Key factors in developing the trinitrobenzene sulfonic acid-induced post-inflammatory irritable bowel syndrome model in rats

Table 5 Detailed protocols in developing trinitrobenzene sulfonic acid induced post-inflammatory irritable bowel syndrome model in Sprague-Dawley rats

Procedures	Cautions
1. 7-wk-old male Sprague-Dawley rats, fast 24 h before TNBS administration
2. Mix 1 volume of 12.5 mg/mL TNBS-saline solution with 1 volume of absolute ethanol	Fresh preparation
3. Weight and anesthetized rats with chloral hydrate (350 mg/kg, i.p.)	Anesthetization may last for 3-4 h
4. Insert a catheter into the colon at 8 cm from anus	Proceed carefully to avoid damage the colon wall
5. Keep the rat with head-down vertical position, instill 0.8 mL TNBS solution slowly into the colon lumen within 1 min	Handle slowly to avoid TNBS leakage
6. Keep the rat in head-down vertical position for 1 min before gently removing the catheter
7. Put the rats in a mound of bedding in head-down position until consciousness recovery	Ensure TNBS solution remains completely in the colon; Keep the rats warm
8. On day 3, 7, 14, 21 and 28, observe and weight the rats	On day 3, body weight decreased by 10%-20% with unformed bloody stool;
	On day 7, body weight regained and reached the original level, the stool become formed but still soft;
	From day 7 to day 28, body weight increased smoothly, and finally reached the control level

TNBS: Trinitrobenzene sulfonic acid.