Effects of <italic style="font-style: italic">Chaihu Shugan Powder</italic> （柴胡疏肝散） on Duodenal Mucosal Barrier and Intestinal Flora in Functional Dyspepsia Model Rats

YANG Keming; JIA Yuebo; YELIYA Yeertai; LIU Xuejiao; WANG Xiangxiang; JIANG Kailin; LING Jianghong

doi:10.13288/j.11-2166/r.2023.23.012

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Effects of Chaihu Shugan Powder （柴胡疏肝散） on Duodenal Mucosal Barrier and Intestinal Flora in Functional Dyspepsia Model Rats

更新时间：2023-11-24

- Effects of Chaihu Shugan Powder （柴胡疏肝散） on Duodenal Mucosal Barrier and Intestinal Flora in Functional Dyspepsia Model Rats
- Journal of Traditional Chinese Medicine Vol. 64, Issue 23, Pages: 2443-2453(2023)
- 作者机构：
  
  上海中医药大学附属曙光医院，上海市普安路185号，201203
- 作者简介：
- 基金信息：
- DOI：10.13288/j.11-2166/r.2023.23.012
  CLC：
- Received：25 July 2023，
  
  Revised：2023-09-14，
  
  Published：02 December 2023
- 稿件说明：
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杨珂鸣,贾越博,叶丽亚·叶尔太等.柴胡疏肝散对功能性消化不良模型大鼠十二指肠黏膜屏障和肠道菌群的影响[J].中医杂志,2023,64(23):2443-2453.

YANG Keming,JIA Yuebo,YELIYA Yeertai,et al.Effects of Chaihu Shugan Powder （柴胡疏肝散） on Duodenal Mucosal Barrier and Intestinal Flora in Functional Dyspepsia Model Rats[J].Journal of Traditional Chinese Medicine,2023,64(23):2443-2453.
杨珂鸣,贾越博,叶丽亚·叶尔太等.柴胡疏肝散对功能性消化不良模型大鼠十二指肠黏膜屏障和肠道菌群的影响[J].中医杂志,2023,64(23):2443-2453. DOI： 10.13288/j.11-2166/r.2023.23.012.

YANG Keming,JIA Yuebo,YELIYA Yeertai,et al.Effects of Chaihu Shugan Powder （柴胡疏肝散） on Duodenal Mucosal Barrier and Intestinal Flora in Functional Dyspepsia Model Rats[J].Journal of Traditional Chinese Medicine,2023,64(23):2443-2453. DOI： 10.13288/j.11-2166/r.2023.23.012.

摘要

目的

探讨柴胡疏肝散治疗功能性消化不良的可能作用机制。

方法

将24只SD大鼠随机分为正常组、模型组、柴胡疏肝散组和益生菌组，每组6只。除正常组外，其余各组大鼠均予改良夹尾激惹法复制功能性消化不良大鼠模型。造模同时，正常组及模型组予生理盐水10 ml/（kg·d）灌胃，柴胡疏肝散组予柴胡疏肝散水煎液9.6 g/（kg·d）灌胃，益生菌组予益生菌水溶液0.945 g/（kg·d）灌胃，每天2次，连续4周。4周后检测各组大鼠胃排空率与小肠推进率，HE染色观察大鼠胃窦及十二指肠组织病理学变化，16s rDNA高通量基因测序法分析大鼠的肠道菌群变化，并且采用免疫组织化学法及蛋白免疫印迹法检测十二指肠带状闭合蛋白1（ZO-1）和咬合蛋白（Occludin）的表达，并对肠道菌群与ZO-1、Occludin蛋白表达进行Pearson相关性分析。

结果

各组大鼠胃窦组织结构层次清晰，腺体结构规整，胃组织黏膜光滑，未见糜烂、溃疡等病理改变。与正常组比较，模型组十二指肠组织肠绒毛明显减少或萎缩，杯状细胞排列紊乱，可见嗜酸性粒细胞浸润，大鼠胃排空率及小肠推进率、十二指肠组织中ZO-1、Occludin蛋白表达均降低（

＜0.01）。与模型组比较，柴胡疏肝散组及益生菌组十二指肠组织结构层次清晰，肠绒毛长度较长，杯状细胞排列较为整齐，柴胡疏肝散组未见明显嗜酸性粒细胞浸润，大鼠胃排空率、小肠推进率及ZO-1、Occludin蛋白表达均升高，益生菌组可见少量嗜酸性粒细胞浸润，大鼠胃排空率、Occludin蛋白表达升高（

＜0.05或

＜0.01）。肠道菌群Beta多样性分析显示，与正常组比较，模型组的肠道菌群整体结构出现了显著变化（

＜0.01）。与模型组比较，柴胡疏肝散组和益生菌组的肠道菌群整体结构更接近正常组。物种组成分析，与正常组比较，模型组厚壁菌门相对丰度降低，拟杆菌门、未分类的

Muribaculaceae

菌属相对丰度增加，拟杆菌门/厚壁菌门值升高（

＜0.05或

＜0.01）。与模型组比较，柴胡疏肝散组和益生菌组厚壁菌门相对丰度增加，拟杆菌门、未分类的

Muribaculaceae

菌属相对丰度降低，拟杆菌门/厚壁菌门值降低（

＜0.05或

＜0.01）。益生菌组与柴胡疏肝散组各指标组间比较差异均无统计学意义（

＞0.05）。Pearson相关性分析显示，在门水平上，厚壁菌门与ZO-1（

=0.610，

=0.016）、Occludin（

=0.694，

=0.004）蛋白表达量呈正相关。拟杆菌门与ZO-1（

0.557，

=0.031）、Occludin（

0.662，

=0.007）蛋白表达量呈负相关。在属水平上，未分类的

Muribaculaceae

菌属与ZO-1（

0.727，

=0.002）、Occludin（

0.760，

=0.001）蛋白表达量呈负相关。

结论

柴胡疏肝散可能通过恢复肠道菌群结构，调节厚壁菌门、拟杆菌门和未分类的

Muribaculaceae

菌属的丰度水平，同时上调ZO-1、Occludin蛋白，进而修复十二指肠黏膜屏障，对功能性消化不良起到治疗作用。

Abstract

Objective

To study the possible mechanism of

Chaihu Shugan Powder

（柴胡疏肝散， CSP） in the treatment of functional dyspepsia （FD）.

Methods

Twenty-four SD rats were randomly divided into a normal group， a model group， a CSP group and a probiotic group， with six rats in each group.The tail-clamping provocation method was used in all groups except for the normal group to replicate the FD rat model. Simultaneously， the normal group and the model group were given 10 ml/（kg·d） of saline by gavage， while the CSP group and the probiotic group were given 9.6 g/（kg·d） of CSP aqueous decoction and 0.945 g/（kg·d） of probiotic aqueous solution by gavage， respectively， twice daily for four weeks. After four weeks， the gastric emptying and small intestinal propulsion rates were detected in each group of rats. Hematoxylin-eosin （HE） staining was used to observe the histopathological changes in the gastric sinusoids and duodenum of the rats. The changes in the intestinal flora were analyzed by 16s rDNA high-throughput gene sequencing， and the expressions of the duodenal zona occludin 1 （ZO-1） and Occludin were detected by immunohistochemistry and western blotting. Pearson correlation analysis was performed on intestinal flora and ZO-1 and Occludin protein expression.

Results

The gastric antrum tissue structure was clear in all groups， and the gland structure was regular， with smooth gastric tissue mucosa and no pathological changes such as erosion and ulcer. Compared to those in the normal group， the intestinal villi in the duodenal tissue in the model group were significantly reduced or atrophied， and the goblet cells were arranged in disorder， with eosinophilic infiltration； the gastric emptying rate and small intestinal propulsion rate， as well as ZO-1 and Occludin protein expression in duodenal tissue significantly decreased （

＜0.01）. Compared to those in the model group， the duodenal tissue structure was clear， and the length intestinal villi was longer， with goblet cells neatly arranged in the CSP group and the probiotic group； no obvious eosinophil infiltration was found， and the gastric emptying rate and small intestinal propulsion rate as well as ZO-1 and Occludin protein expression significantly increased in the CSP group； a small amount of eosinophil infiltration was found， and the gastric emptying rate and Occludin protein expression significantly increased in the probiotic group （

＜0.05 or

＜0.01）. Beta diversity analysis of intestinal flora showed that the overall structure of intestinal flora in the model group changed significantly compared to that in the normal group （

＜0.01）. The overall structure of the intestinal flora in the CSP group and the probiotic group was closer to the normal group than the model group. Species composition analysis showed that the relative abundance of the

Firmicutes

decreased， while the relative abundance of the

Bacteroidetes

and

norank_f_Muribaculaceae

increased， and the

Bacteroidetes/Firmicutes

value increased in the model group than those in the normal group （

＜0.05 or

＜0.01）. Compared to those in the model group， the relative abundance of the

Firmicutes

increased， while the relative abundance of the

Bacteroidetes

and

norank_f_Muribaculaceae，

as well as

the

Bacteroidetes/Firmicutes

value decreased in the CSP group and the probiotic group （

＜0.05 or

＜0.01）. There was no statistically significant difference in each indicator between the probiotic group and the CSP group （

＞0.05）. Pearson correlation analysis showed that at the phylum level，

Firmicutes

was positively correlated with ZO-1 （

=0.610，

=0.016） and Occludin （

=0.694，

=0.004） protein expression.

Bacteroidetes

was negatively correlated with ZO-1 （

0.557，

=0.031） and Occludin （

0.662，

=0.007） protein expression. At the genus level，

norank_f_Muribaculaceae

was negatively correlated with ZO-1 （

0.727，

=0.002） and Occludin （

0.760，

=0.001） protein expression.

Conclusion

CSP can restore the structure of intestinal flora， regulate the abundance levels of

Firmicutes

，

Bacteroidetes

and

norank_f_Muribaculaceae

， up-regulate ZO-1 and Occludin proteins， and thus repairing the duodenal mucosal barrier， and playing a therapeutic role in FD rats.

关键词

Keywords

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⁰

Effects of Chaihu Shugan Powder （柴胡疏肝散） on Duodenal Mucosal Barrier and Intestinal Flora in Functional Dyspepsia Model Rats

DOI：10.13288/j.11-2166/r.2023.23.012

摘要

Abstract

关键词

Keywords

references