Psoralen prevents the inactivation of estradiol and treats osteoporosis via covalently targeting HSD17B2. (15th July 2023)

• Record Type: Journal Article
• Authors: Lu, Yujie; Zhang, Man; Zhang, Jin; Jiang, Min; Bai, Gang
• Is Part Of: Journal of ethnopharmacology. Volume 311(2023)
• Publication Date: 2023-07-15
• Subjects: Psoralen -- Covalent binding -- HSD17B2 -- Anti-Osteoporosis -- Estradiol; Pso psoralen -- aPso alkynyl-modified Pso probe -- HSD17B2 17-β hydroxysteroid dehydrogenase type 2 -- E2 estradiol -- OBs osteoblasts -- OCs osteoclasts -- NAD+ nicotinamide-adenine dinucleotide -- CETSA cellular thermal shift assays -- BDP TMR boron-dipyrromethene tetramethylrhodamine -- NH2-MMs Fe3O4 amino magnetic microspheres -- N3-MMs azide-modified magnetic microspheres -- OVX ovariectomized -- ALP alkaline phosphatase -- PCC Pearson correlation coefficient -- BMD bone mineral density -- BMC bone mineral content; Ethnopharmacology -- Periodicals; Pharmacognosy -- Periodicals; Herbs -- Periodicals; Herbs -- Periodicals; Pharmacognosy -- Periodicals; Pharmacognosie -- Périodiques; Herbes -- Périodiques; 615.1
• DOI: 10.1016/j.jep.2023.116426
• Languages: English
• ISSNs: 0378-8741

Abstract

Abstract: Ethnopharmacological relevance: Psoralea corylifolia L. seeds ( P. corylifolia ), popularly known as Buguzhi in traditional Chinese medicine, are often used to treat osteoporosis in China. Psoralen (Pso) is the key anti-osteoporosis constituent in P. corylifolia, however, its targets and mechanism of action are still unclear. Aim of the study: The purpose of this study was to explore the interaction between Pso and 17-β hydroxysteroid dehydrogenase type 2 (HSD17B2), an estrogen synthesis-related protein that inhibits the inactivation of estradiol (E2) to treat osteoporosis. Materials and methods: Tissue distribution of Pso was analyzed by in-gel imaging after oral administration of an alkynyl-modified Pso probe (aPso) in mice. The target of Pso in the liver was identified and analyzed using chemical proteomics. Co-localization and cellular thermal shift assays (CETSA) were used to verify the key action targets. To detect the key pharmacophore of Pso, the interaction of Pso and its structural analogs with HSD17B2 was investigated by CETSA, HSD17B2 activity assay, and in-gel imaging determination. Target competitive test, virtual docking, mutated HSD17B2 activity, and CETSA assay were used to identify the binding site of Pso with HSD17B2. A mouse model of osteoporosis was established by ovariectomies, and the efficacy of Pso in vivo was confirmed by micro-CT, H&E staining, HSD17B2 activity, and bone-related biochemical assays. Results: Pso regulated estrogen metabolism by targeting HSD17B2 in the liver, with the α, β-unsaturated ester in Pso being the key pharmacophore. Pso significantly suppressed HSD17B2 activity by irreversibly binding to Lys236 of HSD17B2 and preventing NAD + from entering the binding pocket. In vivo studies in ovariectomized mice revealed that Pso could inhibit HSD17B2 activity, prevent the inactivation of E2, increase levels of endogenous estrogen, improve bone metabolism-related indices, and play a role in anti-osteoporosis. Conclusions: Pso covalently binds to Lys236 of HSD17B2 in hepatocytes to prevent the inactivation of E2, thereby aiding in the treatment of osteoporosis. Graphical abstract: Image 1 Highlights: Psoralen targets HSD17B2 regulating estrogen synthesis in liver. The α, β-unsaturated ester of psoralen covalently binds with Lys236 to inhibit HSD17B activity. Psoralen prevents the inactivation of estradiol in vivo to treat osteoporosis.