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Induction of Murine TNBS Colitis Is Strictly Controlled by a Modified Method Using Continuous Inhalation Anesthesia with Sevoflurane

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Abstract

Background

Trinitrobenzenesulfonic acid (TNBS)-induced colitis is one of the most widely used experimental colitis models. However, there is no standard procedure for inducing colitis by TNBS because it is difficult to achieve a uniform distribution of colitis. We have developed a modified method of murine TNBS-induced colitis that involves inhalation anesthesia with sevoflurane combined with both single and repeated TNBS administrations.

Aims

To compare the usefulness of our newly developed method for inducing murine TNBS-induced colitis with that of conventional intraperitoneal anesthesia.

Methods

TNBS in ethanol was administered to C57BL/6J mice held in an inverted vertical position either under continuous inhalation anesthesia with sevoflurane, in accordance with our newly developed method, or by intraperitoneal injection with 2.5 % avertin, in accordance with the conventional procedure. Body weight change, cytokine profile, and histological findings were examined during the course of colitis.

Results

The dispersion of anesthesia time, TNBS retention time, and nadir weight during the course of colitis was decreased using the newly developed method compared with the conventional procedure. Optimization of the modified TNBS-induced colitis, as evidenced by the predominant expression of Th1 and Th17 cytokines on day 7, was attained by the injection of 2.25 mg TNBS in 55 % ethanol. Regulation of the TNBS retention time using inhalation anesthesia with sevoflurane allowed strict control of the disease severity of TNBS-induced colitis. Using the modified method we were also able to develop a chronic TNBS-induced colitis model by repeated TNBS administration without excessive mortality of the mice.

Conclusions

Our modified method for murine TNBS-induced colitis using continuous inhalation anesthesia with sevoflurane provides a better experimental colitis model following both single and repeated TNBS administrations.

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Abbreviations

CIA:

Continuous inhalation anesthesia

IBD:

Inflammatory bowel disease

IFN-γ:

Interferon gamma

IL:

Interleukin

IPA:

Intraperitoneal anesthesia

Th:

T helper

TNBS:

Trinitrobenzenesulfonic acid

TNF-α:

Tumor necrosis factor alpha

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Acknowledgments

This research was supported by Grant-in-Aid for Scientific Research (C) 21590808 and 25460948 from the Ministry of Education, Culture, Sports, Science and Technology, Japan. We are grateful to Soichiro Mimuro for technical assistance regarding anesthesia in mice.

Conflict of interest

None.

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Authors and Affiliations

  1. First Department of Medicine, Hamamatsu University School of Medicine, Hamamatsu, 431-3192, Japan

    Tomohiro Terai, Shinya Tani, Shinji Oishi, Takanori Yamada, Mitsushige Sugimoto, Shigeru Kanaoka, Hiroaki Miyajima & Ken Sugimoto

  2. Department of Endoscopic and Photodynamic Medicine, Hamamatsu University School of Medicine, 1-20-1 Handayama, Higashi-ku, Hamamatsu, 431-3192, Japan

    Satoshi Osawa

  3. Department of Pathology, Seirei Hamamatsu General Hospital, Hamamatsu, 430-8558, Japan

    Yoshifumi Arai

  4. Center for Clinical Research, Hamamatsu University School of Medicine, Hamamatsu, 431-3192, Japan

    Takahisa Furuta

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  1. Tomohiro Terai
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  2. Satoshi Osawa
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Correspondence to Satoshi Osawa.

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10620_2013_3023_MOESM2_ESM.tif

Supplemental Fig. 1 Evaluation of single TNBS administration in BALB/c mice using continuous inhalation anesthesia. Female BALB/c mice were anesthetized by continuous ventilation with sevoflurane, and TNBS colitis was induced according to the protocol. On day 0, 1.25 mg of TNBS in 50 % ethanol was administered (n = 12). (a) Time course of body weight change during colitis. (b) Time course of TNBS retention volume. ©) Hematoxylin and eosin staining and Masson trichrome staining of representative colon sections on day 0 (control), 3, and 7 after TNBS administration. 10× magnification in upper section; 40× magnification in lower section. Histological findings on day 3 showed distortion of crypts, loss of goblet cells, and focal ulcers accompanied by edema in the mucosal or submucosal regions. By day 7, disrupted crypts and goblet cells regenerated in most parts of the mucosa, but infiltration of mononuclear cells was still prominent in the stromal tissue of basal mucosal areas. Trichrome-stained collagen deposits were observed in the stromal tissue in the mucosa and submucosa on day 3 and 7, respectively. (TIFF 2,885 kb)

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Terai, T., Osawa, S., Tani, S. et al. Induction of Murine TNBS Colitis Is Strictly Controlled by a Modified Method Using Continuous Inhalation Anesthesia with Sevoflurane. Dig Dis Sci 59, 1415–1427 (2014). https://doi.org/10.1007/s10620-013-3023-0

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  • DOI: https://doi.org/10.1007/s10620-013-3023-0

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