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Enteromorpha prolifera polysaccharide–zinc (EP–Zn), a kind of polysaccharide–zinc complex, has been shown to improve the immune response and reduce the inflammatory factors in weaned piglets. Yet, the molecular mechanism remains unclear. The present study was conducted to investigate the immunomodulating activity and anti-inflammatory mechanism of EP–Zn in mice. Different doses (350 mg kg−1, 700 mg kg−1, 1050 mg kg−1 and 1400 mg kg−1) of EP–Zn were administered to C57BL/6J mice for 28 days. The results showed that under physiological conditions, 350 mg kg−1 EP–Zn stimulated cytokine (TNF-α, IL-1β, IL-6 and IL-10) secrection, regulated the intestinal microbiota, and reduced the levels of short-chain fatty acids (SCFAs) (acetic acid and propionic acid). In addition, in the LPS-induced inflammation model, EP–Zn pretreatment effectively alleviated LPS-induced shortening of colonic length …