摘要
目的应用蛋白质组学技术,检测不同强度跑台运动后运动员尿液中差异蛋白的表达,探讨不同强度运动后尿液蛋白质组分的变化特点及其与人体免疫功能、运动性疲劳的关联性,为运动生化监控提供科学依据和实用方法。方法应用双向电泳法,分析8名男性运动员分别以55%、75%、85%、95%最大摄氧量(VO_(2max))的强度进行跑台运动后其尿液蛋白质组图谱的差异性表达,选取运动后差异蛋白表达量上调≥5倍且具有重复性的蛋白点,用基质辅助激光解吸电离串联飞行时间质谱法(MALDI-TOF-TOF-MS)进行质谱分析。结果经双向电泳确定的4种不同强度运动后尿液蛋白质组的差异性蛋白点共275个,其中,下调蛋白点85个,上调蛋白点190个;对差异蛋白表达量上调≥5倍且具有重复性的蛋白点进行质谱鉴定,共鉴定出29种蛋白,包括载脂蛋白、锌-α2-糖蛋白、免疫球蛋白、白蛋白、补体蛋白C3、甘露糖结合凝集素相关蛋白19、维生素D结合蛋白等;生物信息学分析结果表明,这些差异蛋白的功能主要与机体免疫调节和炎症反应的生物过程关系密切。结论蛋白质组学分析可以更好地诠释运动后尿液蛋白质组分的变化,其中差异表达的载脂蛋白、锌-α2-糖蛋白与运动后能量代谢有关,免疫球蛋白、白蛋白、补体蛋白C3、甘露糖结合凝集素相关蛋白19、维生素D结合蛋白与运动后免疫调节有关,这为考察运动训练后人体免疫功能和疲劳状态的变化提供了理论依据与应用方法。
Objectives Proteomics principles and techniques were used to detect and screen out the expression of differential proteins in urine of athletes after exercise with different intensities;the changes of urinary protein components and the correlations among the changes,human immune function and exercise fatigue were investigated,which provided scientific basis and practical methods for exercise biochemical monitoring.Methods Based on Two-dimensional electrophoresis(2-DE),differential expression of urine proteome of 8 male athletes was analyzed after exercises of 55%,75%,85% and 95% VO_(2max) intensity,respectively;the protein expressed levels which were up-regulated by at least 5-fold and with repeatability were screened and analyzed using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.Results A total of 275 differentially expressed protein points was detected after different exercise intensities,down-regulated protein points were 85 and up-regulated 190.After screening expressed levels which were up-regulated by at least 5-fold protein,a total of 29 protein was identified,showing a close relation to the biological processes of immune regulation and inflammatory response.Conclusions Proteomics better explains what happens to the urine proteome after different exercises,among which,apolipoprotein and zinc-α2-glycoprotein are related to lipid metabolism after exercise;immunoglobulin,albumin,complement component C3,mannose-binding lectin protein 19 and Vitamin D binding protein are related to immune regulation.The theoretical basis and application method are provided for the evaluation of the effects of exercise training on human immune function and fatigue.
作者
杨玲
蔺海旗
翁锡全
徐国琴
林文弢
YANG Ling;LIN Haiqi;WENG Xiquan;XU Guoqin;LIN Wentao(School of Physical Education,Shaoguan University,Shaoguan 512005,Guangdong,China;School of Physical Education,South China University of Technology,Guangzhou 510641,Guangdong,China;Laborotory of Exercise Biochemistry,Guangzhou Sport University,Guangzhou 510500,Guangdong,China)
出处
《上海体育学院学报》
CSSCI
北大核心
2021年第6期71-79,共9页
Journal of Shanghai University of Sport
基金
国家体育总局课题(2014B019)。
关键词
运动
尿液
蛋白质组
双向电泳
质谱
男运动员
exercise
urine
proteome
two-dimensional electrophoresis
mass spectrometry
male athlete
作者简介
第一作者:杨玲(ORCID:0000-0002-5485-0639),女,河北邢台人,华南师范大学博士研究生,韶关学院中级实验师,Tel.:18318491221,E-mail:475346214@qq.com;通信作者:林文弢(ORCID:0000-0002-0350-2957),男,广东惠来人,广州体育学院教授,博士生导师,Tel.:13070247816,E-mail:gtwtlin@163.com。
