AbstractThe gut microbiota plays a crucial role in regulating and maintaining the epithelial barrier, particularly during early life. Notably, patients with chronic intestinal inflammation have a dysregulated process of renewal and replenishment of the intestinal epithelial cell (IEC) barrier, which is linked to disturbances in the gut microbiota. To date, there are no studies focussed on understanding the impact of inflammatory cell shedding events during the early life developmental window, and which host and microbial factors mediate these responses. Here we sought to determine pathological cell shedding outcomes throughout the postnatal developmental period (day 14, 21, 29 and week 8). Surprisingly neonatal mice (day 14 and 21) were highly refractory to induction of cell shedding after intraperitoneal administration of LPS, with day 29 mice showing strong pathological responses, more similar to those observed in adult mice. These differential responses were not linked to defects in the cellular mechanisms and pathways known to regulate cell shedding responses, although we did observe that neonatal mice had elevated anti-inflammatory (IL-10) responses. Notably, when we profiled microbiota and metabolites from these mice, we observed significant alterations. Neonatal mice had high relative abundances of Streptococcus, Escherichia and Enterococcus and increased primary bile acids. In contrast, older mice were dominated by Candidatus Arthromitus, Alistipes and Lachnoclostridium, and had increased concentrations of SCFAs and methyamines. Faecal microbiota transplant (FMT) and antibiotic studies confirmed the importance of early life gut microbiota in cell shedding responses. In these studies, neonates treated with antibiotics restored LPS-induced small intestinal cell shedding, whereas adult FMT alone had no effect. Our findings further support the importance of the early life window for microbiota-epithelial interactions in the presence of inflammatory stimuli and highlight areas for further investigation to probe underlying mechanisms to drive therapeutic development within the context of chronic inflammatory intestinal diseases.

An increased production of free radicals: 0.105 AE 0.023 vs. 0.073 AE 0.022 mM/min/mg (p < 0.001).In the preconditioning group, mitochondrial respiration is restored in IL (Vmax ¼ 8.85 AE 2.07 vs. 9.08 AE 1.89 mmol02/min/g), and production of free radicals is reduced (0.084 AE 0.018 vs. 0.082 AE 0.027 mM/min/mg). There is no difference between IL and CL. In the post-conditioning group, there is a significant alteration, both in IL and CL, with impaired mitochondrial respiration (Vmax ¼ 5.03 AE 1.14 vs. 6.78 AE 1.29 mmol02/min/g) and increased production of free radicals (0.161 AE 0.077 vs. 0.092 AE 0.052 mM/min/mg).Conclusion: Statins have protective effects on skeletal critical ischemic muscle in primary prevention. They have deleterious effects in secondary prevention, by alteration of the mitochondrial respiratory function and by increased production of free radicals.

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