千金子生品和霜品提取物中主要二萜醇酯类成分在大鼠肠道菌群中的体外代谢研究

Study on in vitro metabolism of main diterpenoid alcohol ester composition from Euphorbiae Semen and Euphorbiae Semen pulveratum extracts in intestinal flora of rats

目的:通过大鼠肠道菌群体外实验,分析千金子生品和霜品提取物中主要二萜醇酯类成分在大鼠肠道菌群中的代谢情况。方法:建立同时测定千金子素L_(1)、L_(2)、L_(3)、L_(7a)、L_(7b)、L_(8)的HPLC定量分析方法,并用该方法测定千金子生品和霜品提取物中各成分在体外大鼠肠道菌群作用下各个时间点的含量,随后基于UHPLC-Q-Exactive HF,进一步分析千金子制霜前后主要二萜醇酯类成分的体外代谢情况。结果:千金子生品与霜品提取物中千金子素L_(1)、L_(2)、L_(3)、L_(7a)、L_(7b)、L_(8)在体外的代谢产物一致,但代谢速率不同,千金子霜品组在代谢初期代谢速率最大。采用Q-Exactive HF结合Xcalibur软件共分析鉴定出了6个代谢产物,千金子制霜前后提取物中6个二萜醇酯类成分在体外的主要代谢途径为水解和甲基化。结论:千金子生品与霜品提取物能够在大鼠肠道菌群作用下转化为多种代谢产物,该结果为进一步研究其在体内的作用机制提供实验依据。

Objective:The metabolism of main diterpenoid alcohol ester composition from Euphorbiae Semen and Euphorbiae Semen pulveratum extracts was analyzed by in vitro experiment of intestinal flora in rats.Methods:To establish a HPLC quantitative analysis method for simultaneous determination of Euphorbia factor L_(1),L_(2),L_(3),L_(7a),L_(7b)and L_(8),and determined the contents of each component in Euphorbiae Semen and Euphorbiae Semen pulveratum extracts at various time points in vitro under the action of intestinal flora of rats.The in vitro metabolism of diterpenoid alcohol esters was further analyzed by UHPLC-Q-Exactive HF.Results:The metabolites of Euphorbia factor L_(1),L_(2),L_(3),L_(7a),L_(7b)and L_(8)of Euphorbiae Semen and Euphorbiae Semen pulveratum extracts were the same in vitro,but the metabolic rates were different.The metabolic rates of Euphorbiae Semen pulveratum extract group were the highest in the initial stage of metabolism.Six metabolites were identified by Q-Exactive HF combined with Xcalibur software,and the main metabolic pathways of the six diterpenoid alcohol esters from Euphorbiae Semen and Euphorbiae Semen pulveratum were hydrolysis and methylation in vitro.Conclusion:The Euphorbiae Semen and Euphorbiae Semen pulveratum extracts could be transformed into various metabolites under the action of intestinal microflora of rats.These results will provide reference for further study of its mechanism in vivo.