基于普雷沃氏菌评价粉葛多糖益生元活性

Evaluation of the prebiotic efficacy of Radix Puerariae thomsonii polysaccharide based on Prevotella

目的:考察粉葛多糖对普雷沃氏菌的体内外促生长作用,从而评价粉葛多糖益生元活性。方法:收集普通喂养小鼠、高脂加乙醇喂养小鼠、粉葛多糖干预高脂加乙醇喂养小鼠8周后的盲肠内容物进行16S rRNA肠道菌群测序分析。选取两株普雷沃氏菌Prevotella copri和Prevotella oralis,配制含粉葛多糖0.2%、0.4%的BHI培养液(0.2%BHI+RPP和0.4%BHI+RPP),配制不含粉葛多糖的BHI培养液(BHI)作为空白对照,对二株普雷沃氏菌进行体外培养。分别于0、3、6、9、12、24、48 h,取样测定OD值(600 nm)和pH值,另将24、48 h的样品留样,用于琥珀酸、乙酸的测定。结果:体内实验表明,粉葛多糖的摄入显著提高了小鼠肠道普雷沃氏菌的丰度。体外实验表明,BHI+0.2%RPP、BHI+0.4%RPP两组比BHI组OD值更高,pH值更低,说明粉葛多糖能促进两株普雷沃氏菌的生长,且产生了更多的酸性代谢产物。此外,BHI+0.2%RPP、BHI+0.4%RPP两组24 h和48 h的培养液中乙酸和琥珀酸的含量均比BHI组的含量高,且大部分具有显著性差异。结论:粉葛多糖体内外对普雷沃氏菌均有促生长作用,推测其具有潜在的益生元活性,为中药多糖作为益生元的研发提供理论基础。

Objective:To investigate the in vivo and in vitro growth-promoting effects of Radix Puerariae thomsonii polysaccharide on Prevotella,and to evaluate the prebiotic activity of Radix Puerariae thomsonii polysaccharide.Methods:The cecal contents of ordinary-fed mice,high-fat and alcohol-fed mice,and high-fat and alcohol-fed mice with Radix Puerariae thomsonii polysaccharide intervention were collected for 16S rRNA intestinal flora sequences after 8 weeks.Select Prevotella.copri and Prevotella.oralis and formulate BHI culture media containing 0.2%and 0.4%Radix Puerariae thomsonii polysaccharide(0.2%BHI+RPP and 0.4%BHI+RPP),and formulate BHI culture media(BHI)without Radix Puerariae thomsonii polysaccharide as control group.Two strains of Prevotella were cultured in vitro.OD(600 nm)and pH were measured at 0,3,6,9,12,24,48 h,respectively.In addition,samples of 24 h and 48 h were reserved for the determination of succinic acid and acetic acid.Results:In vivo experiments showed that the intake of Radix Puerariae thomsonii polysaccharides significantly increased the abundance of Prevotella in the intestines of mice.In vitro experiments showed that the BHI+0.2%RPP and BHI+0.4%RPP groups with Radix Puerariae thomsonii polysaccharide added had higher OD and lower pH than the control group,indicating that Radix Puerariae thomsonii polysaccharide could promote the growth of two strains of Prevotella and produce more acidic metabolites.What’s more,the contents of acetic acid and succinic acid in the culture medium of the 24 h and 48 h groups of BHI+0.2%RPP and BHI+0.4%RPP with Radix Puerariae thomsonii polysaccharides were higher than those in the control group,and most of them had significant differences.Conclusion:Radix Puerariae thomsonii polysaccharide has a growth-promoting effect on Prevotella both in vivo and in vitro,and it is speculated that it has potential prebiotic activity.It will provide a theoretical basis for the future research and development of traditional Chinese medicine polysaccharides as prebiotics.