| Bee venom active peptides are the main substance of bee venom,have many functions,such as anti-inflammatory,anti-cancer and anti-radiation.Propolis is rich in flavonoids,have many functions,such as antibacterial,antioxidant,anti-cancer.In this paper,the separation,purification and extraction were used to get venom active peptides and propolis flavonoids respectively by using bee venom and propolis as raw materials,this paper was taking the functions of antioxidant anti-gout and antirheumatic as determining target.Get the compound by mixing bee venom active peptides and propolis flavonoids together.Main results of this study are as follows:1.In this experiment,the bee venom was isolated and purified by Sephadex gel,alcohol deposition and RT-HPLC.The optimal separation method is as follows:Sephadex G-25 gel filtration was selected,the loading volume and elution rate is 28 m L/h,the sampling amount is 1.0 m L,the loading concentration is 1.0 mg/mL,the bee venom can be divided into two components P1,P2.The P1 was collected by freeze-drying with 3 times anhydrous ethanol,centrifuged at 3000 r/min for 5 min,and the precipitate was the melittin.The relative molecular weight of the melittin was about 2840 Da and the extraction rate was 96%,and the purity was 98%.The supernatant was further purified by RT-HPLC,column: Zorbax SB-C18(4.6 x 250 ?m),injection volume: 10 ?L,acetonitrile-0.1% trifluoroacetic acid as mobile phase,The three components Q1,Q2,Q3.MCD peptide,bee venom peptide and sedative peptide were obtained.The relative molecular weights were 2592 Da,2035 Da and about 2700 Da,respectively.The extraction rates were 78.6%,89%,70%.The purity of MCD peptide and bee venom was 91.28 % and 90.16%,respectively.2.In this experiment,the best extraction conditions of propolis flavonoids by studying on single factor and response surface were as follows: ethanol concentration 82%,extraction time 3 h,extraction temperature 83?,ratio of solid to liquid 1:10,in this condition,The extraction rate of propolis flavonoids was 46.30%.3.The studying of bee venom active peptide components of antioxidant,anti-gout,anti-rheumatoid arthritis physiological activity.The results showed that the clearance rate of hydroxyl radical scavenging was 0.32 mg/mL and the scavenging rate of superoxide anion was 16.04%.The same concentration of melycotin and MCD peptide on hydroxyl radical scavenging rate 3.51%,1.14%,the superoxide anion clearance rate was 4.51%,1.34%.Melittin is a major antioxidant substance in bee venom.Bee venom peptide,melittin peptide,MCD peptide high dose group compared with the model group,the activity of XOD decreased by 46.94%,42.00% and 31.54%,and the content of UA decreased by 62.01%,57.38% and 59.75%,respectively,compared with the model group.The content of CRE was 14.63%,19.34% and 20.47% higher than that of the blank group,and the content of BUN was 12.79%,19.16% and 17.17% higher than that of the blank group.The main substance of anti-gout in bee venom active peptide is melittin by this experiment.The levels of TNF-? were decreased by 28.32%,24.68% and 23.29%,and the content of IL-6 was decreased by 54.52% compared with the model group in the high dose group of melittin,bee venom peptide and MCD peptide.51.97%,50.99%,IL-1? decreased by 26.60%,20.81% and 18.94% respectively.Indicating that the main substance of anti-rheumatoid arthritis in bee venom is melittin.4.Ropolis flavonoids antioxidant,anti-gout,anti-rheumatoid arthritis physiological activity research.The results showed that 0.83 mg/m L propolis flavonoids had a scavenging rate of 96.39% for hydroxyl radicals and 91.89% for superoxide anion.Indicating that propolis flavonoids have antioxidant effects.The experimental results showed that the XOD activity of propolis flavonoids was 45.58% lower than that of the model group and the UA content was 34.38% lower than that in the blank group.The CRE content increased by 7.69% and the BUN content increased by 17.91%.So propolis flavonoids have a role in fighting gout.The results showed that the content of TNF-? in the high dose of propolis flavonoids was 28.08% lower than that in the model group,the content of IL-6 was decreased by 49.44% and the content of IL-1? decreased by 18.45%.Indicating that propolis flavonoids have the effect of fighting rheumatoid arthritis.5.Bee venom propolis antioxidant,anti-gout,anti-rheumatoid arthritis physiological activity complex conditions.Antioxidant experiments showed that 0.2 mg/m L bee venom and 4.15 mg/mL propolis flavonoids were arranged at 1:5,the removal rate of · OH was 96.40%,the superoxide anion free radical scavenging rate was 91.90%.Mice gout experiments showed that 0.05 mg/m L bee venom and 0.326 mg/m L propolis flavonoids were formulated at a ratio of 1:8.Compared with the model group,the XOD activity decreased by 56.75% and the UA content decreased by 46.81% in the 1: 8 group.The contents of CRE and BUN increased by 8.66% and 25.78% respectively compared with the blank group.Indicating that the compound product has the effect of relieving gout,and does not cause kidney damage.Mice rheumatoid arthritis experiments showed that 0.05 mg/mL bee venom and 0.326 mg/m L propolis flavonoids were formulated at a ratio of 1:7.Compared with the model group,the levels of serum TNF-?,IL-6 and IL-1? decreased by 33.28%,50.60% and 15.81%,respectively in the 1:7 group.Indicating that the compound product has the effect of relieving rheumatoid arthritis.At present,the risk of chronic diseases such as gout and rheumatoid arthritis is increasing year by year.It is important to study the compound products of relieving gout and rheumatoid arthritis.This is the theoretical basis on the development and utilization of bee venom and propolis products. |
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