The Comparative Analysis Of Construction And Bioactivity Of Self-assembled Micelles Loaded With DHA From Different Food Proteins

DHA belongs to an n-3 series of unsaturated fatty acids with good antioxidant,lipid-lowering and anticancer effects.However,DHA is easily oxidized and decomposed during processing and storage,and is easily inactivated under light and high temperature.How to keep DHA active has become an urgent technical problem.This study selected ovalbumin micelle,myosin micelle,soybean globulin micelle and?-milk protease hydrolysis of protein micelles encapsulates DHA,part four protein after chymotrypsin enzyme solution formed amphiphilic peptides,DHA package will be buried in the micelle interior,through exploring the micelle structure characteristic and the influence on biological activity of DHA,the four kinds of DHA protein micelles for comparative analysis.The results show that:The secondary structure and fluorescence quenching of four proteasolytic proteins were determined by fluorescence spectrum and circular dichroism.The electrokinetic potential of DHA-lactoglobulin micelles was-24.3 mV,and the structure system was the most stable.DHA loading soybean globular protein micellar tractor-helix decreased by 2.6%,and DHA encapsulation rate and embedding volume were the highest,which were 91.53%and 196,respectively.The particle size of DHA-carrying myosin was 23 nm.Ovalbumin micelles had an electric potential of-18.3,an average particle size of 38nm,and a embedding capacity of185.The stability and in vitro antioxidant properties of the delivery system were further determined.The DHA loaded lactoglobulin micelles were the most water-soluble and stable,with the retention rate of 34.83%under high temperature and 36.42%under ultraviolet irradiation,and the best antioxidant properties.The scavenging rates of ABTS and DPPH free radicals were 38.85%and 48.44%,respectively,and the Vc equivalent was 49.53 g/mL.The best light stability and thermal stability of docosaenoglobin micelles were 36.71%and37.53%,respectively.Docosahexaenoalbumin micelles showed good antioxidant activity,with Vc equivalent of 42.92 g/mL.By using HepG2 cells to establish a high-fat model to detect the lipid-lowering function of cells,four kinds of DHA carrying micelles showed good membrane pass rate,and the transport speed of DHA carrying ovalbumin micelles,DHA carrying myosin micelles,DHA carrying soybean globulin micelles and DHA carrying ag-lactoglobulin was 6 times,6.5 times,5.4 times and 6.2 times of that of DHA,respectively.Intracellular cholesterol(TC),triglyceride(TG),low-density lipoprotein cholesterol(LDL-C),alanine aminotransferase(ALT),glutamate aminotransferase(AST)and uric acid(UA)were all significantly reduced,but there was no significant effect on high-density lipoprotein cholesterol(HDL-C).Improve oil red O staining,observe intracellular lipid drops significantly reduced,cell autophagy level back to normal,the expression of autophagy marker LC3-?increases,DHA myosin micelle lipid-lowering capacity is particularly significant,followed by the DHA breast beta globulin,load DHA ovalbumin micelle and DHA glycinin micelles.In HepG2 cell oxidative stress model is established to alleviate cell oxidative stress injury were determined,and the ability of DHA ovalbumin micelle,load DHA glycinin micelle and DHA?-globulin compared micelle DHA group,the membrane potential and mitochondrial permeability transport pore(PTP)over openness,superoxide dismutase(SOD),peroxidase(CAT)and glutathione peroxidase(GSH-Px)activity were increased,pyruvic acid(PA)and malondialdehyde(MDA)content decreased significantly.At the same time,as seen under the microscope,there was a decrease in cell shrinkage and apoptosis.DHA group,DHA oboalbumin micelles,DHA myosin micelles,DHA eosin micelles and DHA eosin micelles were 14.23±1.54(×10~6U/mg prot),13.24±1.46(×10~6U/mg prot),12.41±1.43(×10~6U/mg prot),12.12±0.39(×10~6U/mg prot)and 11.18±1.21(×10~6U/mg prot),respectively,compared with the DHA group.Caspase-3 activity of DHA oval-albumin micelles,DHA myosin micelles,DHA soy globulin micelles and DHA tone-lactoglobulin micelles decreased significantly,and played a role in alleviating oxidative stress damage.Flow cytometry instrument observation and detection of cell apoptosis,finds four DHA protein micelles are reduce number of apoptotic cells and mitotic phase of G1 and G2 restoration,which carry the breast DHA beta globulin relieve oxidative stress damage the ability of the most prominent,cell stage G1 and G2 ratio increased by 8.52%and 6.11%respectively,the second is the DHA soybean globulin micelle,load DHA ovalbumin micelle and DHA glycinin micelles.Through the determination of structure,stability and biological activity,this study is of long-term significance for protecting DHA,preventing oxidative inactivation,promoting DHA to play the biological functions of reducing lipid and relieving oxidative stress.The results showed that the structure of docosahexaenoglobin was complex,the embedding amount was optimal,but the transport rate was low when it entered the cell.DHA-carrying myosin plays the best role in the process of lipid reduction with high transport rate and easy to enter cells.DHA-lactoglobulin has the best effect on relieving oxidative stress damage.The enzyme-hydrolyzed lactoglobulin can produce part of the antioxidant peptide,and the effect with DHA can improve the antioxidant,and the structure system is stable and the transport rate is fast.Egg albumin micelles have large particle size and general antioxidant property.Combining with DHA can play a certain role in lowering lipid and alleviating oxidative stress damage.