体外共孵育法研究肠道菌群对苍术炒焦前后醇提物及增量成分苍术苷A的代谢规律  

Metabolic regularity of intestinal flora on alcohol extracts of Atractylodis Rhizoma before and after stir-frying and incremental component atractyloside A by in vitro co-incubation method

作　　者：曹皇亮 刘春莲 陈高源 姚丁 肖扬鑫 刘艳菊 CAO Huangliang;LIU Chunlian;CHEN Gaoyuan;YAO Ding;XIAO Yangxin;LIU Yanju(College of Pharmacy,Hubei University of Chinese Medicine,Wuhan 430065,China;Hubei Engineering Research Center of Chinese Material Medica Processing Technology,Wuhan 430065,China;Hubei Shizhen Laboratory,Wuhan 430065,China)

机构地区：[1]湖北中医药大学药学院,湖北武汉430065 [2]湖北省中药炮制技术工程中心,湖北武汉430065 [3]湖北时珍实验室,湖北武汉430065

出　　处：《中草药》2024年第8期2641-2650,共10页Chinese Traditional and Herbal Drugs

基　　金：国家自然科学基金资助项目(82074018);湖北省中央引导地方发展项目(2020ZYYD030)。

摘　　要：目的明确生、焦苍术醇提物在肠道菌群作用下成分的代谢差异,以及关键增量成分苍术苷A的代谢转化规律。方法采用体外共孵育方法,将生苍术、焦苍术醇提物以及炒焦增量成分苍术苷A与小鼠肠道菌液共孵育,运用UHPLCLTQ-Qrbitrap技术,分析生苍术、焦苍术醇提物共孵育后的转化情况,并对苍术苷A的代谢产物进行鉴定分析。结果生苍术、焦苍术醇提物与小鼠肠道菌群共孵育后原型成分苍术苷A、5-羟甲基糠醛、苍术素、白术内酰胺、2-(联苯基-4-基)乙醛、二乙酰苍术素醇、白术内酯II、羟甲香豆素、乙酰苍术素醇、白术内酯I、β-石竹烯、3β-乙酰氧基苍术酮、苍术酮13种成分峰面积均降低,其中,苍术素、二乙酰苍术素醇、羟甲香豆素、苍术酮4个成分在焦苍术中降低尤为明显;3β-乙酰氧基-白术内酯I、β-香根草烯、白术内酯III、愈创木烯、芹烷二烯酮5个成分峰面积在生、焦苍术醇提物与菌群共孵育后均增加,其中,3β-乙酰氧基-白术内酯I、愈创木烯、芹烷二烯酮在焦苍术中增加尤为明显;苍术苷A经肠道菌群代谢转化产生了新成分,根据数据分析,推测出其中与其羟化、羰基化、乙酰化相关的4种代谢物。结论肠道菌群对生苍术、焦苍术醇提物成分有转化作用,且对2种饮片成分代谢转化程度存在差异,大多成分在焦苍术中转化(或生成)率更高,这可能是焦苍术增强固肠止泻作用关键原因;苍术苷A的代谢产物鉴定结果进一步验证了肠道菌群对成分影响的关键作用。Objective To clarify the metabolic differences of ethanol extract of Cangzhu(Atractylodis Rhizoma)and ethanol extract of deep-fried Atractylodis Rhizoma under the action of intestinal flora,as well as the metabolic transformation rules of the key component atractyloside A.Methods This study adopted further in vitro co-incubation method to co-incubate ethanol extract of Atractylodis Rhizoma,ethanol extract of deep-fried Atractylodis Rhizoma,and atractylodes A,the incremental component of stir-coke,with mouse intestinal bacterial solution.UHPLC-LTQ-Orbitrap technology was used to analyze the transformation of ethanol extract of Atractylodis Rhizoma and ethanol extract of deep-fried Atractylodis Rhizoma after co-incubation and the metabolites of atractyloside A were identified and analyzed.Results The peak area of 13 components of atractyloside A,5-hydroxymethylfurfural,atractylodin,atractylenolactam,2-(bipheny-4-yl)acetaldehyde,diacetylatractylodinol,atractylenolide II,hymecromone,acetylatractylodinol,atractylenolide I,β-caryophyllene,3β-acetoxyatractyloketone,atractylodes keton e were all decreased after co-breeding with the intestinal flora of mice.Atractylodin,diacetylatractyloside alcohol,hymecromone,atractylon were particularly decreased in deep-fried Atractylodis Rhizoma.The peak area of 3β-acetoxy-atractylenolideⅠ,β-vetiverene,atractylenolide III,guaiene and 7(11)-dien-8-one increased after co-culture of ethanol extract of Atractylodis Rhizoma and ethanol extract of deep-fried Atractylodis Rhizoma with bacterial community,and the increase of 3β-acetoxy-atractylenolideⅠ,guaiene and selina-4(14),7(11)-dien-8-one was particularly obvious in deep-fried Atractylodis Rhizoma.Atractyloside A was metabolized by intestinal flora to produce new components.According to the data analysis,four metabolites related to hydroxylation,carbonylation and acetylation were deduced.Conclusion The intestinal flora had a transformation effect on ethanol extracts of Atractylodis Rhizoma and deep-fried Atractylodis Rhizoma,an

关 键 词：苍术 焦苍术 苍术苷A 肠道菌群 成分转化 体外共孵育法 UHPLC-LTQ-Qrbitrap技术 苍术素 二乙酰苍术素醇 羟甲香豆素 苍术酮 3β-乙酰氧基-白术内酯Ⅰ β-香根草烯 白术内酯Ⅲ 愈创木烯 芹烷二烯酮 

分 类 号：R283.6[医药卫生—中药学]