Increased beverage consumption has long been recognized to play an important role in the pathogenesis of metabolic diseases and colorectal cancer (24–26). Although food additives such as emulsifiers, food colorants, titanium dioxide, and aluminum have largely contributed to the development of these conditions, recent evidence have also identified that food additives are pathogenic factors for colitis (26, 27). In this study, we focused on the potential contribution of cold stress associated with beverage consumption to IBD development. Our results revealed that cold stress was a novel risk factor for colitis in mice, including induction of gut barrier injury, increased anxiety-like behaviors, gut microbiota disorder, and metabolite disturbance. These findings suggest that exposure to cold beverages is also a novel risk factor for IBD in humans and a potential trigger for establishing experimental IBD in mice.
Dysregulation of the gut barrier in IBD is caused by environmental factors and genetic predisposition (28), however, identifying specific environmental factors has been difficult. Several food additives are identified as environmental risk factors for IBD (27, 29). Here, we aimed to identify whether the beverage temperature directly affect the development of colitis. Similar to observations in humans, the administration of cold water to mice inhibited gut transit, additionally, the mice exhibited a colitis-like phenotype of gut barrier injury. The fact that increased cold beverage consumption is related to some gastrointestinal symptoms seemingly suggests a link between cold stress and intestinal disorders. Indeed, our results indicate that exposure to cold water not only induces gut transit disturbances but also produces low-grade inflammation. In line with these considerations, cold water exposure expectedly led to activation of ubiquitous inflammatory signaling pathways, suggesting local mucosal immune cell activation and immune cell trafficking, which are characteristics of IBD (30).
Low-grade inflammation in colitis is associated with and may be promoted by gut microbiota dysbiosis (31). The low bacterial diversity and altered gut microbiota observed in the cold stress group in this study are extremely similar to those observed in IBD. For example, microbial diversity studies have demonstrated the overgrowth of Proteobacteria in IBD patients (32). Under normal homeostasis, epithelial cells, tight junctions, and the local immune system prevent the translocation of pathogens in the gut. However, in genetically susceptible individuals, Proteobacteria expand to colonize the lumen and invade the lamina propria, further aggravating disruption of the gut barrier (33). The host recognizes Proteobacteria via nucleotide oligomerization domain-like receptors, Toll-like receptors and retinoic acid-inducible gene I-like receptors. Moreover, microbiota-derived products such as lipopolysaccharide, peptidoglycan, and flagellin from Proteobacteria trigger the activation of immune responses in the mucosa. Our study suggests that the increased abundance of pathogens accompanied by cold stress may contribute to gut barrier disruption in mice by accelerating inflammation in the gut.
Apart from the microbiota-derived products, the metabolites of the gut microbiota are also key actors in the development and exacerbation of IBD. Accumulating evidence suggests that signals from microbial metabolites affect mucosal integrity and immune homeostasis. Moreover, in IBD patients, the metabolites composition and function are disturbed seriously, including bile acids, short-chain fatty acids and tryptophan (34, 35). Significant alteration in the gut microbiota metabolites was also identified under cold stress exposure in this study. Interestingly, we found a remarkable reduction in the levels of several metabolites in the dopamine-related pathway.
Dopamine is an critical catecholaminergic neurotransmitter that present in the peripheral tissues and central nervous system simultaneously, which regulates blood pressure, sodium balance, glucose homeostasis, cognition, memory, the sympathetic nervous system, and mood (36). Dopamine is mainly synthesized in the brain, T cells, dendritic cells, and as well as by gut commensals such as members of the genus Clostridium (36). Dopamine has recently recognized as an important regulator of the immune system. Disturbance of the dopamine pathway affects both innate and adaptive immunity largely, causing the development of inflammatory pathologies (37). A significant proportion of patients with gut barrier injury suffer from anxiety and depression (38), implying a rational connection between the gut and the brain. Consistently, we examined their behaviors and found that cold water lead to a tendency to depression. Our findings suggest that cold water stress result in reduced neuro-activities in gut microbiota and enhanced pro-inflammatory activity that promotes anxiety and depression.
Nevertheless, our study has several limitations. We handled mice with cold water to explore the effect of cold stress to the gut barrier, microbiota, and metabolites. These findings acknowledge the important role of cold stress, however, it cannot mimic the condition of cold beverage consumption completely in humans because of the variety of beverages used and the differences between human and mouse physiology. In addition, although we find a significant overgrowth of pathogens and decreased dopamine-related metabolites in cold water-treated mice, we cannot completely identify that the alteration in gut microbiota causes the gut barrier injury. Further studies to investigate the role of microbiota and their metabolites with the effect on gut barrier and behavior are also needed to deeply elucidate their causal or accompanied relationships.