Regulating Effects of Dietary Fiber and Powder of Agaricus bisporus Based on in Vitro Fermentation on Human Gut Microbiota
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摘要: 目的：探究双胞菇膳食纤维以及双胞菇粉对肠道菌群的调节功能。方法：通过水提醇沉的方法提取双胞菇膳食纤维和双胞菇粉末进行肠道微生物体外发酵，测定代谢产物中pH、产气量、短链脂肪酸的含量，采用IlluminaPE250测序平台对粪便微生物V4区进行富集测序，探究双胞菇膳食纤维和双胞菇粉对于肠道菌群中短链脂肪酸产生菌相对丰度的影响。结果：以低聚果糖为阳性对照组，不加膳食纤维为空白对照组，体外发酵过程中，随着时间的增加，双胞菇膳食纤维组pH从6.93下降到4.48，双胞菇粉末pH从6.93下降至4.86；随着时间的增加，与空白组相比，双胞菇膳食纤维组和双胞菇粉末组的产气量分别增加了3.4和1.9 mL；在体外发酵24 h后发现，与阳性对照组相比，双胞菇膳食纤维组和双胞菇粉末组的肠道菌群丰富度和多样性更高；双胞菇膳食纤维组和双胞菇粉末组均能被肠道微生物利用产生短链脂肪酸，与空白组相比，双胞菇膳食纤维组更有利于被肠道微生物利用产生乙酸（34.3 mmol/L）和丙酸（7.6 mmol/L），促进有益菌属双歧杆菌属的生长，双胞菇粉末组更有利于产生乙酸（39.4 mmol/L），促进肠杆菌属的生长；与阳性对照组相比，双胞菇膳食纤维组和双胞菇粉末组的丙酸产量更高。结论：双胞菇膳食纤维和双胞菇粉末均能被肠道菌群有效利用，产生短链脂肪酸，提高有益菌的相对丰度，与空白组相比，添加双胞菇膳食纤维的实验组效果最佳，对人体肠道菌群存在有效的益生调节功能。Abstract: Objective: To explore the regulating function of the dietary fiber of Agaricus bisporus and the powder of Agaricus bisporus on the human gut microbiota. Methods: Dietary fiber of Agaricus bisporus were extracted by water extraction and alcohol precipitation for human gut microbial fermentation in vitro. The pH, gas production and short-chain fatty acid contents of metabolites were determined. The V4 region of fecal microorganisms was enriched and sequenced by IlluminaPE250 sequencing platform. Results: Taking fructo-oligosaccharide as the positive control group and without dietary fiber as the blank control group, during the in vitro fermentation process, with the increase of time, the pH value of the Agaricus bisporus dietary fiber group decreased from 6.93 to 4.48, and the pH value of the powder of Agaricus bisporus decreased from 6.93 to 4.86. With the increasing of time, compared with the blank group, the gas production of the Agaricus bisporus dietary fiber group and the powder of Agaricus bisporus group increased by 3.4 and 1.9 mL. After 24 hours of in vitro fermentation, it was found that compared with the positive control group, the richness and diversity of the human gut microbiota in the Agaricus bisporus dietary fiber group and the powder of Agaricus bisporus group were relatively higher. Compared with the blank group, dietary fiber of Agaricus bisporus group was more conducive to the production of acetic acid (34.3 mmol/L) and propionic acid (7.6 mmol/L) by human gut microbiota and promoted the growth of beneficial bacteria of Bifidobacterium. The powder of Agaricus bisporus group was more conducive to producing acetic acid (39.4 mmol/L) and promoting the growth of Enterobacter. Compared with the positive control group, the production of propionic acid in the Agaricus bisporus dietary fiber group and the powder of Agaricus bisporus group was relatively higher. Conclusion: Both the dietary fiber of the Agaricus bisporus and the powder of Agaricus bisporus could be effectively utilized by the human gut microbiota to produce short-chain fatty acids and increase the relative abundance of beneficial bacteria. Compared with blank group, dietary fiber of Agaricus bisporus added in experimental group had the best results, which had an effective prebiotic regulation function on human gut microbiota.
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图 2 发酵液样品在不同发酵时间的产气量
Figure 2. Gas production of fermentation samples at different fermentation times
图 3 发酵液样品在不同发酵时间的SCFAs值
Figure 3. SCFAS values of fermentation samples at different fermentation times
图 4 体外发酵样本Rank-Abundance曲线
Figure 4. Rank-Abundance curve of in vitro fermentation samples
图 5 体外发酵前后群落Alpha多样性分析
Figure 5. Alpha diversity of community before and after in vitro fermentation
图 6 体外发酵前后群落主坐标（PCoA）分析
Figure 6. PCoA analysis of community before and after in vitro fermentation
图 7 体外发酵24 h后肠道微生物群落组成变化
注：A.门水平；B.属水平。
Figure 7. Changes of intestinal microbial community composition after in vitro fermentation for 24 h
图 8 体外发酵24 h后Circos样本与物种关系图
Figure 8. Relationship between Circos samples and species after 24 h in vitro fermentation
表 1 短链脂肪酸各组分的线性回归方程
Table 1. Linear regression equation for components of short chain fatty acids
组分 线性回归方程 决定系数R2 乙酸 Y=18.602X−0.778 0.9999 丙酸 Y=31.342X−0.765 0.9998 丁酸 Y=41.565X−1.214 0.9999 
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