Evaluation of Intestinal Morphology

To investigate intestinal morphology, paraformaldehyde-fixed jejunum, ileum, and duodenum were dehydrated and embedded in paraffin. Next, 4-µm sections were cut and then stained with hematoxylin and eosin stain. Intestinal morphology was carried out with a light microscope (Leica, Solms, Germany) with the Leica Application Suite image analysis software (Leica, Solms, Germany) [14 (link)]. There are 6 villus and crypts that were counted per histological cutting. Intestinal villus height, crypt depth, and villus surface area were measured to calculate the ratio of villus height to crypt depth. The activities of GSH-Px, SOD, and CAT and the contents of MDA and H₂O₂ were determined by using commercially available kits (Jiancheng Bioengineering Institute, Nanjing, China).