不同培养基条件下三七根总皂苷与人肠道菌群的体外相互作用

Interaction between Panax notoginseng Root Saponins and Human Gut Microbiota under Different Culture Conditions in Vitro

摘要: 三七根总皂苷(Panax notoginseng root saponins, PNS)作为三七的主要活性成分，口服后会与肠道菌群发生相互作用，但目前关于不同营养条件下二者互作的认识尚不充分。本研究采用BHI、GAM及GMM培养基对人源粪便菌群进行体外厌氧培养，并分别设立添加PNS的实验组(BHIP, GAMP, GMMP)。通过高效液相色谱定量分析不同培养基条件下5种高含量皂苷的相对代谢率，并用16S rRNA测序解析菌群结构变化。结果显示：BHIP组对皂苷R₁、Rb₁和Rd的平均相对代谢率 > 73%；GAMP组特异性高效代谢Rb₁，代谢率 > 84%；而GMMP组仅代谢R₁和Re，且代谢率 < 25%。BHI、GAM与GMM三组所培养的菌群结构存在显著差异；与各自对照组相比，BHIP与GAMP组的菌群α多样性(Shannon指数)显著降低(P < 0.01)，群落结构发生显著重构(P < 0.001)，而GMMP组则无显著变化。在菌群组成上，BHIP组显著富集了Lachnoclostridium、Parasutterella和Bifidobacterium；GAMP组显著富集了Bifidobacterium、Escherichia-Shigella；GMMP组则主要富集了Roseburia和Anaerostipes。综上，PNS与肠道菌群的互作效应在不同培养条件下呈现出显著的差异性，在评估PNS的益生效应时，需充分考虑其环境背景。

Abstract: Panax notoginseng root saponins (PNS), the main active components of P. notoginseng, interact with the gut microbiota after oral administration. However, the understanding of their interaction under different nutritional conditions remains limited. In this study, human fecal microbiota was anaerobically cultured in vitro using BHI, GAM, and GMM media, with PNS added to the experimental groups (BHIP, GAMP, GMMP). High-performance liquid chromatography was used to quantify the relative metabolic rates of five major saponins under different culture conditions, and 16S rRNA sequencing was performed to analyze changes in microbial community structure. The results showed that the average relative metabolic rates of saponins R₁, Rb₁ and Rd in the BHIP group exceeded 73%; the GAMP group specifically and efficiently metabolized Rb₁, with a metabolic rate above 84%; while the GMMP group only metabolized R₁ and Re, with metabolic rates below 25%. Significant differences in microbial community structure were observed among the BHI, GAM, and GMM groups. Compared with their respective control groups, the α-diversity (Shannon index) of the microbiota in the BHIP and GAMP groups was significantly reduced (P < 0.01), and the community structure was significantly reshaped (P < 0.001), whereas no significant changes were observed in the GMMP group. In terms of microbial composition, the BHIP group showed significant enrichment of Lachnoclostridium, Parasutterella, and Bifidobacterium; the GAMP group was significantly enriched with Bifidobacterium and Escherichia-Shigella; and the GMMP group was mainly enriched with Roseburia and Anaerostipes. In conclusion, the interaction between PNS and the gut microbiota varies significantly under different culture conditions, and the environmental context should be fully considered when evaluating the probiotic effects of PNS.