目的 阐明黄芪复合配方的抗疲劳作用及代谢组学机制.方法 雄性昆明小鼠随机分为10组,分别为空白组、黄芪提取物低、中、高剂量组、抗疲劳配方低、中、高剂量组、黄芪复合配方低、中、高剂量组,连续灌胃给样29天后,每组进行负重游泳实验.连续灌胃给样30天,小鼠力竭游泳后取材,分离血清检测尿素氮、乳酸、乳酸脱氢酶含量;分离肌肉检测肌糖原、Na+-K+-ATP酶和Ca2+-Mg2+-ATP酶含量;随机选取空白组和黄芪复合配方中剂量组各6只小鼠的骨骼肌,采用超高效液相色谱——四极杆飞行时间质谱联用的代谢组学技术(ultra-high performance liquid chromatography coupled to quadruple time-of-flight mass spectrometry,UHPLC-QTOFMS)、主成分分析和偏最小二乘法—判别分析的多元统计分析方法,筛选出黄芪复合配方抗疲劳的内源性差异代谢物,并确定与差异代谢物相关的代谢通路,探究该黄芪复合配方对缓解运动性疲劳的作用机制.结果 各组小鼠给样后,负重游泳实验结果显示,与空白组相比,黄芪提取物各剂量组、抗疲劳配方各剂量组、黄芪复合配方各剂量组小鼠力竭游泳时间均明显延长(P＜0.05,P＜0.001);血清及肌肉指标检测显示,与空白组相比,抗疲劳配方高剂量组、黄芪复合配方低、中、高剂量组肌糖原含量显著升高(P＜0.05,P＜0.01);除黄芪提取物组外各组血乳酸均显著降低(P＜0.05,P＜0.01,P＜0.001);黄芪提取物各剂量组、抗疲劳配方各剂量组、黄芪复合配方各剂量组能够升高血清乳酸脱氢酶含量(P＜0.01,P＜0.001);黄芪提取物高剂量组和抗疲劳配方中、高剂量组以及黄芪复合配方各剂量组尿素氮水平显著降低(P＜0.05,P＜0.01);黄芪提取物高剂量组、抗疲劳配方各剂量组和黄芪复合配方各剂量组Na+-K+-ATP酶水平显著升高(P＜0.05,P＜0.01).与空白组相比,黄芪复合配方中剂量组上调差异代谢物有69个,下调差异代谢物有149个.黄芪复合配方组与空白组对比差异代谢通路有咖啡因代谢、甘氨酸、丝氨酸和苏氨酸的代谢、苯丙氨酸、酪氨酸和色氨酸的生物合成等.结论 黄芪复合配方可发挥更好的抗疲劳功效.代谢组学结果表明,黄芪复合配方可能通过调控咖啡因代谢通路和体内氨基酸水平延缓疲劳的产生、提高供能,进而发挥抗疲劳作用.
Research on anti-fatigue effect and metabolomics mechanism of Astragalus composite formula
Objective To elucidate the anti-fatigue effect and metabolomics mechanism of the Astragalus composite formula.Methods To elucidate the anti-fatigue effect and metabolomics mechanism of Astragalus composite formula.Methods Male Kunming mice were randomly divided into 10 groups,namely,blank group,low,medium and high dose group of Astragalus extract,low,medium and high dose group of anti-fatigue formula,low,medium and high dose group of Astragalus composite formula,and each group was subjected to the weight-bearing swimming experiment after 29 days of continuous gavage adminis-tration.After 30 days of continuous gavage,mice were sampled after weight-bearing swimming,and serum was separated to detect urea nitrogen,lactic acid,and lactate dehydrogenase;muscle was separated to detect myoglycogen,Na+-K+-ATPase,and Ca2+-Mg2+-ATPase;and the skeletal muscles of 6 mice from each of the blank group and the middle-dose group of Astragalus composite formula were randomly selected,and the skeletal muscles of 6 mice from each of the blank group and the middle-dose group of the Astragalus composite formula were examined by ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry.Metabolomics by ultra-high performance liquid chromatography coupled to quadrupole time-of-flight mass spectrometry(UHPLC-QTOFMS),PCA-QTOFMS and PCA-QTOFMS.QTOFMS),PCA and OPLS-DA multivariate statistical analysis methods to screen out the endogenous differential metabolites of the Astragalus composite formula for anti-fatigue,to identify the metabolic pathways associated with the differential metabolites,and to investigate the mechanism of action of this Astragalus composite formula for the alleviation of exercise fatigue.Results After the mice in each group were given samples,the results of weight-bearing swimming experiments showed that compared with the blank group,the exhaustion swimming time of mice in each dose group of Astragalus extract,each dose group of anti-fatigue formula and each dose group of Astragalus compound formula was significantly prolonged(P＜0.05,P＜0.001);the detection of serum and muscle indexes showed that compared with the blank group,the muscle glycogen content was significantly elevated(P＜0.05,P＜0.01);blood lactate was significantly reduced in all groups except Astragalus extract group(P＜0.05,P＜0.01,P＜0.001);Astragalus extract dose groups,anti-fatigue formula dose groups and Astragalus composite formula dose groups were able to elevate the serum lactate dehydrogenase content(P＜0.01,P＜0.001);Astragalus extract high dose group and anti-fatigue formula middle and high dose groups as well as Astragalus composite formula each dose group significantly decreased urea nitrogen level(P＜0.05,P＜0.01);Astragalus extract high dose group,anti-fatigue formula each dose group and Astragalus composite formula each dose group significantly increased Na+-K+-ATPase level(P＜0.05,P＜0.01).Compared with the blank group,there were 69 up-regulated differential metabolites and 149 down-regulated differential metabolites in the Astragalus composite formula medium dose group.Differential metabolic pathways in the Astragalus Complex Formula group compared with the blank group were caffeine metabolism,metabolism of glycine,serine and threonine,and biosynthesis of phenylalanine,tyrosine and tryptophan.Conclusion Astragalus composite formulation may exert better anti-fatigue efficacy.The metabolomics results suggest that the Astragalus composite formula may delay the production of fatigue and improve energy supply by reg-ulating the caffeine metabolic pathway and amino acid level in the body,and then exerting anti-fatigue effects.
Astraglussupplement qi and fortify the spleenantifatigueenergy metabolismmetabolomics
万方数据
维普
