An Experimental Study On Anti-fatigue Effects And Relevant Mechanism Of Plants

ObjectiveTo establish an animal loaded swimming model,comparing the antifatigue effects of several substances.To investigate the effects of extraction on anti-fatigue metabolism in swimming mice and the involved mechanisms related to energy metabolism,antioxidant system and the expression of liver proteins.This study will provide a new way for the development of anti-fatigue supplements in the future.Methods1.To Compare the anti-fatigue effect of several substances:mice were fed with maltodextrin,ginseng leaves extract,and Moringa oleifera leaves for 14 days.By recording the swimming time with a load on their tails equivalent to 2%body weight to select the best antifatigue substance.The fatigue swimming test was performed on mice by orally taking the Moringa leaves 500 mg/kg,1000 mg/kg and 2000 mg/kg dose to determine the best gavage dose for the next intervention.2.To Analyze the main components of the extract of Moringa leaves:The fresh Moringa oleifera leaves were washed,dried,broken,sieved,and the Moringa leaf powder is extracted with a solvent.Then the extract was filtered,concentrated in vacuo,and freeze-dried to get a tan powder.Exhaustive swimming time of the mouse was used as an evaluation index for the extraction effectiveness.The total antioxidant capacity,total phenolics,plant flavonoids and polysaccharides content of the extract Moringa oleifera leaves were determined by FRAP,Folin-Phenol,Al-Ion color method and phenol-sulfuric acid method,and the possible toxicity of the extracts was examined by acute toxicity test.3.Preliminary study on the mechanism of anti-fatigue and anti-oxidation in the extract of Moringa oleifera leaves:To set three groups:The normal control,swimming control and EML（extract of Moringa leaves）group.The mice were fed for 14 days with 90min swimming.Then the blood was taken from the retro-orbital venous plexus,liver and gastrocnemius muscles were collected from mice after cervical dislocation.Blood glucose,lactic acid（Lac）,urea nitrogen（BUN）,and glutathione（GSH）,Tissue glycogen and ATPase were measured.The Plasma and tissues contents of Malondialdehyde（MDA）,total antioxidant capacity（T-SOD）,glutathione peroxidase（GSH-Px）,catalase（CAT）,lactate dehydrogenase（LDH）and free fatty acids（NEFA）were assayed,respectively.The liver ORM1 and CCR5 were quantitatively measured by the Enzyme-linked immunosorbent assay（ELISA）.Results1.Moringa oleifera leaves could significantly prolong the swimming time（P<0.05）after loaded swim,compared with the control group,and the time was higher than the extract of ginseng leaves group（positive control）.Moreover,medium dose group（1000 mg/kg）has a good effect of improving exercise tolerance.2.The yield of Moringa oleifera leaf extract was（32.37±1.71）%.Compared with Moringa leaves material,the EML was found to notablely increase the swimming time by 19.27%（p<0.05）.The RML could not significantly raise the exercise time in mice,compared with the control group.By analyzing the components of the EML and RML,the obtained data showed that the active ingredients in the RML were very low,and the total antioxidant capacity of EML is 137 times higher than the RML and the plant flavonoid content is 1211 times,and the total polyphenol content is 129 times,respectively.The LD₅₀ of leaf extract was more than10g/kg.3.Supplementation of EML has a significant effect on both energy regulation and antioxidant systems.The EML had no diverse effect on the organ weight in mice.Among the serum markers,there were statistical differences in the four indicators of the Moringa oleifera extract group（P<0.05）versus the swimming control,in which the content of BUN in the EML was significantly lower than that of the swimming control（P<0.01）.Both muscle and hepatic glycogen in groups were significantly consumed after exercise（P<0.05）,the level of glycogen were improved by Supplementation of EML（P<0.05）,but they did not reach the normal at rest.The activity of Na⁺K⁺-ATPase in the liver was significantly higher than that in the swimming control group（P<0.05）.After applying the EML,the activity of SOD and CAT increased,and GSH-Px in the liver had a obviously raised trend（P<0.01）.LDH had a specific distribution,and its activity in plasma was statistically significant,but it was not reflected in tissue.MDA content in serum and liver tissue is apparently lower than that in the swimming control group.With the occurrence of exercise,the expression of ORM1 in liver tissue increased significantly,and the expression of the transduction receptor CCR5 also had the same change.The reason was that exercise promotes lipolysis,and the EML participates in lipid mobilization.It also regulated glucose metabolism,promote glycogen synthesis and energy supply,and had a protective effect against oxidative stress.ConclusionThis study found that the Moringa oleifera had a good anti-fatigue effect.Its crude extract had revelant anti-oxidative components and played an important role in energy metabolism regulation and antioxidatant amelioration.This study will provide a new idea and approach for the development of anti-fatigue functional foods.