目的 研究牛膽汁發(fā)酵天南星Arisaematis Rhizoma制備膽南星Arisaema cum Bile過(guò)程中微生物群落及化學(xué)成分的變化規(guī)律，為其炮制機(jī)制提供研究思路。方法 采用新鮮牛膽汁發(fā)酵炮制天南星［藥物-輔料1∶3，發(fā)酵溫度（30±5）℃，發(fā)酵濕度（70±5）%］，制備不同發(fā)酵時(shí)間的膽南星樣品。利用HPLC法對(duì)各樣品中夏佛塔苷、異夏佛塔苷、黃嘌呤、尿嘧啶等12個(gè)成分進(jìn)行含量測(cè)定，同時(shí)采用16S rRNA高通量測(cè)序技術(shù)對(duì)其中微生物組成進(jìn)行鑒定和考察。對(duì)“微生物-成分”進(jìn)行相關(guān)性分析，探討兩者在發(fā)酵炮制過(guò)程中的變化規(guī)律。結(jié)果 隨著發(fā)酵時(shí)間的增加，腺嘌呤、鳥苷、夏佛塔苷、腺苷、肌苷、胸腺嘧啶核苷、甘氨膽酸含量降低；膽酸含量增加；次黃嘌呤、黃嘌呤含量先增加后減少。菌群分析發(fā)現(xiàn)，在門水平上主要以變形菌門、厚壁菌門和藍(lán)藻菌門為主；在屬水平上主要以乳酸桿菌和假單胞菌為主。關(guān)聯(lián)分析表明，膽南星中化學(xué)成分與菌群（門、屬）間存在顯著相關(guān)，其中腺嘌呤、胸腺嘧啶核苷與藍(lán)藻菌門、放線菌門呈顯著正相關(guān)（P＜0.01），與變形菌門呈顯著負(fù)相關(guān)（P＜0.05）；膽酸與厚壁菌門呈顯著正相關(guān)（P＜0.01），與變形菌門呈顯著負(fù)相關(guān)（P＜0.05）。結(jié)論 牛膽汁發(fā)酵炮制天南星的過(guò)程中化學(xué)成分與微生物群落之間存在顯著的變化規(guī)律，為其炮制機(jī)制研究提供理論基礎(chǔ)。

Objective To study the changes of microbial community and chemical composition in the preparation of Arisaema cum Bile by fermentation of Arisaematis Rhizoma from bovine bile, and to provide research ideas for its processing mechanism. Methods Firstly, the fresh bovine bile was used to ferment and process Arisaema Rhizoma [drug-excipient ratio = 1: 3, fermentation temperature (30 ±5) ℃, fermentation humidity (70 ±5) %] to prepare Arisaema cum Bile samples with different fermentation time. Then, HPLC was used to determine the content of 12 components including schaftoside, isoschaftoside, xanthine and uracil in the samples. As well, the 16S rRNA high-throughput sequencing technology was employed to identify and to investigate the species and the abundance of microbial communities in each Arisaema cum Bile sample. Finally, the correlation analysis of “microorganism-component” was carried out via mathematical algorithm, and the changed rules of them in the processing was discussed. Results With the increase of fermentation time, the contents of adenine, guanosine, schaftoside, adenosine, inosine, thymidine and glycocholic acid decreased, the content of cholic acid increased, the content of hypoxanthine and xanthine increased first and then decreased. The results of flora analysis showed that there were mainly Proteobacteria, Firmicutes and Cyanobacteria at the phylum level. At the genus level, there were mainly Lactobacillus and Pseudomonas. Correlation analysis showed that there was a significant correlation between the chemical component of Arisaema cum Bile and the fermentation flora (phylum and genus). Adenine and thymidine were significantly positively correlated with Cyanophyta and Actinobacteria (P < 0.01), while negatively correlated with Proteobacteria (P < 0.05). Cholic acid was positively correlated with Firmicutes (P < 0.01), and negatively correlated with Proteobacteria (P < 0.05). Conclusion There were significant changes between chemical components-microbial communities during the fermentation of Arisaematis Rhizoma with bovine bile processing. The results can provide a theoretical basis for the study of the processing mechanism of Arisaema cum Bile.

R283.6

陜西省高水平中醫(yī)藥重點(diǎn)學(xué)科中藥炮制學(xué)（陜中醫(yī)藥函〔2023〕83號(hào)）;陜西省重點(diǎn)研發(fā)計(jì)劃（2024SF-ZDCYL-03-11）;秦創(chuàng)原銅川“科學(xué)家＋工程師”隊(duì)伍建設(shè)項(xiàng)目（2024KJ-KXJGCS-01）;王昌利全國(guó)老藥工傳承工作室（國(guó)中醫(yī)藥人教函〔2024〕255號(hào)）;秦創(chuàng)原中醫(yī)藥產(chǎn)業(yè)創(chuàng)新聚集區(qū)項(xiàng)目（L2024-QCY-ZYYJJQ-X34）