从多糖利用位点的差异表达研究两种健脾中药复方多糖影响脆弱拟杆菌体外生长机制

武妍凝; 吴莉; 伍荷洁; 舒青龙

doi:10.13422/j.cnki.syfjx.20230803

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从多糖利用位点的差异表达研究两种健脾中药复方多糖影响脆弱拟杆菌体外生长机制

经典名方 | 更新时间：2023-10-20

- 从多糖利用位点的差异表达研究两种健脾中药复方多糖影响脆弱拟杆菌体外生长机制
  增强出版
- Mechanism of Polysaccharides from Two Spleen-invigorating Chinese Medicine Prescriptions in Regulating Growth of Bacteroides fragilis in vitro： Based on Differential Expression of Polysaccharide Utilization Loci
- 中国实验方剂学杂志 2023年29卷第22期页码：21-28
- 作者机构：
  
  江西中医药大学中医学院，南昌 330004
- 作者简介：
  
  武妍凝，在读硕士，从事中医药微生态研究，E-mail：553154295@qq.com
  舒青龙，教授，从事中医药微生态研究，E-mail：shuqinglong@126.com
- 基金信息：
  
  国家自然科学基金项目(82160788);江西省卫生和计划生育委员会-科技计划项目(202211406);江西中医药大学创新创业训练计划项目(202210412172);江西中医药大学中西医结合一级学科（江西省双一流学科）项目(zxyylxk20220103)
- DOI：10.13422/j.cnki.syfjx.20230803
  中图分类号： R2-0;R22;R285.5;R289;R33
- 纸质出版日期：2023-11-20，
  
  网络出版日期：2023-03-24，
  
  收稿日期：2022-12-24，
- 稿件说明：
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武妍凝,吴莉,伍荷洁等.从多糖利用位点的差异表达研究两种健脾中药复方多糖影响脆弱拟杆菌体外生长机制[J].中国实验方剂学杂志,2023,29(22):21-28.

WU Yanning,WU Li,WU Hejie,et al.Mechanism of Polysaccharides from Two Spleen-invigorating Chinese Medicine Prescriptions in Regulating Growth of Bacteroides fragilisin vitro： Based on Differential Expression of Polysaccharide Utilization Loci[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(22):21-28.
武妍凝,吴莉,伍荷洁等.从多糖利用位点的差异表达研究两种健脾中药复方多糖影响脆弱拟杆菌体外生长机制[J].中国实验方剂学杂志,2023,29(22):21-28. DOI： 10.13422/j.cnki.syfjx.20230803.

WU Yanning,WU Li,WU Hejie,et al.Mechanism of Polysaccharides from Two Spleen-invigorating Chinese Medicine Prescriptions in Regulating Growth of Bacteroides fragilisin vitro： Based on Differential Expression of Polysaccharide Utilization Loci[J].Chinese Journal of Experimental Traditional Medical Formulae,2023,29(22):21-28. DOI： 10.13422/j.cnki.syfjx.20230803.

摘要

目的

探讨两种中药复方多糖作为碳源对脆弱拟杆菌生长的影响及其基于转录组学从多糖利用位点差异表达角度的机制分析。

方法

使用不同的碳源培养基（分别以浓度为20%的理中汤多糖和参苓白术散多糖、葡萄糖、脑心浸液（BHI）为唯一碳源）体外厌氧培养脆弱拟杆菌（

Bacteroides fragile

ATCC25285），通过连续取样法和分光光度法测定不同碳源对脆弱拟杆菌ATCC25285的生长影响。再对不同碳源培养基的脆弱拟杆菌ATCC25285进行转录组测序与分析，研究不同碳源影响细菌生长的机制。

结果

不同碳源体外培养结果显示，理中汤多糖组、参苓白术散多糖组、BHI组和葡萄糖组分别在26、32、26、38 h细菌生长达到细菌浓度最大值，均可以得到良好生长。转录组测序结果显示，基因本体论（GO）富集表明理中汤多糖组与参苓白术散多糖组集中在二羧酸转运和二羧酸跨膜转运活性等功能条目，多糖利用位点（PULs）差异分析表明参苓白术散多糖组与BHI组比较，两者共同高表达了PUL（4、27）及糖苷水解酶13（GH13）和糖基转移酶5（GT5），参苓白术散组单独高表达了PUL9，BHI组单独高表达PUL（17、19、20）及GH3、GH144；参苓白术散多糖组与葡萄糖组相比，PUL27及GT5在两组中均高表达，参苓白术散多糖组中单独高表达PUL（4、9）及GH13，葡萄糖组中单独高表达PUL（2、17、19）及GH2；理中汤多糖组与BHI组比较，两者共同显著表达PUL（4、17、19、20、27）及GH3、GH144，理中汤多糖组中单独高表达PUL（2、8、23、27）及GH2、GH57，BHI组中单独升高表达的为GH13；理中汤组与葡萄糖组比较，两者共同显著表达了PUL（4、27）及GH2，理中汤多糖组中单独高表达PUL（4、8、20、23）及GH3、GH144，葡萄糖组单独高表达PUL30。

结论

不同碳源对脆弱拟杆菌ATCC25285的体外实验与转录组结果相互印证，PULs和GH的表达可能根据不同碳源的营养情况对脆弱拟杆菌ATCC25285适应性生长提供益处或代价，这或许是中药复方多糖调节脆弱拟杆菌ATCC25285的生长机制之一，可为深入研究脆弱拟杆菌代谢途径与中药复方多糖之间的联系提供参考。

Abstract

Objective

To investigate the effects of using the polysaccharides from two Chinese medicine compound prescriptions as the carbon source on the growth of

Bacteroides fragilis

and to decipher the mechanism from the perspective of differential expression of polysaccharide utilization loci （PULs） based on transcriptomics.

Method

The media with different carbon sources ［20% polysaccharides of Lizhongtang， polysaccharides of Shenling Baizhusan， glucose， and brain heart infusion （BHI） Broth］ were used for the anaerobic culture of

B. fragile

ATCC25285. The effects of different carbon sources on the growth of

B. fragilis

ATCC25285 were determined by continuous sampling and spectrophotometry. Then， transcriptome sequencing was performed for the cultures obtained with different carbon sources to study the mechanism of different carbon sources in regulating bacterial growth.

Result

The concentration of bacteria with the polysaccharide of Lizhongtang， polysaccharide of Shenling Baizhusan， BHI Broth， and glucose as the carbon sources peaked at 26， 32， 26， 38 h， respectively， and the bacteria in all the four groups achieved robust growth. Gene ontology （GO） enrichment indicated that the differentially expressed genes in the Lizhongtang polysaccharide group and Shenling Baizhusan polysaccharide group were concentrated in the transport and transmembrane transport of dicarboxylic acid. The Shenling Baizhusan polysaccharide and BHI Broth groups showed high expression of PUL 4 and 27， glycoside hydrolase 13 （GH13）， and glycosyl transferases 5 （GT5）. PUL9 was highly expressed in Shenling Baizhusan polysaccharide group， and PUL 17， 19， and 20， GH3， and GH144 in the BHI Broth group. PUL27 and GT5 were highly expressed in Shenling Baizhusan polysaccharide and glucose groups. PUL 4 and 9 and GH13 were only highly expressed in Shenling Baizhusan polysaccharide group， and PUL 2， 17， and 19 and GH2 in the glucose group. Both Lizhongtang polysaccharide group and BHI group highly expressed PUL 4， 17， 19， 20， and 27， GH3， and GH144. PUL 2， 8， 23， and 27， GH2， and GH57 were highly expressed in Lizhongtang polysaccharide group， while GH13 showed high expression in the BHI group. Both the glucose and Lizhongtang polysacharride groups showed high expression of PUL 4 and 27 and GH2. PUL 4， 8， 20， and 23， GH3， and GH144 were highly expressed in Lizhongtang polysaccharide group， while PUL30 was highly expressed in the glucose group.

Conclusion

The

in vitro

experiments and transcriptome sequencing results confirmed that the expression of PULs and GH may provide benefits or costs to the adaptive growth of

Bacteroides fragilis

ATCC25285 cultured with different carbon sources， which may be one of the mechanisms by which polysaccharides from Chinese medicine compound prescriptions regulate the growth of

B. fragilis

ATCC25285. The findings can provide a reference for further research on the relationship between

B. fragilis

metabolic pathway and polysaccharides of Chinese medicine compound prescriptions.

关键词

理中汤参苓白术散碳源脆弱拟杆菌转录组多糖利用位点

Keywords

LizhongtangShenling Baizhusancarbon sourceBacteroides fragilis ATCC25285transcriptomepolysaccharide utilization loci

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