北苍术米泔水炮制前后化学成分变化及其对脾虚泄泻大鼠肠道真菌菌群的影响

Analysis of chemical compositions in Atractylodes chinensis with rice water before and after processing and its effects on intestinal fungal flora of spleen deficiency diarrhea rats

目的 探究北苍术Atractylodes chinensis米泔水炮制前后成分变化及其对大黄所致脾虚泄泻模型大鼠肠道真菌菌群的影响。方法 采用气相色谱串联四极杆质谱仪联用技术(GC-MS)和HPLC法对北苍术生品及其米泔水炮制品化学成分进行对比分析,利用ITS高通量测序技术,探究其对大黄所致脾虚泄泻模型大鼠肠道真菌群落变化的影响。结果 GC-MS和HPLC结果显示,炮制前后化学成分类型没有改变,而成分含量多有变化,其中挥发油类(茅术醇、β-桉叶醇、α-红药没醇、苍术酮)炮制后含量显著降低(P<0.05),多数酯类成分炮制后含量增加,可能与北苍术“炮制减燥”有关。高通量测序结果显示,属水平上,生北苍术组真菌群落结构与模型组相似,而米泔水制北苍术组更接近于空白组,说明米泔水制北苍术能够调节大黄所致的脾虚型腹泻,对于肠道真菌菌群的调控能力优于生北苍术。Pearson相关性分析表明,部分真菌菌群与活性成分含量相关,如线黑粉酵母属Filobasidium sp.、链格孢属Alternaria sp.真菌与米泔水制北苍术中白术内酯II含量显著正相关(P<0.05),与苍术酮含量呈显著负相关(P<0.05)。结论 北苍术米泔水炮制前后化学成分含量存在差异,其中茅术醇、β-桉叶醇、苍术酮与北苍术炮制减燥及治疗脾虚泄泻疾病有密切关系,米泔水制北苍术对模型大鼠肠道真菌调控能力优于生北苍术,该研究结果可为扩大米泔水制苍术临床应用提供理论参考。

Objective To explore the changes of composition in Atractylodes chinensis(AC) with rice water before and after processing and its effect on intestinal fungal flora of rats with spleen deficiency and diarrhea caused by Dahuang(Rhei Radix et Rhizoma,RRR).Methods The chemical components of the raw products and processed products of AC were analyzed by GC-MS and HPLC technologies.ITS high-throughput sequencing technology was used to explore the effects of them on intestinal fungal community changes in rats with spleen deficiency diarrhea induced by RRR.Results The results of GC-MS and HPLC showed that there was no change in the type of chemical components before and after processing,but the content of chemical components changed.The volatile contents of hinesol,β-eudesmol,α-bisabolol and atractylone decreased significantly(P < 0.05) and the content of most esters increased after processing.Maybe it related to “processing reduced the dryness”.At the genus level,high-throughput sequencing results showed that the fungal community structure of AC group was similar to the model group,and the AC prepared by rice water was closer to the blank group,indicating that AC processed by rice water could regulate spleen deficiency diarrhea caused by RRR,and the regulation ability of intestinal fungal flora was better than the raw products.Pearson correlation analysis showed that some fungal groups were correlated with the content of active ingredients.There was a significantly positive correlation between Filobasidium sp.,Alternaria sp.and atractylenolide II content,the correlation with atractylone content was significantly negative.Conclusion There were differences in chemical composition content of AC with rice water before and after processing.Among them,hinesol,β-eudesmol and atractylone were closely related to AC desiccation and treating spleen deficiency and diarrhea diseases.And the regulation ability of intestinal fungi in the model rats with AC processed by rice water was better than that raw products of AC.The results can provide theoretical reference for expanding the clinical application of rice water processed AC.