| Objective:This paper aims to observe the intervention effect of selenium enriched tea polysaccharide on the regulating effect of fatigue through the establishment of exercise fatigue and chronic fatigue model. We analysis the change of endogenous metabolites in chronic fatigue rat intervened by selenium enriched tea polysaccharide by using GC-TOF-MS metabonomics technology combined with the multi-dimensional statistical means, and confirm the different related metabolic markers and metabolic pathways, as well as explore the changes of metabolic network in rats with chronic fatigue and the metabolic mechanism of the prognosis of stem polysaccharide enriched tea, which can provide the possible experiment basis for the prevention and alleviation of the CFS.Methods:(1) Material preparation and the physical and chemical propertiesPrepare Se-TP by water extraction and alcohol precipitation, then detect the structure of purified Se-TP. Detection contents include:polysaccharide content, selenium content, protein content, molecular weight, composition of sugars, amino acid composition, ultraviolet spectrum, infrared spectrum and environmental scanning electron microscope.(2) Delay effect of sports fatigue by taking Se-TPEstablish a 2 week mouse loaded swimming training model, which would suffer intragastric administration of different doses of selenium enriched tea polysaccharide in aqueous solution, and measure blood glucose, blood lactate (Bla), blood urea nitrogen (BUN) and liver glycogen, glutathione peroxidase (GSH-Px) and malondialdehyde (MDA) content at the moment of exhaustion of strength and exhaustion of 24h. Discuss the effect of selenium enriched tea polysaccharide on sport fatigue(3) The improvement of Se-TP for chronic fatigueEstablish a chronic fatigue model by using the constraint+running platform+ sleep disorder+congestion+noise induced by multiple factors. Assess the impact of CFS model and selenium tea polysaccharide on CFS through the observation of rat body weight, intake of food and drinking water quantity, combined with general scoring scale and behavioral science experiment (water maze, open field, hanging tail test).(4) Metabolite screening of chronic fatigue by taking Se-TPFind out endogenous metabolic markers induced by chronic fatigue rats and seleno teapolysaccharide through GC-TOF-MS Technology of metabolism. Seek the potential differences in metabolites by the multivariate statistical methods and principal component analysis and orthogonal partial least squares. Identify Potential biomarkers by the KEGG database which would be entered to MetPA database for the construction and analysis of the corresponding metabolic pathwayResults:(1) Se-TP got 3 peaks after the DEAE-52 cellulose separation, of which the peak2 yield up to 53.69%; Se-TP is a single symmetrical peak after purification by G-150 Sephadex column. Se-TP molecular weight is 150kda, composed of mannose, rhamnose, glucose, galactose, arabinose and galacturonic acid. The amino acid composition are mainly aspartic acid, serine, glutamic acid, alanine, cysteine amino acid, valine, isoleucine, leucine, lysine lysine, histidine and arginine acid. Ultraviolet spectrometer scanning analysis found out:Se-TP exist a distinct protein absorption peak in 285.5nm and infrared spectrum showed Se-TP belongs to the acidic polysaccharides. Environmental scanning electron microscope showed that the morphology of Se-TP was relatively smooth, and the shape was more regular and the size was uniform.(2) Serum Bla,BUN concentration and liver MDA concentration increased, hepatic glycogen content and GSH-Px activity decreased after exhaustive of mice; Whether exhausted immediately or after 24h recovery, Se-TP could significantly prolong the exhaustive swimming time of mice, reduce the concentration of Bla and BUN, increase the activity of GSH-Px, reduce the liver MDA concentration, and accelerate the recovery of liver glycogen.(3) Chronic fatigue rat model validation results:CFS rats appeared depressed, back arched; skin relaxation, the hair withered; Irritability, aggression, and reduced activity; The weight, daily feed intake, tail suspension struggling times and still of CFS rats significantly reduced, modified number increased, vertical number and the times of crossing lattice reduced, escape latency only increased significantly the next day, times across the platform quadrant was significantly lower than that of group C. Compared with the CFS group, resting time of Se-TP intervention group were significantly reduced, struggling times was lower, it had no significant difference; the escape latency shortened, the number of cross platform quadrant significantly increased; wearing lattice times increased the vertical number and time modified number seemed no significant difference compared with CFS group(4)Metabolome analysis showed that urine after exercise produced 13 differential metabolites in CFS group compared to C group,6 metabolites significantly increased: malic acid, fumaric acid, palmitic acid, Kang acid, amino malonic acid, sphingosine; 7 metabolites significantly decreased:a-Ketoglutaric acid, Glucose acid, Tartaric acid, malonic acid,2-amino-l-phenylethanol,21-Hydroxypregnenolone. There were 8 different metabolites in urine after exercise in Se-TP group compared to CFS group, of which benzoic acid was downtrend, Creatine, itaconic acid, glutaric acid, 21-Hydroxypregnenolone,2-isopropyl malic acid, levodopa and gamma aminobutyric acid were upward trend. There are three metabolic system that Se-TP influence on CFS, according to:tyrosine metabolism system, steroid hormone metabolism and arginine proline metabolism system.Conclusions:(1) Ultraviolet visible spectrum scanning analysis showed that Se-TP belongs to the protein polysaccharide.Infrared spectrum analysis indicated Se-TP is sugar ring, which is a kind of acidic sugar. HPLC points out Se-TP is an acidic polysaccharide containing a high sugar acid. Environmental scanning electron microscope showed that the morphology of Se-TP was smooth, the shape was normal.(2) In vitro antioxidant activity of Se-TP showed Whether it was in the exercise of immediate or 24h recovery, the supplement Se-TP could prolong the time of exhaustive swimming in mice, reduce the concentration of BUN, and Bla, improve the activity of GSH-Px, decrease the content of MDA, and accelerate the recovery of liver glycogen, which would protect the body against Lipid peroxidation caused by sports fatigue.(3) The chronic fatigue model was successfully established by the method of the multiple factors combined with the constraint and the crowded+noise and the sleep disorder. Supplement of Se-TP had no significant effect on body weight and water content in rats, markedly affected physical characteristics, the tail suspension immobility time, open field cross frequency, escape latency and crossing times, relieved the fatigue and depression of rats, improved spatial memory ability, and had certain improvement function to the chronic fatigue.(4) The experimental results show that CFS1 VS C1 and F v CFS1 group have the common metabolites:y-aminobutyric acid and 21-hydroxy pregnenolone.It involves common metabolic pathways that is steroid hormone metabolism, it could be the effect of chronic fatigue syndrome pathways by taking the Se-TP. |
PDF Full Text Request