Extraction And Purification Of Acanthopanax Senticosus Fructose Polysaccharide And Its In Vitro Anti-lung Cancer Activity

In this project,we hope to obtain a kind of polysaccharide with high activity in the fruit part of Acanthopanax senticosus and analyze its molecular weight,functional group composition and monosaccharide composition.We reported the anti-lung cancer activity of Acanthopanax senticosus fructose for the purpose of the A549 cells,and its anti-lung cancer mechanism was studied from the three aspects.For the purpose of anti-lung cancer activity,the crude polysaccharides obtained by three different extraction methods influenced on A549 cells.The polysaccharide which extrated by hot water had the highest inhibitory rate on A549 cells.And then optimize the extraction conditions of hot water extraction method.Acanthopanax senticosus fruit was extracted under optimized conditions to obtain crude polysaccharides.We used the Sevage method to defecate the protein,column chromatography methods was used to Purified polysaccharide.Binding activity tracking obtained a homogeneous molecular weight polysaccharide ASPF,and the sugar content in ASPF was 89.8%,and the uronic acid content was 22.9%.Gel permeation chromatography was used to identification of purity of ASPF.The chromatogram showed only one peak and the peak shape was great,indicating that ASPF had a uniform molecular weight,and used GPC to know the molecular weight of ASPF was 97462.The results show that the glycosyl in ASPF is in the pyran configuration,and the end group anomeric carbon is in theβ-type.Six monosaccharides in ASPF was about:Rha,Gal UA,Glc,Xyl,Gal,and Ara,with a molar ratio of 1:3.9:1.5:4.45:0.56:3.62.It is speculated that ASPF is a pectin polysaccharide.Finally,the CCK-8 method preliminarily proves the inhibitory effect of ASPF on A549 cells.The number of A549 cells gradually decreases with more concentration and time of administration.It is significant at 25μg/mL and extremely significant at200μg/mL.Its IC₅₀ value is 424.39μg/mL,compared with 664.6μg/mL before purification,its activity is significantly improved.In addition,ASPF can also inhibit the migration rate of A549 cells.Caspase enzyme activity and the m RNA expression of apoptosis genes are in the detection,it is found that ASPF may increase the m RNA expression of Bax and make the m RNA expression less on Bcl-2 to significantly enhance the activity of Caspase-3,8,and 9 enzymes to achieve Jointly promote the effect of cell apoptosis.This paper analyzed the structure of ASPF,a water-soluble polysaccharide extracted from Acanthopanax senticosus fruit,and reported its activity and mechanism in inhibiting lung cancer A549 cells in vitro,proving the medicinal value and development potential of Acanthopanax senticosus fruit.