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四君子汤均一多糖对体外人肠道微生物及短链脂肪酸的影响。

Effects of a homogeneous polysaccharide from Sijunzi decoction on human intestinal microbes and short chain fatty acids in vitro.

机构信息

School of Pharmacy, Shanghai Jiao Tong University, No. 800 Dongchuan Road, Minhang District, Shanghai 200240, China.

School of Pharmacy, Shanghai Jiao Tong University, No. 800 Dongchuan Road, Minhang District, Shanghai 200240, China.

出版信息

J Ethnopharmacol. 2018 Oct 5;224:465-473. doi: 10.1016/j.jep.2018.06.006. Epub 2018 Jun 15.

Abstract

ETHNOPHARMACOLOGICAL RELEVANCE

Sijunzi decoction (SJZD) is a classic recipe in traditional Chinese medicine (TCM) to strengthen the spleen and replenish Qi. It is well known for treating disorders of gastrointestinal function manifested in poor appetite, reduced food intake and loose stools. Polysaccharide is the most abundant constituent and the major effective component in SJZD.

AIM OF THE STUDY

The present study aimed to understand the immunomodulatory mechanism of S-3-1, a homogeneous polysaccharide purified from SJZD with immune-enhancement activity, by investigating its effects on human intestinal microbes and short chain fatty acids.

MATERIALS AND METHODS

S-3-1 was incubated with simulated gastric juice, intestinal juice, and human fecal microflora independently and sequentially. The concentrations of total polysaccharide and reducing sugar were measured to identify the stability of independently and sequentially incubated S-3-1 in three in vitro fermentation models. Gas chromatograph (GC) analysis was used to measure the short chain fatty acid (SCFA) contents in human fecal samples. The human gut microbiota composition was measured by 16S rRNA gene Illumina MiSeq sequencing (V3-V4 region).

RESULTS

S-3-1 was degraded in three in vitro fermentation models separately and sequentially. Both S-3-1 and incubated S-3-1 could regulate the abundances of Lactobacillus, Pediococcus, Streptococcus, Bacteroides, Enterococcus, Clostridium and Dorea in human intestinal microflora samples. Specifically, S-3-1 could only regulate the abundances of Paraprevotella and Oscillospira, while the influenced flora changed to Butyricimonas, Coprococcus, Dialister, Sutterella, Ruminococcus and Parabacteroides after sequential incubation of S-3-1. In contrast to S-3-1 showing no influence on the content of SCFA, incubated S-3-1 showed increased contents of acetic acid and total acid that were associated with its effects on the abundances of Enterococcus, Sutterella, Butyricimonas and Streptococcus.

CONCLUSION

S-3-1 plays an immunomodulatory role by regulating the abundances of 9 intestinal bacteria genera. Incubated S-3-1 can regulate more bacteria genera, a total of 13 kinds, and can adjust the SCFA content to affect immunomodulation. Incubation with gastric and intestinal juices enhanced S-3-1's capability of modulating the intestinal flora composition and decreased the bacteria's need for a carbon source. This study could provide new insights for studies on the pharmacological mechanisms of polysaccharides in vitro.

摘要

民族药理学相关性

四君子汤(SJZD)是一种经典的中药配方,用于增强脾脏和补充气。它以治疗胃肠道功能障碍为特征,表现为食欲不振、食量减少和腹泻。多糖是 SJZD 中最丰富的成分和主要有效成分。

研究目的

本研究旨在通过研究其对人类肠道微生物和短链脂肪酸的影响,了解 S-3-1(一种具有免疫增强活性的 SJZD 均一多糖)的免疫调节机制。

材料和方法

S-3-1 分别与模拟胃液、肠液和人粪便微生物群依次孵育。测量总多糖和还原糖的浓度,以鉴定三种体外发酵模型中独立和连续孵育的 S-3-1 的稳定性。气相色谱(GC)分析用于测量人粪便样本中的短链脂肪酸(SCFA)含量。通过 16S rRNA 基因 Illumina MiSeq 测序(V3-V4 区)测量人类肠道微生物群落组成。

结果

S-3-1 在三种体外发酵模型中分别和连续孵育时均被降解。S-3-1 和孵育的 S-3-1 均可调节人类肠道微生物群样本中乳杆菌、肠球菌、链球菌、拟杆菌、粪肠球菌、梭菌和 Dorea 的丰度。具体而言,S-3-1 只能调节 Paraprevotella 和 Oscillospira 的丰度,而连续孵育 S-3-1 后,受影响的菌群变为 Butyricimonas、Coprococcus、Dialister、Sutterella、Ruminococcus 和 Parabacteroides。与 S-3-1 对 SCFA 含量没有影响相反,孵育的 S-3-1 显示出乙酸和总酸含量增加,这与其对肠球菌、Sutterella、Butyricimonas 和链球菌丰度的影响有关。

结论

S-3-1 通过调节 9 种肠道细菌属的丰度发挥免疫调节作用。孵育的 S-3-1 可以调节更多的细菌属,共 13 种,并且可以调节 SCFA 含量以影响免疫调节。与胃液和肠液孵育增强了 S-3-1 调节肠道菌群组成的能力,并降低了细菌对碳源的需求。本研究可为多糖的体外药理学机制研究提供新的见解。

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