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UHPLC-LTQ-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 ketone 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-atractylenolide Ⅰ, 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-atractylenolide Ⅰ, 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, and there were differences in the metabolic transformation degree of two components, and most components had a higher transformation (or formation) rate in ethanol extract of deep-fried Atractylodis Rhizoma, which may be the key reason for the enhancement of intestinal strengthening and anti-diarrhea effect of ethanol extract of deep-fried Atractylodis Rhizoma. The metabolic product identification results of atractyloside A further verified the key role of intestinal flora in the influence of components.

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）