metabolism changed much more than training. Amino acid contributed more in the competition than in the training procedures. Heart rate variability analysis also expressed a lower down-regulation of parasympathetic activity in the competition, indicating the energy consumption increase derived from the neural activity might be an important feature in combat sport.
Chapter Ⅱ focused on the metabolic adaptation by long-term specific combat training and its underlying regulatory mechanisms. Fencing and Sanda's athletes were compared to a gender, age-paired healthy control, and to sprinters as athlete control. Fasting urinary samples were collected in three consecutive Monday morning, to find metabolites, which were distinctive between the groups. We tried to find biomarkers of metabolic adaptation in combat sport. Results showed that the metabolic patterns of the combat athletes were different from healthy control, and from sprinting athletes as well. Some metabolic biomarkers were identified which relevant to creatine metabolism, amino acid metabolism and intestinal flora metabolism between combat athletes with non-athletes. Type of energy storage, the dominant metabolic pathways, antioxidant capacity, neurotransmitter concentrations were different between combat sport athletes with sprinting athletes. The metabolic adaptations are accumulating effects of long-term discipline-specific training and competition. The identified metabolites can serve as potential biochemical indicators in training monitoring and evaluating.
Chapter Ⅲ is a case report. A Sanda player who experienced a rapid weight loss which led to a significant decline of sport performance before a competition. Section1and2described respectively the metabolism pattern changes in the steady-state and in the dynamic state respectively during the rapid weight loss procedure. Results showed that energy metabolism of steady-state up-regulated in the early stage, and then down-regulated in the late stage. The enhanced oxidation of ketone bodies for energy consumption was important primarily in the late stage. During the period of very low energy intake, sports training resulted in a metabolic disorder and disruption of the stability of internal environment, which might do harm to health.
本文采用基于核磁共振(NMR)的代谢组学、模式识别和化学计量学等方法,选择了击剑和散打等两个格斗对抗性项目(以下简称格斗项目),研究：1)竞技比赛中代谢的调节及其机制；2)长期训练和比赛引起的代谢适应及其机制。
绪论主要对研究问题进行综述,指出目前研究存在的不足并进行展望。同时对本文研究思路进行了概括性的论述。
第一章第一节研究了击剑项目竞赛中的代谢调控及分子机制。观察了全国击剑赛花剑运动员比赛前后尿液中代谢物的变化。结果显示,比赛前后运动员的代谢模式发生了明显的变化,造成这一变化的特征性代谢物涉及肌酸代谢、三羧酸循环、糖代谢、氨基酸代谢、酮体代谢、嘌呤核苷酸代谢等途径,N-氧化三甲胺(TMAO)浓度下降,氧化应激增强。此外,一些氨基酸类神经递质的含量发生了明显的变化。研究提示,花剑比赛是以有氧供能为主的运动项目；神经系统的活动、体内氨的生成和清除对于花剑竞赛具有重要的意义。本研究为花剑竞赛和训练中的监控提供了新的技术手段。
第一章第二节研究散打竞赛的代谢调控及分子机制。同时,我们也研究了训练中常用的三种类型的训练(一般性体能训练、专项体能训练和队内教学比赛)对代谢模式的影响,并与正式比赛后代谢模式的变化进行比较,分析不同类型训练方案的有效性。结果表明,正式比赛中代谢模式的变化大于三种类型的训练,主要体现在整体能量代谢和内源性神经递质的变化更明显,提示神经系统活动加强会造成能量代谢的增加,这是格斗项目重要的代谢特征,也是格斗项目训练方案设计时需要引起高度关注的问题。此外,NMR代谢组学技术和方法可以为训练方案的选择、制定和效果验证提供全新的思路。
第二章研究了格斗运动员(散打和击剑)长期训练所致的代谢适应的特征及机制。分别比较格斗运动员与普通人之间、以及格斗运动员与短跑运动员之间尿液中代谢物的差异,连续3周收集周一上午的晨尿,寻找在连续3次的检测中均发现组间差异的特征性代谢物,分析长期训练引起的相对稳定的代谢特征,并讨论其形成机制。结果显示,格斗运动员在肌酸代谢、氨基酸代谢和肠道菌群代谢方面具有不同于普通人的一般性适应特征,而在能量储备种类、占优势的代谢类型、抗氧化能力、神经递质的浓度等方面具有不同于短跑运动员的专项性适应特征。研究提示,代谢适应是长期专项训练和竞赛累积的效应,是运动员适应专项需要的表现。本研究筛选出的代谢物可以作为格斗对抗性项目训练监控、优秀运动员选材、训练效果评价的潜在的生化指标。
第三章跟踪观察了一例由于赛前快速减体重造成运动能力下降的典型个案。第一节和第二节分别动态观察了这名散打运动员在减体重过程中的不同阶段,静态(安静状态下)和动态(训练前后及恢复前后)的代谢模式的变化及其时间轨迹。结果显示,在快速减体重过程中,体内整体的能量代谢经历了先升后降的过程；减体重后期酮体代谢占据重要地位。禁食状态下进行训练,机体不仅出现代谢紊乱,而且内环境稳态受到破坏,并危害身体健康。基于NMR的代谢组学技术,比传统的监控指标分析能够更加敏感和全面地进行个体训练监控和评价,为个体化训练监控和科技服务提供了强有力的技术平台和研究方法。
Sports training and competition have a systemic impact on metabolism, which can not be clari Sports training and competition have a systemic impact on metabolism. Metabolism of sport and training has been intensively studied nearly100years through traditional biochemical methods. However, the systemic effects of training cannot be clarified by several indicators. Metabonomics (Metabolomics) is proven to be a powerful strategy to reveal the mechanisms involved in systemic metabolic responses induced by sports training and competition through studying global changes of endogenous metabolites.
In this thesis, nuclear magnetic resonance (NMR)-based metabonomics, along with pattern recognition was employed to reveal:1) metabolic regulation during competitions of combat sport;2) metabolic adaptation induced by long-term training in fencers and Sanda players.
In the introduction, I reviewed the recent literature on using a metabonomic approach in sport and exercise field, analyzed the major findings, shortcomings, and its potential applications in the future.
In the first section of Chapter I, we studied the metabolic regulation during a national fencing competition, analyzed its molecular mechanisms. Urinary metabolites were examined by NMR before and after the competition. Metabolic profiling revealed pronounced shifts in the several metabolic pathways during the competition, including the citric acid cycle, metabolism of creatine, amino acid, ketone body and purine nucleotide. Trimethylamine-N-oxide (TMAO) concentration decreased, and metabolic biomarkers of oxidative stress increased after the competition. Interestingly, we found several amino acid neurotransmitters which suggested an inhibition of nervous regulation. Furthermore, the generation and removal of ammonia may be important in foil competition.
In the second section of Chapter I, we studied the metabolic regulation during a Sanda competition. Furthermore, we examined metabolic reactions of three training procedures, and compared them with those of a Sanda competition, so as to analyze the effectiveness of the training procedures. Results showed that the metabolic patterns were different between them. In the Sanda competition, the global energy metabolism changed much more than training. Amino acid contributed more in the competition than in the training procedures. Heart rate variability analysis also expressed a lower down-regulation of parasympathetic activity in the competition, indicating the energy consumption increase derived from the neural activity might be an important feature in combat sport.
Chapter Ⅱ focused on the metabolic adaptation by long-term specific combat training and its underlying regulatory mechanisms. Fencing and Sanda's athletes were compared to a gender, age-paired healthy control, and to sprinters as athlete control. Fasting urinary samples were collected in three consecutive Monday morning, to find metabolites, which were distinctive between the groups. We tried to find biomarkers of metabolic adaptation in combat sport. Results showed that the metabolic patterns of the combat athletes were different from healthy control, and from sprinting athletes as well. Some metabolic biomarkers were identified which relevant to creatine metabolism, amino acid metabolism and intestinal flora metabolism between combat athletes with non-athletes. Type of energy storage, the dominant metabolic pathways, antioxidant capacity, neurotransmitter concentrations were different between combat sport athletes with sprinting athletes. The metabolic adaptations are accumulating effects of long-term discipline-specific training and competition. The identified metabolites can serve as potential biochemical indicators in training monitoring and evaluating.
Chapter Ⅲ is a case report. A Sanda player who experienced a rapid weight loss which led to a significant decline of sport performance before a competition. Section1and2described respectively the metabolism pattern changes in the steady-state and in the dynamic state respectively during the rapid weight loss procedure. Results showed that energy metabolism of steady-state up-regulated in the early stage, and then down-regulated in the late stage. The enhanced oxidation of ketone bodies for energy consumption was important primarily in the late stage. During the period of very low energy intake, sports training resulted in a metabolic disorder and disruption of the stability of internal environment, which might do harm to health.
引文
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