Study On Cell Distruption And Anti-Oxidant Of Bee Pollen In Vitro And In Vivo

Bee Pollen is an irregular,oblate spheroid particle formed by mixing pollen balls collected from the anthers of pollen-derived plants with their own glandular secretions,nectar and saliva.The surface of bee pollen is surrounded by bee pollen wall and its main component is sporopollenin,which is acid-resistant,alkali-resistant and corrosion-resistant.Itcan effectively protect pollen from external aggression.Bee pollen is rich in nutrients,but the bee pollen wall hinders the dissolution of the nutrients in the bee pollen and the human body's full use of the nutrient components of the bee pollen.China is a big producer of bee products.The annual trade volume of bee pollen exceeds 5,000 tons.However,the processing rate is low,and the comprehensive value of bee pollen cannot be realized.Therefore,selecting a suitable wall-breaking process to effectively break the wall of the bee pollen to make the nutrient flow out is beneficial to the body's full absorption of nutrients in the bee pollen,and it is beneficial to the processing and comprehensive utilization of bee pollen.In this paper,different kinds of bee pollen were selected,and the corresponding bee pollen was treated with ball milling or ultrasonic wall breaking.By comparing the morphological changes before and after wall disruption of bee pollen,the nutrients dissolved out,the in vitro antioxidant activity changed.The liquid chromatography was used to identify the chemicals in the ethanol extracts.And rose bee pollen was chosen to evaluate the anti-oxidation properties of the digested products in vitro simulated digestion.The in vivo anti-aging activity was evaluated using the aged mouse model and the changes in nutrient composition and inantioxidant activity of bee pollen after the wall breaking treatment was systematically evaluated.The main conclusions are as follows:1)The particle size of bee pollen was analyzed by laser particle size analyzer,the morphological changes of bee pollen were observed by scanning electron microscopy,and the protein,crude fat,soluble sugar,free amino acids,total flavonoids,vitamin B1,vitamin C and free fatty acid content in bee pollen were determined by the corresponding food analysis methods.The results showed that both ball milling and ultrasonic disruption could reduce the particle size of bee pollen and make its outer wall completely open.Both methods could effectively increase the content of protein,crude fat,total flavonoids,free amino acids,and some soluble sugars and free fatty acids in bee pollen.But it can reduce the contents of vitamin B1,vitamin C,soluble sugar and free fatty acid species.2)ORAC,DPPH and ABTS were used to evaluate the in vitro antioxidant activity of different varieties of bee pollen.Person analysis was used to analyze the correlation coefficient between the content of each nutrient component and the antioxidant activity,liquid chromatography and mass spectrometry were used to compare ethanol extracts of different kinds of bee pollen and thestructure was identified.The results showed that the cell disruption treatment could significantly increase the antioxidant activity of bee pollen in vitro.The positive correlation between antioxidant activity and total flavonoid content in vitro was the strongest,and thestrongest negative correlation was particle size.Among them,rose bee pollen had strong antioxidant activity and smallest particle size;the ethanol extracts of bee pollen are mainly flavonoid glycoside compounds and spermidine derivatives,and the flavonoid glycosides are mainly composed of isorhamnetin,kaempferol and quercetin,and glucoside,rhamnoside or neohesperidoside.The spermine derivatives are mainly composed of spermidine and caffeoyl or feruloyl.Among them,the spermidine derivatives in rose bee pollen are rich in species and have higher contents.3)Rose bee pollen was chosen as the representative.In vitro simulated digestion experiments was used and ORAC,DPPH,and ABTS methods were used to evaluate the changes in the antioxidant activity ofthe digestiveof bee pollen during in vitro digestion;a 12-month old mouse model was used.The oxidation-associated enzymes and metabolites of SOD,LDH,CAT and MDA in mouse serum were measured to evaluate the changes in the antioxidant activity of bee pollen.The results of in vitro simulated digestion showed that after the wall-breaking treatment,the antioxidant capacity of rose bee pollen juice was significantly increased,and the intestinal fluid removal ABTS ability was also significantly enhanced,but there was no significant change in the ORAC value and DPPH value of intestinal fluid;antioxidant results in vivo showed that the broken rose bee pollen could significantly increase the content of SOD,LDH and CAT in serum of old rats and significantly reduce the content of MDA.In conclusion,the wall-disrupting treatment can effectively improve the antioxidant activity of bee pollen in vitro and in vivo.In summary,after cell disruption treatment of bee pollen,many nutrients are dissolved,which not only enhances the antioxidant activity of bee pollen in vitro,but also improves the antioxidant activity of bee pollen in vivo,thereby improving the physiological environment and delaying aging in vivo.