| Obejective:By establishing the rat model with exercise-fatigue and chronic fatigue syndrome, We explore the influences of this two type of fatigue on the organism and the improve effect of Schisandra chinensls Polysaccharide (SCP)on this two type of fatigue. Moreover, the impact of exercise-fatigue and chronic fatigue syndrome for exercise capacity was clarified, provided an experiment basis for the SCP.Methods:1) The polysaccharide of Schisandra chinensls was obtained through hot water extraction and ethanol precipitation, then purified with DEAE-cellulose column and Sephadex column and study its physic-chemical property, which include the content of glycogen and protein, UV and IR, monosaccharide composition analysis and amino acids analysis, then using environmental scanning electron microscopy and atomic force microscope to observe the molecules of SCP.2) Swimming training for five weeks in mice was used to build a sports fatigue model and the control group was established to study the effect of SCP on motor fatigue. Indexes of anti fatigue test include body weight, exhaustive swimming time and biochemical indexes(Blood lactic acid, liver glycogen, muscle glycogen, CK,BUN)3) CFS animal model was constructed in 4 weeks. Experiment entered the stage of treatment after the model was successfully. Test the index of animals' behavior and collect the urine of animals after exhaustive exercise. Comparison and identification of animal urine samples using GC-TOF-MS technique. The indexes of animal behavior were body weight, food and water intake, water maze test, the tail suspension test, open field test. Urinary metabolic parameters were screening and identification of urine routine indexes, total ion map analysis, PCA analysis, OLPS-DA analysis and differential metabolites.Results:1) The contents of sugar was 58%. The UV showed that SCP belongs to a kind of proteoglycan, the IR indicated that SCP belongs to a-configuration. During monosaccharide composition analysis experiment, SCP contains mannose, glucose and galactose; the result of amino acids content test shows that SCP have 12 amino acids, include Ala, Tyr, Glu, Phe, Val, Arg, Lys, His, Ser, Ile, Gly, Asp, total protein of SCP 1 was 4.53%. Environmental Scanning Electron Microscopy and atomic force microscopy showing SCP1 is make up of small irregular particles and the solid amorphous structure.2) The study of SCP on anti-exercise-fatigue, SCP high-dose group rats' exhaustive swimming time longer than the control group, with a significantly different (P<0.05), indicates that the high-dose of SCP has a delayed effects on exhaustive fatigue time. At the time of exhaustion, the content of blood lactic acid?CK and BUN in SCP low-dose group and high-dose group reduced than the control goup (P<0.05, P<0.01). There were no significant difference on liver glycogen and muscle glycogen (P>0.05). The comparison in each group, after 24h the content of blood lactic acid, liver glycogen, muscle glycogen and CK were recovered, and the difference was significant (P<0.05, P<0.01).3) To study the improve effect of SCP on chronic fatigue syndrome and metabolic mechanism, CFS model was established. Result shows that the CFS group rats'listless, and skin relaxation phenomenon has a significant difference compared with the control group (P<0.05), the SCP rats score decreased (higher scores, worse the mental health), and has a significant difference(P<0.05). Group CFS rats'daily food intake and weight gain were significantly lower than the control group (P<0.05), and no significant difference in the amount of drinking water (P>0.05), the SCP group's food intake and body weight growth rate was significantly higher than that of the control group (P<0.05), and the amount of water had no significant difference (P>0.05). In water maze test, the time of SCP model rat's finding the original platform location was longer than CFS group (P<0.05). In the field experiment, the rats in CFS group by distance, speed, stand number were significantly lower than those control group (P<0.05, P<0.01), the modification times higher than that of control group (P<0.05), the SCP group at the four quarry in the experiment (total distance, average speed, the number of standing and modification times) showed significant difference with CES group(P<0.05). In the tail suspension test, no significant difference in struggling times during the three groups of rats (P>0.05), but the CFS group's static statues was higher than that of control group (P<0.05), and the SCP group lower than that of CFS group (P<0.05). Collect the urine of the three groups rats after exhausted exercise, and then test these sample, two-dimensional map of PCA showed quiet group and CFS group urine sample separation trend is obvious, and there are some overlapping phenomenon, the CFS group and SCP group sample separation trend was significant, and no overlapping phenomenon, the three-dimensional shows the result that each group were separated obviously. The final screening and identification of metabolic difference results show that the difference were substation between CFS and control group:Fumaric acid, 2-methylfumarate, Aminomalonic acid, L-Malic acid, alpha-ketoglutaric acid, tartaric acid,2-Amino-l-phenylethanol, gluconic acid 1 and palmitic acid. the difference substation between SCP and CFS group were:pimelic acid, Aminomalonic acid, L-Malic acid, Creatine and benzoic acid. These compounds are mainly involved in Glyoxylate and dicarboxylate metabolism, Glycine, serine and threonine metabolism, Phenylalanine metabolism, Alanine, aspartate and glutamate metabolism and so on.Conclusion:1) The SCP can reduce the content of lactic acid after the moment of exercise, accelerated muscle and liver glycogen recovery rate. It has obvious anti fatigue effect, but also can reduce the serum CK activity, resist BUN, relieve fatigue, adding the exhaustive swimming time of mice.2) The supply of SCP can increase the weight of rats, space exploration ability, memory and motor ability, alleviate the symptoms of Chronic Fatigue Syndrome.3) By the technology of GC-TOF-MC, SCP can improve CFS and may through the compound of L-Malic, Fumaric acid to adjust TCA, Alanine, Aspartic acid, Glutamic metabolism. This study has an enlightening significance to prevent and improve the mechanism of chronic fatigue syndrome of the human body, but it needs to be further studied. |
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