Extraction And Purification Of Flavonoids From Moringa Oleifera Lam.Leaves And Their Anti-fatigue Activity

ObjectiveMoringa oleifera Lam.is a perennial tropical subtropical deciduous plant.Moringa oleifera Lam.leaves are rich in nutrients and functional ingredients,and flavonoids are one of the main functional ingredients.The flavonoids of Moringa oleifera Lam.leaves can prevent diseases such as hypertension and hyperlipidemia,and play an antioxidant and immune-improving role.So far,anti-fatigue efficacy of them have not been reported in the literature.This project mainly separated and purificated flavonoids from Moringa oleifera Lam.leaves,studying its anti-fatigue activity and molecular mechanism.All these provide theoretical support for the research of Moringa oleifera Lam.leaves flavonoids.in functional food.Methods1.Through the investigation of the static adsorption and desorption effects of AB-8,NKA-?,D101,HPD600,S-8,ADS-7,X-5 and DM301 macroporous resins on the crude extract of Moringa oleifera Lam.leaves,an optimal resin was selected to use for the separation and purification of flavonoids,and its static and dynamic adsorption and desorption conditions were optimized to obtain the optimal purification process conditions.The flavonoids content in the extracts before and after purification was compared.Analysis of flavonoids in the purification material was performed by HPLC.2.The anti-fatigue activity of MLFE was explored.Forty-eight male Kunming mice were divided into normal control(NC)group,Moringa oleifera Lam.leaves crude extract(MLCE)group and Moringa oleifera Lam.leaves flavonoids extract(MLFE)group.The NC group was orally administered pure water,and the MLCE group and the MLFE group were orally administered with the solution of Moringa oleifera Lam.leaves crude extract(340 mg/kg·bw)and Moringa oleifera Lam.leaves flavonoid extract(75 mg/kg·bw)0.5 m L,D15 weight-bearing swimming,record exhaustive swimming time;48 mice were divided into quiet control(QC)group,swimming control(SC)group and Moringa oleifera Lam.leaves flavonoids extract(MLFE)group.QC and SC group were orally administered pure water,the MLFE group was orally administered the Moringa oleifera Lam.leaves flavonoids extract solution.The perfusion volume was 0.5 m L.D15.The SC and MLFE groups were subjected to non-weight-bearing swimming for 90 min.Blood,liver,and hind leg muscles were taken.Blood was taken directly from the stomach and then sacrificed.Liver and hind leg muscles were taken to determine relevant biochemical indicators.3.Paraffin-embedded sections were used to prepare pathological sections for histopathological observation;q-PCR,Elisa and other experimental techniques were used to investigate the m RNA of GCK/PEPCK,Nrf-2/HO-1 and other related genes in muscle after stress and intervention and protein level expression.Result1.The optimum process conditions of NKA-? macroporous resin for MLFE separation and purification are obtained: ambient temperature 25?,loading flow rate 1.5 BV/h,The loading concentration was 1.12 mg/m L,the loading volume was 5 BV,the ethanol concentration was 70%,the elution flow rate was 2.5 BV/h,and the eluent volume was 11 BV.The flavonoids content in the sample solution increased from 12.36% to 54.66%.2.An experimental model of exhaustive swimming in mice was found to show that MLFE significantly prolonged the exhaustive swimming time of mice,and the effect was slightly better than MLCE.It also increased Glu of mice after swimming,and reduced the levels of LA,BUN,and MDA.Maintaining muscle liver glycogen levels during exercise,increasing the activity of antioxidant enzymes,ATP-ase and NEFA in serum and tissues.3.The flavonoids of Moringa oleifera Lam.leaves can enhance the expression of GCK,PEPCK,Nrf-2,HO-1 genes and proteins.Conclusion1.Purification of MLFE by using NKA-? macroporous resin has greatly improved the purity of flavonoids,from 12.36% to 54.66%,which proves that the purification process conditions are feasible.2.In mouse swimming experiments,MLFE can enhance swimming endurance of mice,increasing the levels of Glu,muscle liver glycogen,and NEFA,reducing the levels of LA,BUN and MDA,and promote body energy metabolic speed;changing the way of energy metabolism;increasing ATP-ase and various antioxidant enzymes activity,and maintaining the balance of oxidative antioxidant system in the body.MLFE achieve antifatigue effects by regulating energy metabolism,reducing accumulation of metabolites,and improving antioxidant capacity.3.It was found that MLFE can play an anti-fatigue role by regulating the GCK/PEPCK,Nrf-2/HO-1 signaling pathways,regulating energy metabolism and antioxidant capacity.