Effect and mechanism of acute stress induced by composite factors on intestinal barrier dysfunction

To identify the effect of acute stress induced by composite factors on intestinal barrier dysfunction and explore the relevant mechanism.

Twenty male C57BL/6 mice (7-8 weeks old, body mass 20-22 g) were randomly divided into control group (CON group, n=10) and model group (MOD group, n=10). To establish an acute stress model of complex factors, the mice of the MOD group were given rest of 4 h, sleep deprivation of 15 h, and then followed by 3 h of noise (120 dB)/flash (2 000±500 lx) and 2 h of forced swimming (water temperature 26±1 ℃, water depth 45 cm) within 24 h. While the mice in the CON group were placed in a noiseless environment with constant temperature and humidity and normal circadian rhythm but been given no any treatment. At the end of forced swimming, the mice from the MOD group were anesthetized for blood samples and then euthanized. Serum markers of intestinal injury were detected by immunoenzyme-linked adsorption assay kit, morphological changes in colonic tissues were observed by HE staining, protein and mRNA levels of colonic tissue barrier function related molecules and inflammatory factors were detected by immunofluorescence, immunohistochemistry and RT-qPCR, and intestinal flora changes were analyzed by 16S rRNA technique.

Compared with the CON group, the serum markers of intestinal damage, lipopolysaccharide (LPS), intestinal fatty acid binding protein (I-FABP) and Zonulin were significantly increased in the MOD group (P < 0.05). HE staining showed that intestinal tissue damage was obvious in the MOD group. The results of immunofluorescence staining and RT-qPCR showed that the levels of mechanical barrier related molecules Zona occludens 1 (ZO-1) and Occludin were obviously decreased (P < 0.05). Immunohistochemistry and PAS staining indicated the number of chemical barriers associated goblet cells, and expression of mucin2 (Muc-2), antimicrobial peptide regenerating islet derived-3γ (Reg3γ) and regenerating islet derived-3β (Reg3β) were notably reduced (P < 0.05). Intestinal flora β diversity significantly changed, with harmful bacteria Bacteroidetes and Desulfovibrio significantly increased, while beneficial bacteria Rumenococcus family, norank_f_Erysipelotrichaceae, Gemella genus and Erysipelotrichaceae genus declined (P < 0.05). The levels of TNF-α, IL-6 and ROS were significantly enhanced (P < 0.05).

Acute stress of complex factors significantly induces intestinal injury and barrier dysfunction.