基于LC-MS的精英赛艇运动员对高原训练早期反应的PO-053血浆代谢谱

Huan Gao, B. Gao, Zhijun Meng
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Train load is an important factor influence the stress reactions which isn’t like as altitude travel for normal people. What would happen in the body of athletes when altitude training? We still know very little. Metabonomic give us a new tool to understand the whole map of body’s reaction to altitude training. In this pilot study, we aimed to explore the plasma profiles changes of elite rowers in the early two-weeks training at 2300m plateau using metabolic tool based on LC-MS. \nMethods Sixteen male elite rowers(age 25.56±3.44y, height 189.06±5.37cm, weight 82.81±12.25kg, training years 10.31±2.52y) from China National team took part in this research. This altitude training camp was organized in the preparation period of a new season. It lasted six weeks. The acute phase (AP) which was the first three days after arriving at 2300m altitude base was composed by regenerative training and low-intensity aerobic training sessions. 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After a longer adaption, in the 14th day of altitude training, plasma benzamide, indole-3-acetaldehyde, phenethylamine, indole, 4-chlorbenzoic acid and benzoic acid still increased but the amplitude reduced compared with the 4th day. Ingenuity Pathways Analysis (IPA) suggested that the top 5 canonical pathways were tyrosine biosynthesis Ⅳ, phenylalanine degradationⅠ, protein kinase A signaling, tRNA charging and phenylalanine degradation Ⅳ in AP. In CP, the top 5 canonical pathways were tRNA charging, urate biosynthesis, tryptophan degradation Ⅹ, guanosine nucleotides degradation Ⅲ and adenosine nucleotides degradationⅡ. \nConclusions In this pilot study, we found 83 and 79 plasma feature in acute and chronic phase respectively. Considering the sharp elevation, plasma benzamide and indole-3-acetaldehyde which involved in the regulation of energy metabolism of brain may be the sensitive makers in acute adaption to altitude for athletes. 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引用次数: 0

摘要

目的到达高原后,由于空气干燥、强紫外线,特别是低氧等因素,对高原生物安全造成影响。生活在平原上的运动员在早期阶段往往表现出换气过度、心动过速、头晕、睡眠障碍、胃肠障碍、认知障碍等应激反应。除了这些应激反应外,一些研究报道碳水化合物和脂质代谢也发生了显著变化。急性缺氧可增强糖酵解,抑制肝糖异生。一些参与脂质氧化的基因表达下调。然而,研究也发现,在长期适应高原环境后,一些酶的活性增加,参与了肌肉中的有氧代谢。这些前后矛盾的结论使我们感到困惑。列车负荷是影响应激反应的重要因素,与正常人的高原旅行不同。高原训练时运动员的身体会发生什么变化?我们所知甚少。代谢组学为我们了解人体对高原训练反应的全貌提供了新的工具。在这项初步研究中,我们旨在利用基于LC-MS的代谢工具,探讨精英赛艇运动员在2300m高原训练前两周的血浆谱变化。方法选取16名中国国家赛艇运动员(年龄25.56±3.44y,身高189.06±5.37cm,体重82.81±12.25kg,训练年龄10.31±2.52y)为研究对象。这次高原训练营是在新赛季的准备阶段组织的。它持续了六个星期。急性期(AP)是到达2300m海拔基地后的前三天,由再生训练和低强度有氧训练组成。在接下来的10天(慢性期,CP)中,包括三组低重量阻力训练在内的大多数训练的强度都是低到中等有氧。静息半天后,取静脉血5 ml入肝素抗凝管,4℃8000rpm离心15分钟,于第4天、第14天上午分离血浆。血浆保存于-80℃,用LC-MS测定代谢谱。使用Compound Discoverer 2.0软件(Thermo)对数据进行特征提取和预处理,然后使用excel 2010软件将数据归一化并编辑成二维数据矩阵,包括Retention time(RT)、Compound Molecular Weight (compMW)、Observations(samples)和峰强度。编辑后的特征数据使用SIMCA-P软件(Umetrics AB, Umea,瑞典)进行多变量分析(MVA)。结果与高原前相比,AP和CP在(ESI+)离子模式下分别发现26和30个特征,在(ESI-)离子模式下分别发现57和49个特征。AP与CP比较,(ESI+)离子模式下有46个特征,(ESI-)离子模式下有67个特征。AP患者血浆苯甲酰胺和吲哚-3-乙醛分别升高35.16倍和16.54倍。血浆中苯乙胺、苯酚、吲哚、哌啶、亮氨酸、4-氯苯甲酸和苯甲酸较高原前增加4.55 ~ 8.22倍。AP降低幅度最大的前3位为癸二酸二丁酯、亚苯甲氧基次黄嘌呤和硫酸氢胆固醇,分别降低1.76 ~ 3.85倍。适应时间较长后,高原训练第14天,血浆苯甲酰胺、吲哚-3-乙醛、苯乙胺、吲哚、4-氯苯甲酸和苯甲酸仍升高,但幅度较第4天有所降低。创新途径分析(Ingenuity Pathways Analysis, IPA)表明,AP中前5个典型途径是酪氨酸生物合成Ⅳ、苯丙氨酸降解Ⅰ、蛋白激酶A信号传导、tRNA充电和苯丙氨酸降解Ⅳ。CP中前5个典型途径是tRNA充电、尿酸生物合成、色氨酸降解Ⅹ、鸟苷核苷酸降解Ⅲ和腺苷核苷酸降解Ⅱ。结论本研究在急性期和慢性期分别发现83例和79例血浆特征。血浆苯甲酰胺和吲哚-3-乙醛参与大脑能量代谢的调节,可能是运动员急性高原适应的敏感因素。随着时间的延长,增加幅度在第14天开始下降。提示大脑能量代谢可能发生显著变化。在高原训练中,尤其在高原训练初期,应重视中枢神经系统的训练。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
PO-053 Plasma Metabolic Profiles of Elite Rowers Response to the Early Phase of Altitude Training Based on LC-MS
Objective When arriving at altitude, because of the dry air, strong ultraviolet and especially the low oxygen et.al., athletes who living on the plain always exhibited kinds of stress reactions such as hyperventilation, tachycardia, dizziness, sleep-disorder, gastrointestinal disturbance and cognitive impairment in the early phase. Except these stress reactions, some studies reported the carbohydrate and lipid metabolism show significant change. Acute exposure to hypoxia could strengthen the glycolysis and suppress hepatic gluconeogenesis. The expression of some genes involved in lipid oxidation was down-regulated. However, it’s also found the increased activity of some enzymes took part in aerobic metabolism in muscle after long term acclimatization to altitude. These inconsistent conclusions make us confusion. Train load is an important factor influence the stress reactions which isn’t like as altitude travel for normal people. What would happen in the body of athletes when altitude training? We still know very little. Metabonomic give us a new tool to understand the whole map of body’s reaction to altitude training. In this pilot study, we aimed to explore the plasma profiles changes of elite rowers in the early two-weeks training at 2300m plateau using metabolic tool based on LC-MS. Methods Sixteen male elite rowers(age 25.56±3.44y, height 189.06±5.37cm, weight 82.81±12.25kg, training years 10.31±2.52y) from China National team took part in this research. This altitude training camp was organized in the preparation period of a new season. It lasted six weeks. The acute phase (AP) which was the first three days after arriving at 2300m altitude base was composed by regenerative training and low-intensity aerobic training sessions. During the next ten days (chronic phase, CP), the intensity of most sessions including three low-weight resistance training sets were low to middle aerobic. After resting half day, 5 ml venous blood was collected into heparin anticoagulant tubes and then centrifuged at 8000rpm in 4℃ lasting 15 minutes to separate plasma in the 4th day and 14th morning. Plasma was stored in -80℃ to measure metabolic profile by LC-MS. The data was performed feature extraction and preprocessed with Compound Discoverer 2.0 software (Thermo), and then normalized and edited into two-dimensional data matrix by excel 2010 software, including Retention time(RT), Compound Molecular Weight (compMW), Observations(samples) and peak intensity. The feature data after editing were performed Multivariate Analysis (MVA) using SIMCA-P software (Umetrics AB, Umea, Sweden). Results Compared with pre-altitude, 26 and 30 features at (ESI+) ion mode, 57 and 49 features at (ESI-) ion mode were found in the AP and CP respectively. Compared AP with CP, there were 46 features at (ESI+) ion mode and 67 features at (ESI-) ion mode. In AP, plasma benzamide and indole-3-acetaldehyde increased 35.16 fold and 16.54 fold respectively. Plasma phenethylamine, phenol, indole, piperidine, leucine, 4-chlorbenzoic acid and benzoic acid increased 4.55 to 8.22 fold compared with pre-altitude. Top three decreased features were dibutyl sebacate, arbinosylhypoxanthine and cholesterol hydrogen sulfate which decreased 1.76 to 3.85 fold in AP. After a longer adaption, in the 14th day of altitude training, plasma benzamide, indole-3-acetaldehyde, phenethylamine, indole, 4-chlorbenzoic acid and benzoic acid still increased but the amplitude reduced compared with the 4th day. Ingenuity Pathways Analysis (IPA) suggested that the top 5 canonical pathways were tyrosine biosynthesis Ⅳ, phenylalanine degradationⅠ, protein kinase A signaling, tRNA charging and phenylalanine degradation Ⅳ in AP. In CP, the top 5 canonical pathways were tRNA charging, urate biosynthesis, tryptophan degradation Ⅹ, guanosine nucleotides degradation Ⅲ and adenosine nucleotides degradationⅡ. Conclusions In this pilot study, we found 83 and 79 plasma feature in acute and chronic phase respectively. Considering the sharp elevation, plasma benzamide and indole-3-acetaldehyde which involved in the regulation of energy metabolism of brain may be the sensitive makers in acute adaption to altitude for athletes. As the extension of time, the increased amplitude came down in the 14th day. It suggested that the energy metabolism of brain may take significant change. Central nerve system should be paid more attention during altitude training especially in the early phase.
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