Ever-increasingly Citrus wastes have been generating during the industrial processing, which is troublesome to dispose them for the considerations of economic feasibility and environmental protection. Meanwhile, liver disease, which causes liver damage, is the one of the major threats for the further development of aquaculture, especially in marine fish. The present study explored the hepatoprotective effect of PSCP, a polysaccharide extracted from the pomelo seed coat, in the primary hepatocytes of hybrid grouper (<i>Epinephelus fuscoguttatus</i>♂ × <i>Epinephelus lanceolatus</i>♀). PSCP displayed considerably scavenging effect to the free radicals and strong inhibitive effect to the erythrocyte hemolysis. In vitro, we added PSCP (0, 100, 200 and 400 μg/mL) to the primary hepatocytes before incubation with Acetaminophen (APAP) (12 mM). The CKK-8 experiment displayed the administration of PSCP (100 and 200 μg/mL) substantially mitigated the reduction of hepatocyte viability inflicted by APAP. Moreover, we found that PSCP effectively alleviated APAP-induced oxidative stress, as shown by the significantly reduced level of reactive oxygen species and the elevated enzymatic activities of superoxide dismutase, catalase, and glutathione peroxidase, which indicated its anti-liver injury effect (P<0.05). The more direct signs associating with the hepatoprotection of PSCP are reflected in the results of hematoxylin and eosin staining, as evident by the morphological recovery following the addition of PSCP. In brief, these findings showed that the therapeutic potential of PSCP on APAP-induced fish liver injury, which not only provided a now prospect in treating the liver impairment in aquatic animals, but to also improve the utilization of pomelo fruitlets.