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The total triterpenes in edible mushrooms have high medicinal value, and the sclerotium has various biological activities, such as the regulation of blood pressure and blood glucose. In this study, the total triterpenes of the Pleurotus tuber-regium (Fr.) Sing Sclerotium (PTRSS) were extracted, and their hypolipidemic effects were also investigated. The infrared spectra showed that the total triterpenes were consistent with the characteristic structures of the total triterpenes before and after purification. The binding abilities of total triterpenes to sodium glycocholate, sodium taurocholate, and sodium cholate were investigated, and all of them had a good binding ability to cholate. In vivo experiments showed that zebrafish tolerated the total triterpenes from the mushroom nuclei at a maximum concentration of 500 µg/mL. A correlation analysis showed that the total triterpenes from the mushroom nuclei reduced the lipid accumulation in zebrafish induced by a high-fat diet, and the lipid-lowering effect showed a correlation with dose. [ABSTRACT FROM AUTHOR]