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

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

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Main Authors: LIN Qiyun, HOU Pengfei, LANG Hedong
Format: Article
Language:zho
Published: Editorial Office of Journal of Army Medical University 2024-01-01
Series:陆军军医大学学报
Subjects:
Online Access:http://aammt.tmmu.edu.cn/html/202309077.htm
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author LIN Qiyun
HOU Pengfei
LANG Hedong
author_facet LIN Qiyun
HOU Pengfei
LANG Hedong
author_sort LIN Qiyun
collection DOAJ
description Objective To identify the effect of acute stress induced by composite factors on intestinal barrier dysfunction and explore the relevant mechanism. Methods 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. Results 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). Conclusion Acute stress of complex factors significantly induces intestinal injury and barrier dysfunction.
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spelling doaj.art-fb7bc5d08b5748cc9d1d81a85801de462024-01-15T05:04:36ZzhoEditorial Office of Journal of Army Medical University陆军军医大学学报2097-09272024-01-01461738310.16016/j.2097-0927.202309077Effect and mechanism of acute stress induced by composite factors on intestinal barrier dysfunctionLIN Qiyun0HOU Pengfei1LANG Hedong2Research Center for Nutrition and Food Safety, Chongqing Key Laboratory of Nutrition and Health, Institute of Military Preventive Medicine, Faculty of Military Preventive Medicine, Army Medical University (Third Military Medical University), Chongqing, 400038, ChinaResearch Center for Nutrition and Food Safety, Chongqing Key Laboratory of Nutrition and Health, Institute of Military Preventive Medicine, Faculty of Military Preventive Medicine, Army Medical University (Third Military Medical University), Chongqing, 400038, ChinaResearch Center for Nutrition and Food Safety, Chongqing Key Laboratory of Nutrition and Health, Institute of Military Preventive Medicine, Faculty of Military Preventive Medicine, Army Medical University (Third Military Medical University), Chongqing, 400038, ChinaObjective To identify the effect of acute stress induced by composite factors on intestinal barrier dysfunction and explore the relevant mechanism. Methods 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. Results 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). Conclusion Acute stress of complex factors significantly induces intestinal injury and barrier dysfunction. http://aammt.tmmu.edu.cn/html/202309077.htmcomposite factorsacute stressintestinal barrier dysfunctionmechanical barrierchemical barriermicrobial barrier
spellingShingle LIN Qiyun
HOU Pengfei
LANG Hedong
Effect and mechanism of acute stress induced by composite factors on intestinal barrier dysfunction
陆军军医大学学报
composite factors
acute stress
intestinal barrier dysfunction
mechanical barrier
chemical barrier
microbial barrier
title Effect and mechanism of acute stress induced by composite factors on intestinal barrier dysfunction
title_full Effect and mechanism of acute stress induced by composite factors on intestinal barrier dysfunction
title_fullStr Effect and mechanism of acute stress induced by composite factors on intestinal barrier dysfunction
title_full_unstemmed Effect and mechanism of acute stress induced by composite factors on intestinal barrier dysfunction
title_short Effect and mechanism of acute stress induced by composite factors on intestinal barrier dysfunction
title_sort effect and mechanism of acute stress induced by composite factors on intestinal barrier dysfunction
topic composite factors
acute stress
intestinal barrier dysfunction
mechanical barrier
chemical barrier
microbial barrier
url http://aammt.tmmu.edu.cn/html/202309077.htm
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AT langhedong effectandmechanismofacutestressinducedbycompositefactorsonintestinalbarrierdysfunction