Study on the Effect of Intestinal Flora on Intestinal Motility in Rats with Slow Transit Constipation of <italic style="font-style: italic">Qi </italic>Stagnation Pattern Based on Pseudo-Sterile Experiment and Fecal Microbiota Transplantation

LIU Qihong; KE Xiao; LUO Yunfeng; HU Lunan; REN Yan; FANG Wenyi; ZHAO Peilin; YAN Jinxian

doi:10.13288/j.11-2166/r.2024.09.012

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Study on the Effect of Intestinal Flora on Intestinal Motility in Rats with Slow Transit Constipation of Qi Stagnation Pattern Based on Pseudo-Sterile Experiment and Fecal Microbiota Transplantation

更新时间：2024-04-25

- Study on the Effect of Intestinal Flora on Intestinal Motility in Rats with Slow Transit Constipation of Qi Stagnation Pattern Based on Pseudo-Sterile Experiment and Fecal Microbiota Transplantation
- Journal of Traditional Chinese Medicine Vol. 65, Issue 9, Pages: 943-948(2024)
- 作者机构：
  
  1.福建中医药大学附属第二人民医院，福建省福州市五四路282号，350003
  2.福建省医疗机构中药制剂重点实验室
- 作者简介：
- 基金信息：
- DOI：10.13288/j.11-2166/r.2024.09.012
  CLC：
- Received：17 November 2023，
  
  Revised：2024-02-09，
  
  Published：02 May 2024
- 稿件说明：
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刘启鸿,柯晓,骆云丰等.基于伪无菌实验与肠菌移植技术研究气滞证慢传输型便秘模型大鼠肠道菌群对肠道动力的影响[J].中医杂志,2024,65(09):943-948.

LIU Qihong,KE Xiao,LUO Yunfeng,et al.Study on the Effect of Intestinal Flora on Intestinal Motility in Rats with Slow Transit Constipation of Qi Stagnation Pattern Based on Pseudo-Sterile Experiment and Fecal Microbiota Transplantation[J].Journal of Traditional Chinese Medicine,2024,65(09):943-948.
刘启鸿,柯晓,骆云丰等.基于伪无菌实验与肠菌移植技术研究气滞证慢传输型便秘模型大鼠肠道菌群对肠道动力的影响[J].中医杂志,2024,65(09):943-948. DOI： 10.13288/j.11-2166/r.2024.09.012.

LIU Qihong,KE Xiao,LUO Yunfeng,et al.Study on the Effect of Intestinal Flora on Intestinal Motility in Rats with Slow Transit Constipation of Qi Stagnation Pattern Based on Pseudo-Sterile Experiment and Fecal Microbiota Transplantation[J].Journal of Traditional Chinese Medicine,2024,65(09):943-948. DOI： 10.13288/j.11-2166/r.2024.09.012.

摘要

目的

运用伪无菌实验与肠菌移植技术，探讨气滞证慢传输型便秘（STC）大鼠的肠道菌群对肠道动力的影响以及二者的关系。

方法

24只Wistar大鼠随机分为正常组6只、气滞证STC组6只、伪无菌组12只。气滞证STC组采用洛哌丁胺混悬液3 mg/kg灌胃联合夹尾刺激建立气滞证STC大鼠模型，造模成功后分别采集气滞证STC组和正常组大鼠的新鲜粪便制备100 mg/ml粪菌混悬液。伪无菌组采用抗生素鸡尾酒法（给予含杆菌肽、硫酸链霉素、硫酸新霉素各20 mg/ml的混合抗生素混悬液灌胃）建立伪无菌大鼠模型，造模成功后将大鼠随机分为正常粪菌液组、气滞证STC粪菌液组各6只，再分别给予制备的相应大鼠粪菌混悬液10 ml/kg灌胃；气滞证STC组大鼠给予等体积无菌水灌胃，均每日1次、连续7天。比较气滞证STC组、正常粪菌液组、气滞证STC粪菌液组小肠推进率；ELISA法检测血清5-羟色胺（5-HT）水平；免疫组化法检测结肠组织5-羟色胺3受体（5-HT3R）、5-羟色胺4受体（5-HT4R）阳性表达水平；Western blot法检测结肠组织色氨酸羟化酶1（TPH1）、色氨酸羟化酶2（TPH2）、5-羟色胺转运体（SERT）、单胺氧化酶A（MAO-A）蛋白表达水平。

结果

与正常粪菌液组比较，气滞证STC组、气滞证STC粪菌液组大鼠小肠推进率、血清5-HT水平，结肠组织5-HT3R、5-HT4R阳性表达以及TPH1、TPH2、SERT、MAO-A蛋白表达均降低（

＜0.05）；气滞证STC组和气滞证STC粪菌液组各项指标比较差异均无统计学意义（

＞0.05）。

结论

气滞证STC大鼠的肠道菌群紊乱可导致肠道传输功能减慢，其机制可能是通过影响5-HT的合成、转运、代谢等途径而导致肠道动力障碍。

Abstract

Objective

To clarify the relationship between intestinal flora and intestinal motility in rats with slow transit constipation （STC） and

stagnation syndrome by conducting a pseudo-sterile experiment and fecal microbiota transplantation （FMT） technology.

Methods

Twenty-four Wistar rats were randomly divided into normal group （n=6）， STC with

stagnation pattern group （n=6） and pseudo-sterile group （n=12）. In the STC group with

stagnation pattern， 3 mg/kg of loperamide suspension by intragastric administration combined with tail clamping stimulation were performed to establish the rat model of STC with

stagnation pattern. After successful modeling， fresh feces from the rats in the STC with

stagnation pattern group and the normal group were collected to prepare 100 mg/ml of fecal bacterial suspension. In the pseudo-sterile group， the antibiotic cocktail method was used （a mixed antibiotic suspension containing bacitracin， streptomycin sulfate， and neomycin sulfate at 20 mg/ml each was administered intragastrically） to establish pseudo-sterile rats model. After successful modeling， the rats were randomly divided into normal fecal bacterial liquid group and STC with

stagnation pattern fecal bacterial liquid group， with six rats in each group， and then were given 10 ml/kg of the prepared corresponding rat fecal bacterial suspension by gavage. Rats in STC with

stagnation pattern group were given an equal volume of sterile water by gavage. All groups were administered once a day for 7 consecutive days. The small intestinal propulsion rate of the STC with

stagnation pattern group， the normal fecal bacterial liquid group， and STC with

stagnation pattern fecal bacterial liquid group were compared. ELISA method was used to detect serum 5-hydroxytryptamine （5-HT） levels. Immunohistochemistry was used to detect the positive expression levels of 5-hydroxytryptamine 3 receptor （5-HT3R） and 5-hydroxytryptamine 4 receptor （5-HT4R） in colon tissue. Western blot method was used to detect the protein expression levels of tryptophan hydroxylase 1 （TPH1）， tryptophan hydroxylase 2 （TPH2）， serotonin transporter （SERT）， and monoamine oxidase A （MAO-A） in colon tissue.

Results

Compared to those in the normal fecal bacterial liquid group， the small intestinal propulsion rate， serum 5-HT level， positive expression of 5-HT3R and 5-HT4R in colon tissue， and protein expression of TPH1， TPH2， SERT and MAO-A significantly decreased in the STC with

stagnation pattern group and STC with

stagnation pattern fecal bacterial liquid group （

＜0.05）. There was no statistically significant difference in the indicators between the STC with

stagnation pattern group and STC with

stagnation pattern fecal bacterial liquid group （

＞0.05）.

Conclusion

The intestinal flora in STC rats with

stagnation pattern can lead to a slowdown in intestinal transmission function， whose mechanism may be related to intestinal motility disorders affected by the synthesis， transport， metabolism and other pathways of 5-HT.

关键词

Keywords

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Study on the Effect of Intestinal Flora on Intestinal Motility in Rats with Slow Transit Constipation of Qi Stagnation Pattern Based on Pseudo-Sterile Experiment and Fecal Microbiota Transplantation

DOI：10.13288/j.11-2166/r.2024.09.012

摘要

Abstract

关键词

Keywords

references