Menozzi Alessandro, Ossiprandi Maria Cristina
University of Parma, Parma, Italy.
Curr Protoc Toxicol. 2010 May;Chapter 21:Unit 21.3. doi: 10.1002/0471140856.tx2103s44.
The disruption of intestinal barrier leads to the penetration of noxious luminal compounds into the gut wall, causing further damage. This unit describes the assessment of enteric bacteria translocation into the intestinal wall of rats, an established method for the evaluation of bowel damage to the mucosal epithelial barrier. The Basic Protocol provided in the present unit describes collection and preparation of small intestine sample, performing of sample serial dilutions for bacterial culture, performing of the culture of aerobic and anaerobic bacteria on petri dishes, incubation of the cultured plates, and counting of bacterial colonies. The Support Protocols describes the procedures for the preparation of petri dishes for the culture, using different employable media for aerobes or anaerobes. The Alternate Protocol describes the use of the "inclusion method," suitable for the culture of anaerobic bacteria.
肠道屏障的破坏会导致有害的肠腔化合物渗入肠壁,进而造成进一步损伤。本单元介绍了评估肠道细菌向大鼠肠壁转移的方法,这是一种评估肠道黏膜上皮屏障损伤的既定方法。本单元提供的基本方案描述了小肠样本的采集和制备、进行细菌培养的样本系列稀释、在培养皿上进行需氧菌和厌氧菌培养、培养平板的孵育以及细菌菌落计数。支持方案描述了用于培养的培养皿的制备程序,使用适用于需氧菌或厌氧菌的不同可用培养基。替代方案描述了适用于厌氧菌培养的“包埋法”的使用。