| Brassica rapa L.is a three-purpose crop in the Qinghai-Tibet Plateau,which is used for medicine,food and feeding.It has a long history of planting.It is recorded in the classic Tibetan medicine “Four Medical Classics” that Brassica rapa L.root has the functions of clearing away heat and detoxifying,nourishing and increasing oxygen.Modern medicine shows that its significant pharmacological effects of anti-oxidation,anti-altitude hypoxia and etc.,but there is no in-depth research on its anti-fatigue effect and mechanism.Fatigue affects people’s physical and mental health,and even threatens life.With the accelerated pace of society,the incidence of fatigue is getting higher and higher and the development of natural products with high antifatigue activity has become a research hotspot.Therefore,this study took the water extract of Brassica rapa L.as the research object,characterized its chemical components,predicted the possible anti-fatigue mechanism using network pharmacology,and used the weight-loaded swimming mice to verify its function.Based on serum metabolomics,we further analyzed Coriander Mechanisms of anti-fatigue action of roots.A total of 46 components were identified after qualitative analysis of the chemical components of the water extract of Brassica rapa L.by UPLC-MS method,including 9 kinds of organic acids,9 kinds of esters,8 kinds of glycosides,8 kinds of glucosinolates,5 kinds of flavonoids,2 kinds of phenolics and other compounds.Based on the results,combined with the active ingredients screened in the database,the potential targets were predicted,and an “active ingredient-targets” network of 33 anti-fatigue active ingredients and 114 potential gene targets was constructed.Cortexin,trans-6-gingerol and 6-gingerol are the important components of the water extract of Brassica rapa L.The results of GO(Gene Ontology)biological process analysis showed that Brassica rapa L.played an anti-fatigue effect mainly by regulating protein phosphorylation,ATP binding,and regulating protein kinase activity.The main signaling pathways involved in root anti-fatigue targets are PI3K-Akt,MAPK,m TOR,insulin signaling pathway,TNF,AMPK and other signaling pathways,indicating that Brassica rapa L.exerts anti-fatigue effects via multi-components,multi-targets and multi-ways.The anti-fatigue effect of the water extract of Brassica rapa L.was verified by mouse behavioral experiments and weight-loaded swimming test,and its mechanism of action was explored through serum metabolomics.The water extract of Brassica rapa L.can significantly improve the forelimb grip and exercise time on the rota-rod test,showing the enhancement of exercise performance and endurance.In terms of relieving exercise-induced fatigue caused by weight-bearing swimming,the water extract of Brassica rapa L.can significantly reduce the accumulation of metabolites lactate and urea nitrogen in serum,and improve the activity of lactate dehydrogenase in muscles.The measurement results of energy metabolism-related indicators showed that the water extract of Brassica rapa L.significantly increased the levels of glycogen reserves and ATP,as well as the levels of key enzymes in energy metabolism,succinate dehydrogenase and pyruvate kinase,and regulated energy metabolism.Relieve muscle damage caused by strenuous exercise.In addition,the water extract of Brassica rapa L.can significantly enhance the activity of antioxidant enzymes in liver,and reduce the accumulation of lipid peroxidation product malondialdehyde.On the other hand,coriander root has a certain anti-inflammatory effect,and probably plays a key role in regulating central neurotransmitters,delaying the occurrence of exercise-induced central fatigue.The RT-PCR and Western blot analysis showed that the water extract of Brassica rapa L.can activate the AMPK/PGC-1α signaling pathway,and at the same time increase the m RNA and protein expression levels of Nrf2 and HO-1 in the liver of mice.Serum metabolomics research method was used to further study the anti-fatigue mechanism of Brassica rapa L.The differential metabolites screened were choline,acetoacetic acid,taurine,nicotinamide,lactic acid,etc.The results of metabolic pathway analysis showed that linoleic acid was mainly involved.Metabolism,the generation and degradation of ketone bodies,taurine and hypotaurine metabolism,and niacin and nicotinamide metabolism are closely related to the regulation of energy metabolism.Finally,a coriander root compound product “Yuanqi Yangyuan Decoction” composed of Brassica rapa L.,astragalus root and tangerine peel was developed,and it was verified that the water extract of this compound preparation has a synergistic effect and its anti-fatigue effect is better than that of coriander root.water extract. |
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