基于代谢组学研究原花青素B_(1)的肠道微生物代谢

Studies on the Intestinal Microbial Metabolism of Procyanidin B_(1) based on Metabonomics

目的:利用猪盲肠微生物通过体外静态培养的方式探究原花青素B_(1)的肠道微生物代谢。方法:在厌氧操作条件下,从新鲜屠宰的猪盲肠肠腔中获得孵育模型所需肠道微生物,配制培养基,孵育原花青素B_(1)。采集0,2,4,6,10,18 h的孵育液,用超高效液相色谱-飞行时间质谱联用仪(UHPLC-TOF-MS/MS)检测不同时间点的代谢产物,借助非靶向代谢组学分析方法,确定原花青素B_(1)代谢过程中的差异性代谢产物,并以此解析原花青素B_(1)的代谢。结果:共筛选和鉴定到21种代谢产物,其中原花青素B_(1)经肠道菌群的代谢快速,在6 h时消耗殆尽;代谢产物儿茶素和5-(3',4'-二羟基苯基)-γ-戊内酯在2~4 h间大量产生;苯丙酸和苯甲酸等为主要的代谢产物,在6 h达到峰值,在4~18 h间持续产生。通过以上结果分析原花青素B_(1)的代谢路径,原花青素B_(1)可能通过3种路径代谢,分别是:经历黄烷键断裂,下部结构形成儿茶素,儿茶素进一步发生C环开环,形成儿茶素开环产物(DHP-OL);原花青素B_(1)下部结构单元经历A环裂解,形成苯基-γ-戊内酯;原花青素B_(1)上部结构单元裂解直接形成酚酸类化合物。

The intestinal microbial metabolism of procyanidin B_(1) was explored by using porcine caecal microorganisms through static culture in vitro.Under anaerobic operating conditions,firstly,we obtained the intestinal microorganisms from the cecal lumen of freshly slaughtered pigs,which required for the incubation model,then incubated the procyanidin B_(1) in prepared the culture medium,and collected the incubation solution at 0,2,4,6,10,18 h,Use UHPLC-TOF-MS/MS to detect metabolites at different time points,and use non-targeted metabolomics analysis methods to determine differential metabolites in the metabolic process of procyanidin B_(1),and analyze metabolic pathway of procyanidin B_(1).Results showed:21 kinds of metabolites were screened and identified.The metabolism of procyanidin B_(1) by the intestinal flora was fairly rapid,and it was exhausted at 6 h;the metabolites catechins and 5-(3',4'-dihydroxyphenyl)-γ-valerolactone was produced in large quantities between 2-4 h;Phenylpropionic acid and benzoic acid were the main metabolites,which reach a peak at 6 h and continued producing between 4-18 h.Analyze the metabolic pathways of procyanidin B_(1) based on the results above.We believed that procyanidin B_(1) can be metabolized through three pathways,namely:the flavan bond breaks,the lower structure forms catechins,and the catechins further open the C ring to form catechins.C-ring-opening product;the lower structural unit of procyanidin B_(1) undergoes cleavage of the A ring to form phenyl-γ-valerolactone;the cleavage of the upper structural unit of procyanidin B_(1) directly forms phenolic acids.