银杏双黄酮和银杏内酯B与人体肠道菌群体外互作研究

Study the interactions between ginkgetin and ginkgolide B with human gut microbiota in vitro

摘要:

[目的] 银杏提取物在防治心血管系统和神经系统疾病方面发挥重要功能。鉴于肠道菌群已被认定为一个新兴的药物作用靶标，研究银杏双黄酮和银杏内酯与人体肠道菌群之间的相互作用具有非常重要的意义，这将为进一步理解银杏提取物的功能和作用机制奠定基础。[方法] 本研究使用人体肠道菌群体外批量发酵、细菌总量测定、细菌16S rDNA高通量测序、气相色谱和液相色谱检测等方法，对银杏双黄酮和银杏内酯B单独或复合在体外与人体肠道菌群的相互作用进行研究。[结果] 银杏双黄酮和银杏内酯B单独添加对人体肠道菌群总量、肠道菌群结构组成和短链脂肪酸产量没有显著影响。但有意思的是，复合添加银杏双黄酮和银杏内酯B后，Coriobacteriaceae科和Cupriavidus属细菌的比例显著升高，Gemella菌细菌比例显著降低。功能基因预测分析发现，编码K00076、K12143、K07716和K00220的基因在复合添加银杏双黄酮和银杏内酯B后显著富集。K00076和K00220是氧化还原酶，催化CH-OH供体基团的电子转移，可能参与银杏双黄酮和银杏内酯B的代谢和修饰。HPLC检测发现，人体肠道菌群体外对银杏双黄酮和银杏内脂B的降解修饰率分别为70%和35%左右。[结论] 体外复合添加银杏双黄酮和银杏内酯B可显著改变肠道某些细菌的丰度。同时，体外研究表明肠道菌群具有代谢修饰银杏双黄酮和银杏内酯B的功能。

关键词:银杏双黄酮;银杏内酯B;人体肠道菌群

Abstract:

[Objective] Ginkgo biloba extract plays an important role in the prevention and treatment of cardiovascular and nervous system diseases. In consideration of the fact that gut microbiota has been identified as a new drug target, it is of great significance to study the interactions between ginkgetin and ginkgolide B with human gut microbiota, which will lay a foundation for further understanding the function and mechanism of Ginkgo biloba extract. [Methods] In this study, batch fermentation, bacterial amount detection, 16S rDNA high-throughput sequencing, GC and HPLC measurement were used to study the interactions between ginkgetin and/or ginkgolide B with human gut microbiota in vitro. [Results] Neither ginkgetin nor ginkgolide B alone had a significant effect on the total amount of gut microbiota, the composition of intestinal flora and the production of short chain fatty acids by the gut microbiota. However, when ginkgetin and ginkgolide B were added in combination, the proportion of bacterial represented by the family Coriobacteriaeae and genus Cupriavidus increased significantly, while the proportion represented by the genus Gemella decreased significantly. Functional gene prediction analysis found that genes encoding K00076, K12143, K07716, and K00220 significantly enriched in the presence of ginkgetin and ginkgolide B. Moreover, K00076 and K00220 are oxidoreductases that catalyze the transfer of electrons from the CH-OH donor group and may be involved in the metabolism and modification of ginkgetin and ginkgolide B. The degradation and modification rates of ginkgetin and ginkgolide B by human gut microbiota in vitro were ~70% and ~35%, respectively. [Conclusion] The relative percentage of some intestinal bacteria was significantly changed by the combined addition of ginkgetin and ginkgolide B in vitro. Meanwhile, human gut microbiota has the function on metabolism or modifying ginkgetin and ginkgolide B.

Key words:ginkgetin;ginkgolide B;human gut microbiota