Intestinal Immune Injury Evaluation

Parts of the intestinal tissues were washed with PBS, 0.1 g of the sample tissue was weighed and homogenized with 0.9 mL ice-cold PBS in a glass homogenizer, and then the mixture was centrifuged (3000 rpm, 20 min) to obtain the supernatant. To evaluate the extent of immune injury in the intestine, the supernatants were used to determine the concentrations of secretory IgA (sIgA), IL-1β, IL-2, IL-4,TNF-α, and IFN-ϒ, and the evaluations were performed using a commercial rat ELISA kit (Jingmei Biological Technology, Jiangsu, China), respectively, and then the concentrations of diamine oxidase (DAO) and D-Lactic acid (D-LA) were determined in the blood serum using commercial ELISA kits (Jingmei Biological Technology, Jiangsu, China) to assess the degrees of gut mucosal barrier damage according to the manufacturer’s instructions. sIgA was measured as μg/ml and cytokines pg/mL. The level of sensitivity of each kit was 0.1μg/mL for sIgA and 0.1 pg/mL for each cytokine.