Scope: Hepatic encephalopathy (HE) refers to neurological dysfunction associated with hepatic inadequacy and gut dysbiosis. Chitooligosaccharides (COS) possesses prominent biological activities including incalculable hepatoprotective, neuroprotective and prebiotic effects. This study evaluates the protective effects of COS on HE from the influence of gut-liver-brain axis in mice. Methods and results: Hepatic injured mice show minimal symptoms of HE, reflecting in cognitive impairment, and learning and memory retardation, while they are reversed by COS following orally administrated. Furthermore, COS ameliorates brain function through inhibiting microglial and astrocyte activation in cerebral cortex and hippocampus, promoting neuronal regeneration characterized by the increase of neuron-specific marker (neuronal nuclear antigen, NeuN). Concurrently, neuroinflammation and hepatitis are restrained by COS through descending toll-like receptors 4/Nuclear factor kappa B (TLR4/NF-𝜿B) pathway. Additionally, the dysbiosis of the composition and structure of gut microbiota is displayed in mice with HE, while it is modified by COS through decreasing the relative abundances of Muribaculaceae, Lactobacillus, and Enterorhabdus. The enhancement of blood ammonia is crucially slipped to basal levels by COS. Conclusion: The present study shows that COS could prevent the pathological process of HE through regulating the gut-liver-brain cross-talk, which provids new insight into fundamental roles of COS.