Protein Composition Analysis And Functions Of Four Bee Venoms

Among the many bee products,bee venom is one of the most valuable products for development and utilization,and it is also a most important biotoxin.However,due to the intricate composition of bee venom,including a variety of enzymes,peptides,amines and other substances,the development and utilization of bee venom are limited.Therefore,this paper studied the protein components of four bee venoms,and evaluated the efficacy of four bee venoms in anti-aging and anti-inflammatory,providing theoretical basis for the development and utilization of bee venom in the future.The main conclusions of this paper include the following five parts:(1)In this paper,the protein components in the four bee venoms were analyzed by Native-PAGE electrophoresis.The results show,two kinds of proteins were isolated from Vespa mandarinia Smith venom,three kinds of proteins were isolated from Vespa bicolor Fabricius venom,three kinds of proteins were isolated from Xylocopa(Nyctomelitta)tranquebarica venom,and one protein was isolated from Apis mellifera venom.Meanwhile,five kinds of proteins were isolated from Vespa mandarinia Smith venom,five kinds of proteins were isolated from Vespa bicolor Fabricius venom,five kinds of proteins were isolated from Xylocopa(Nyctomelitta)tranquebarica venom,and four proteins were isolated from Apis mellifera venom by SDS-PAGE electrophoresis.In addition,the separation effects of three pre-formed gels on bee venom were compared.The results showed that the separation of Tris-glycine pre-formed gel was the best.(2)The native-PAGE and SDS-PAGE protein bands of four bee venoms were identified by high performance liquid chromatography-mass spectrometry(LC-MS).Among them,the five protein components of the three species of Vespidae venom are the same,which are dipeptidyl peptidase ?,hyaluronidase,phosphatase A,allergen antigen 5,and cytochrome c oxidase subunit 1.However,there are differences in homology between different bee venom components.Five protein components were also identified in Apis mellifera venom,namely venom dipeptidyl peptidase 4,hyaluronidase,phosphatase Ai,phospholipase A2,and melittin.It differs from the three species of Vespidae venom in protein composition.(3)The enzyme activities of hyaluronidase and phosphatase A in four bee venoms were detected by enzyme-linked immunosorbent assay.The results showed that the hyaluronidase and phosphatase A activities of Vespa bicolor Fabricius venom were the highest,300.41 ±7.03 U/g and 8.89±0.51 U/g respectively.The enzyme activity of the hyaluronidase and phosphatase A of Apis mellifera venom was the lowest,8.89±0.51 U/g and 2.5810.16 U/g,respectively.(4)In vitro experiments were performed using human dermal fibroblasts(HDF)to evaluate the anti-aging effects of the four bee venoms.The effects of four bee venoms on the secretion of type I collagen(Col-I)and matrix metalloproteinase(MMP-1)by HDF cells were examined by enzyme-linked immunosorbent assay kit.The results showed that among the four bee venoms,only the anti-aging of Vespa mandarinia Smith and Vespa bicolor Fabricius venom have obvious effects,while Xylocopa(Nyctomelitta)tranquebarica and Apis mellifera venom have no significant effect.When the concentration of Vespa mandarinia Smith and Vespa bicolor Fabricius venom was 10 mg/L,the secretion of Col-I by HDF cells increased by 24.62%and 30.31%,respectively.After treatment with 10 mg/L Vespa mandarinia Smith and Vespa bicolor Fabricius venom,the content of MMP-1 secreted by the cells was 937.29 p.g/L and 784.15 ?g/L,respectively,which was reduced by 26.03%and 43.64%,respectively,compared with the blank group.(5)A cellular inflammation method of inducing human umbilical vein endothelial cells(HUVEC)by lipopolysaccharide(LPS)was established to evaluate the anti-inflammatory effects of four bee venoms.The effects of four bee venoms on the inflammatory factors nitric oxide(NO),human interleukin-6(IL-6)and human interleukin-8(IL-8)secreted by the cells were examined by enzyme-linked immunosorbent assay kit.The results showed that the four bee venoms could reduce the content of inflammatory cytokines secreted by HUVEC cells.When the concentration of Apis mellifera venom was 5 mg/L,the content of IL-6 decreased by 62%compared with the model group.When the concentration of Apis mellifera venom was 10 mg/L,the effect of inhibiting IL-8 secretion was the most obvious,which reduced the IL-8 content of the model group by 68%.