实时荧光定量PCR联合高通量测序分析杠柳毒苷与三七总皂苷配伍对大鼠肠道菌群的影响

目的 采用实时荧光定量PCR （qRT-PCR）法和Illumina高通量测序技术研究杠柳毒苷和不同比例三七总皂苷配伍对大鼠肠道菌群的影响。方法 将15只SD大鼠随机分为杠柳毒苷单独给药组（A组）、杠柳毒苷和低剂量三七总皂苷配伍组（B组）、杠柳毒苷和高剂量三七总皂苷配伍组（C组），每组5只。按杠柳毒苷10 mg/kg、三七总皂苷0.74和2.22 g/kg （分别相当于香加皮与三七的配伍比例为1：1和1：3）的剂量ig给药，连续给药7 d。给药结束后收集大鼠粪便，进行qRT-PCR反应和高通量测序。结果 qRT-PCR结果表明，与杠柳毒苷单独给药组比较，杠柳毒苷和三七总皂苷配伍后，总菌、拟杆菌的相对含量显著升高（P<0.05、0.01），而乳酸杆菌的相对含量呈现降低的趋势；Illumina高通量测序结果显示，在微生物群落结构上，杠柳毒苷配伍三七总皂苷后，拟杆菌的相对丰度明显升高而乳酸杆菌的相对丰度呈现降低的趋势；在肠道菌群多样性上，3组样品的多样性指数间无显著性差异。结论 杠柳毒苷和不同比例的三七总皂苷配伍后，可对大鼠肠道中的总菌、乳酸杆菌及拟杆菌产生一定影响，但对肠道菌群多样性的影响不明显。

Compatibility of periplocin and Panax notoginseng saponins on intestinal flora in SD rats by Real-time fluorescence quantitative PCR and High-throughput sequencing

Objective To study the influence of combination of Panax notoginseng saponins (PNS) and periplocin on intestinal flora in rats by using Illumina high-throughput sequencing and Real-time fluorescence quantitative PCR (qRT-PCR).Methods Fifteen SD rats were randomly divided into three groups:the periplocin single administration group (group A),compatibility group of periplocin and low dose PNS (group B),compatibility group of periplocin and high dose PNS (group C).Periplocin 10 mg/kg,PNS 0.74 and 2.22 g/kg (corresponding to the compatibility ratio of Periplocae Cortex and Panax notoginseng 1:1 and 1:3,respectively) were administered by ig for seven days.After seven days of administration,the fecal samples were collected and analyzed by qRT-PCR and Illumina high-throughput sequencing.Results qThe qRT-PCR results showed that the bacterium number of total gut bacteria and Bacteroidetes were significantly increased (P< 0.05 and 0.01),while the relative content of Lactobacillus was decreased in the combination group,compared with Periplocin group.Illumina high-throughput sequencing results showed that,in terms of microbial community structure,the relative abundance of Bacteroides was increased significantly and that of Lactobacillus was decreased after the combination of periplocin and PNS,and there was no significant difference in the diversity index of intestinal flora among three groups.Conclusion The compatibility of periplocin and PNS in different ratios has some effect on gut flora of rats to some extent,but has little effect on diversity of intestinal flora.