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Abstract
Growing evidence has demonstrated that stress triggers gastrointestinal (GI) disorders. This study aimed to investigate how the acute cold water-immersion restraint (CWIR) stress affects intestinal injury and gut microbiota (GM) distribution. Male C57BL/6 mice were used to establish a CWIR animal model. Hematoxylin–eosin and periodic acid–Schiff staining were performed to assess intestinal histopathological changes. Reverse transcription quantitative polymerase chain reaction (RT-qPCR) analysis and immunofluorescence staining were used to evaluate the expression of inflammatory cytokines and immune cell infiltration in the intestinal tissues. The gut permeability and intestinal occludin protein expression were determined through fluorescein isothiocyanate-dextran detection and western blot, respectively. GM profiles were analyzed via high-throughput sequencing of the fecal bacterial 16S rRNA genes. Results showed that CWIR induced more severe intestinal mucosal injury compared to the control, leading to a significant increase in tumor necrosis factor-α expression, but no infiltration of neutrophil and T cells. CWIR also resulted in GI disruption and increased the permeability of the intestinal mucosa. GM profiles showed that CWIR reduced GM diversity of mice compared with the control group. Specifically, aerobic and gram-negative bacteria significantly increased after CWIR, which was associated with the severity of gut injury under stress. Therefore, acute CWIR leads to severe intestinal damage with inflammation and disrupts the GM homeostasis, contributing to decreased GM diversity. Our findings provide the theoretical basis for the further treatment of intestinal disorders induced by CWIR.
Highlights
Stress can be caused by various external or internal stimuli, leading to the emergence of strong defense systems, which may have a beneficial or harmful impact on the body (Dragos and Tanasescu, 2010; Yaribeygi et al, 2017)
The intestinal mucosa in the control group exhibited a complete structure, which was disrupted by cold water-immersion restraint (CWIR) treatment (Figure 1A)
Mice subjected to CWIR exhibited significantly reduced goblet cell numbers in all the intestinal segments (Figure 2B)
Summary
Stress can be caused by various external or internal stimuli, leading to the emergence of strong defense systems, which may have a beneficial or harmful impact on the body (Dragos and Tanasescu, 2010; Yaribeygi et al, 2017). Acute stresses like cold-restraint, restraint, or waterimmersion restraint can simulate the pathophysiology of stress-related mucosal disease (SRMD), Acute CWIR Reduces GM Diversity highly prevalent in patients in intensive care units. Among these three stress models, water-immersion restraint best resembles SRMD, as previously shown by the significantly elevated ulcer index, microvascular permeability, and decreased hexosamine level (Saxena and Singh, 2017). Ohta et al demonstrated that WIRS exposure of rats induced severe oxidative stress in immune organs (Ohta et al, 2012). The pathogenesis of intestinal mucosal injury induced by water restraint stress remains unclear
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