| Oxidative stress is a negative effect of free radicals in the body,and is the main factor of disease occurrence and development.Modern animal husbandry is developing towards large-scale and intensive,which leads to oxidative stress becoming a problem that must be faced in the process of livestock and poultry breeding.The development of natural antioxidant active ingredients to improve the antioxidant capacity of livestock and poultry has become a key research direction in the livestock industry,and the rich flavonoids in Acanthopanax,which have significant antioxidant activity,have received extensive attention from domestic and foreign researchers and scholars.Therefore,it is of great importance to optimize the extraction process of the total flavonoids of Acanthopanax,and to study its antioxidant activity and mechanism of action.Optimization of the extraction process of total flavonoids of Acanthopanax.Ultrasonic-assisted ethanol extraction of the total flavonoids of Acanthopanax,the effect of different factors on the extraction rate of total flavonoids was analyzed,and response surface tests were designed to optimize the extraction conditions.The results show that the highest extraction rate of total flavonoids(24.11±0.17 mg·g-1)was obtained when the ethanol concentration was 55%,the extraction time was 73 min,the liquid to material ratio was 1:45g·m L-1,the extraction temperature was 72°C and extraction power was 300 W.Evaluation of antioxidant activity of total flavonoids from Acanthopanax senticosus in vitro.Determination of the scavenging ability of total flavonoids of Acanthopanax against DPPH radicals,ABTS radicals,hydroxyl radicals,and total reducing ability.The results showed that the IC50 values of the scavenging ability of the total flavonoids of Acanthopanax on DPPH radicals,ABTS radicals and hydroxyl radicals were 28.38μg·m L-1,107.77μg·m L-1 and 228.70μg·m L-1,respectively,and the total reducing ability was positively correlated with the concentration.Protective effect of total flavonoids of Acanthopanax on oxidative stress in RAW264.7cells.Establishment oxidative stress modeling by H2O2 stimulation of RAW264.7 cells.To investigate the effects of total flavonoids of Acanthopanax on on ROS level,MDA content,and the activities of CAT,SOD and GPx.To detect the expression levels of Nrf2,Keap1 and HO-1 proteins by Western Blot.The results showed that the total flavonoids of Acanthopanax can promote the proliferation of RAW264.7 cells,can reduce the ROS level and MDA content in the cells,and increase the CAT,SOD and GPx activities.Meanwhile,it was also able to exert inhibitory effects on Keap1 protein,promote the transfer of Nrf2protein into the nucleus and increase the level of HO-1 protein.Protective effect of total flavonoids of Acanthopanax on ulcerative colitis in C57BL/6mice.Ulcerative colitis model in C57BL/6 mice induced by drinking 2.5%sodium dextran sulfate for 7 days.Preliminary investigation on the mechanism of action of total flavonoids of Acanthopanax in inhibiting ulcerative colitis by increasing antioxidant activity in mice.The results showed that total flavonoids of Acanthopanax can effectively prevent colon length shortening and tissue damage,reduce disease activity index and histopathological score in mice,and can improve CAT,SOD and GPx activities and reduce MDA content in colon tissues.The experiment was conducted to optimize the extraction process of total flavonoids of Acanthopanax,which improved the extraction rate of total flavonoids of Acanthopanax and was of great significance to the conservation of Acanthopanax resources.In vitro free radical scavenging assay demonstrated the strong in vitro antioxidant activity of total flavonoids ofAcanthopanax.Total flavonoids of Acanthopanax can reduce intracellular ROS levels and inhibit lipid peroxidation by activate the Nrf2/Keap1-HO-1 signaling pathway,which have strong protective effects on macrophages under oxidative stress.Total flavonoids of Acanthopanax can inhibit ulcerative colitis by increasing the antioxidant activity in animals.In vitro and in vivo antioxidant experiments are of reference significance for the development and application of the products related to the total flavonoids of Acanthopanax. |
