Protective Effects of Squid Gonad Phospholipids Against Hepatotoxicity Induced by Multi‐Glycoside From Tripterygium wilfordii (GTW): Inhibition of Inflammatory Response and Boost of GTW Metabolism in Zebrafish

ABSTRACT Multi‐glycoside from Tripterygium wilfordii Hook.f. (GTW) has shown clinical efficacy in suppressing immunological dysfunction, but is associated with severe hepatotoxicity. Phospholipids derived from marine organisms, particularly enriched in long‐chain polyunsaturated fatty acids, possess various beneficial bioactivities and significant medicinal value. However, the antagonistic action of phospholipids against GTW‐induced hepatotoxicity remains unclear. In this study, we explored the protective effect of squid gonads–derived phospholipids (SPLs) against GTW‐induced hepatotoxicity using a zebrafish model. We evaluated the changes in a range of parameters following SPLs co‐treatment, including liver morphology, histology, transaminase activities, microRNA‐122 (miR‐122) transcript level, and expressions of genes related to liver injury. The results indicated that SPLs significantly attenuated GTW‐induced malformations and changes in the liver and yolk sac areas of zebrafish. The abnormal elevation of transaminase activities and dramatic downregulation of miR‐122 induced by GTW were remarkably reversed by SPLs. Histological and ultrastructural examinations showed SPLs alleviated GTW‐induced abnormalities in the liver tissue, including loose cell‐to‐cell contact, vacuolar structures, focal necrosis, and mitochondrial ultrastructural abnormalities. Moreover, SPLs significantly reversed the abnormal expressions of genes related to inflammation within the Myd88/NF‐κB signaling pathway, and those involved in drug/xenobiotic metabolism. Molecular docking simulations revealed that these key inflammatory and drug metabolic regulators exhibited stable docking affinities with seven major components of SPLs. These findings suggest that SPLs may exert protective effects against GTW‐induced hepatotoxicity, potentially through the inhibition of inflammatory responses and the restoration of GTW metabolism. Therefore, SPLs may serve as a promising therapeutic candidate for mitigating GTW‐induced hepatotoxicity.