Characterization of PCS-2A, a polysaccharide derived from chestnut shell, and its protective effects against HO-induced liver injury in hybrid grouper.

PCS-2A is a 34,023-Da acidic polysaccharide purified from chestnut shell consisting of rhamnose, arabinose, galactose, glucose, ribose, and galacturonic acid subunits at a molar ratio of 0.019:0.044:0.059:0.052:0.197:0.628. FTIR, methylation, and NMR analyses suggest the following backbone, (→4)-α-d-GalAp-(1 → 2,4)-α-l-Rha-(1→), with the branch chain composed of arabinose on O-2 with 2,4)-α-l-Rha-(1→). CCK-8 assay indicated PCS-2A treatment offset the reduction in cell viability inflicted by HO. Furthermore, histological signs of recovery in hepatocytes and liver tissue and a decreased level of AST and ALT occurred following administration of PCS-2A, indicating anti-liver lesion capability. In addition, we found that PCS-2A effectively alleviated HO-induced oxidative stress via activation of the NRF2 signaling pathway, evidenced by the downregulation of ROS content and upregulation of Nrf2 expression, as well as its corresponding antioxidant enzymes. The antioxidative effect elicited by PCS-2A further ameliorated NF-κB-mediated inflammation, as evidenced by lower mRNA levels of inflammatory cytokines, higher IκB in vitro, and reduced gene expression and activities of proinflammatory cytokines in vivo. Furthermore, in vitro apoptosis-related indicators revealed that P53-mediated apoptosis was alleviated via oxidative stress modulation. In summary, these results suggest that PCS-2A may elicit a protective effect against HO-induced liver injury via upregulation of the NRF2 signaling pathway.