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A New Rat Model of Pouchitis After Proctocolectomy and Ileal Pouch-Anal Anastomosis Using 2,4,6-Trinitrobenzene Sulfonic Acid

  • Original Article
  • Published:
Journal of Gastrointestinal Surgery

Abstract

Background

Pouchitis is a common complication after ileal pouch-anal anastomosis (IPAA) in patients with ulcerative colitis. However, an ideal model remains lacking. Therefore, we aimed to establish an appropriate model resembling human pouchitis.

Methods

Sprague-Dawley rats were randomly assigned to five groups: TNBS group, DSS group, NS group (following IPAA procedure, administrated with TNBS enema, DSS orally, normal saline enema, respectively), NI group (underwent IPAA), and sham group (underwent switch abdominal surgery). General status, weight change, hematochezia, and fecal scores were recorded. Fecal microbiota were counted under a microscope and analyzed by 16S rRNA gene high-throughput sequencing. Specimens of ileal pouch and small intestine (proximal, mid, distal) were collected to evaluate myeloperoxidase and occludin expression by immunohistochemistry and mRNA expression of pro-inflammatory markers by PCR.

Results

General status, hematochezia, fecal score, and increased mRNA expression of interleukin-6 and TNF-α in the TNBS group were similar to those in the DSS group, whereas the TNBS-induced model displayed a more stable weight change and more serious dysbacteriosis, not only was fecal bacterial diversity reduced, the dominant microbiota was altered. Histopathology scores of the distal small intestine in the TNBS group were lower compared with those in the DSS group (P < 0.05). A significant difference in myeloperoxidase and occludin expression in the small intestine was also detected between the TNBS and DSS groups.

Conclusions

Our model mimicked the characteristics of human pouchitis and avoided potential side effects in the small intestine, and thus could be employed for further research.

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Abbreviations

IPAA:

Ileal pouch-anal anastomosis

UC:

Ulcerative colitis

TNBS:

Trinitrobenzene sulfonic acid

DSS:

Dextran sulfate sodium

TNF-α:

Tumor necrosis factor-α

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Funding

This work was supported by the Li Jie-shou Gut Barrier Foundation (Grant No. LJS_201008).

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Author notes

  1. Kai-Yu Li and Xin Wang contributed equally to this paper and should be regarded as joint first authors

Authors and Affiliations

  1. Department of General Surgery, Tianjin Medical University General Hospital, Tianjin, 300052, China

    Kai-Yu Li, Xin Wang, Gang Liu, An-Qi He, Zi-Cheng Zheng, Xin-Yu Zhao & Tong Liu

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  1. Kai-Yu Li
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  3. Gang Liu
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Contributions

Kai-Yu Li and Xin Wang wrote the manuscript, processed the samples, analyzed the raw data, and approved the final version of the manuscript; Gang Liu conceived the study protocol, critically revised the manuscript, obtained funding, and approved its final version; An-Qi He, Zi-Cheng Zheng, Xin-Yu Zhao, and Tong Liu processed the samples, assessed and scored ileal-pouch and small intestinal histological damage, and approved the final version of the manuscript.

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Correspondence to Gang Liu.

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Li, KY., Wang, X., Liu, G. et al. A New Rat Model of Pouchitis After Proctocolectomy and Ileal Pouch-Anal Anastomosis Using 2,4,6-Trinitrobenzene Sulfonic Acid. J Gastrointest Surg 25, 1524–1533 (2021). https://doi.org/10.1007/s11605-020-04642-2

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