Li Zhe, Wang Yuqing, Wang Zicheng, Wu Dongxue, Zhao Yuhao, Gong Xun, Jiang Quan, Xia Congmin
Guang'an Men Hospital, China Academy of Chinese Medical Sciences, Beijing, China.
School of Pharmacy, Shandong University of Traditional Chinese Medicine, Jinan, China.
Front Pharmacol. 2024 Oct 9;15:1437020. doi: 10.3389/fphar.2024.1437020. eCollection 2024.
Genistin, as a kind of natural isoflavone glycoside, has good biological activity, and its weak absorption makes it closely related to intestinal flora. However, the role of the intestinal flora is still unclear and whether the metabolites produced by the intestinal flora are absorbed systemically is also variable.
Genistin was fermented for 24 h based on fecal bacteria fermentation technology. The components were qualitatively and quantitatively analyzed by HPLC and UHPLC-Q-Exactive Orbitrap Mass spectrometry. The composition of intestinal flora in fermentation samples from fecal bacteria was detected by 16S rRNA sequencing. Five representative probiotics were cultured and fermented with genistin to determine similarities and differences in genistin metabolites by different bacteria at different times. Finally, the absorption results of metabolites by fermentation were verified by a Caco-2 cell monolayer.
The HPLC results of fecal fermentation showed that genistein levels increased from 0.0139 ± 0.0057 mg/mL to 0.0426 ± 0.0251 mg/mL and two new metabolites were produced. A total of 46 metabolites following fecal fermentation were identified, resulting from various biotransformation reaction products, such as decarbonylation, hydroxylation, and methylation. Simultaneously, the 16S rRNA results showed that the intestinal flora changed significantly before and after fermentation and that the intestinal microorganisms in the control (Con) group and the fermentation (Fer) group showed a significant separation trend. Five genera, , , , , and , were considered the dominant flora for genistin fermentation. The qualitative results of fermentation of genistin by five probiotics at different times showed that there were significant differences in small molecule metabolites by fermentation of different bacteria. Meanwhile, most metabolites could be identified following fecal bacteria fermentation, which verified the importance of the dominant bacteria in the feces for the biotransformation of components. Finally, the absorption results of the metabolites based on the Caco-2 cell monolayer showed that 14 metabolites could be absorbed into the circulation in vivo through the mesentery.
The small molecule metabolites of genistin by fermentation of fecal bacteria can be well absorbed systemically by the body. These studies provide a reference value for explaining the transformation and absorption of flavonoid glycosides in the intestine.
染料木苷作为一种天然异黄酮糖苷，具有良好的生物活性，其吸收较弱使其与肠道菌群密切相关。然而，肠道菌群的作用仍不明确，且肠道菌群产生的代谢产物是否被全身吸收也存在差异。
基于粪便细菌发酵技术对染料木苷进行24小时发酵。通过高效液相色谱法（HPLC）和超高效液相色谱-四极杆-静电场轨道阱质谱联用仪（UHPLC-Q-Exactive Orbitrap Mass spectrometry）对成分进行定性和定量分析。通过16S rRNA测序检测粪便细菌发酵样品中的肠道菌群组成。培养5种代表性益生菌并用染料木苷进行发酵，以确定不同细菌在不同时间对染料木苷代谢产物的异同。最后，通过Caco-2细胞单层验证发酵代谢产物的吸收结果。
粪便发酵的HPLC结果显示，染料木素水平从0.0139±0.0057mg/mL增加到0.0426±0.0251mg/mL，并产生了两种新的代谢产物。粪便发酵后共鉴定出46种代谢产物，这些代谢产物是由脱羰基、羟基化和甲基化等各种生物转化反应产物产生的。同时，16S rRNA结果显示，发酵前后肠道菌群发生了显著变化，对照组（Con）和发酵组（Fer）的肠道微生物呈现出明显的分离趋势。五个属，即[此处原文缺失五个属的具体名称]，被认为是染料木苷发酵的优势菌群。5种益生菌在不同时间对染料木苷发酵的定性结果表明，不同细菌发酵产生的小分子代谢产物存在显著差异。同时，大多数代谢产物可在粪便细菌发酵后被鉴定出来，这证实了粪便中优势菌对成分生物转化的重要性。最后，基于Caco-2细胞单层的代谢产物吸收结果表明，14种代谢产物可通过肠系膜被吸收进入体内循环。
粪便细菌发酵产生的染料木苷小分子代谢产物可被机体很好地全身吸收。这些研究为解释黄酮类糖苷在肠道中的转化和吸收提供了参考价值。
