Induction of murine TNBS colitis is strictly controlled by a modified method using continuous inhalation anesthesia with sevoflurane.
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.


OBJECTIVE
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.
First Department of Medicine, Hamamatsu University School of Medicine, Hamamatsu, 431-3192, Japan.
Tomohiro
T
Terai
Satoshi
S
Osawa
Shinya
S
Tani
Shinji
S
Oishi
Yoshifumi
Y
Arai
Takanori
T
Yamada
Mitsushige
M
Sugimoto
Takahisa
T
Furuta
Shigeru
S
Kanaoka
Hiroaki
H
Miyajima
Ken
K
Sugimoto
D000769:Anesthesia, Inhalation / Q000379:methods*; D000777:Anesthetics / Q000008:administration & dosage; D018685:Anesthetics, Inhalation / Q000008:administration & dosage*; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D003092:Colitis / Q000139:chemically induced* / Q000378:metabolism / Q000473:pathology; D016207:Cytokines / Q000378:metabolism; D004195:Disease Models, Animal; D004334:Drug Administration Schedule; D004733:Enema; D000431:Ethanol / Q000008:administration & dosage / Q000031:analogs & derivatives; D005260:Female; D007274:Injections, Intraperitoneal; D008738:Methyl Ethers / Q000008:administration & dosage*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D060888:Real-Time Polymerase Chain Reaction; D000077149:Sevoflurane; D014302:Trinitrobenzenesulfonic Acid / Q000008:administration & dosage*
D000777:Anesthetics; D018685:Anesthetics, Inhalation; D015415:Biomarkers; D016207:Cytokines; D008738:Methyl Ethers; C062527:tribromoethanol; D000077149:Sevoflurane; D000431:Ethanol; D014302:Trinitrobenzenesulfonic Acid