Python-derived 16α-Hydroxylated Bile Acid, Pythocholic Acid is a ligand for TGR5, not farnesoid X receptors and vitamin D receptors

Bile acids (BAs) are signaling molecules involved in energy expenditure, glucose homeostasis, and immune system regulation via activation of BA receptors, such as Takeda G-Protein-Coupled Receptor 5 (TGR5), Farnesoid X Receptor (FXR), and Vitamin D Receptor (VDR). The structure of BA, especially the hydroxyl group position, plays an important role in exerting its function. Previously, we reported that 16α-hydroxylated BA, also known as pythocholic acid (PCA), has beneficial effects on metabolic function and lipid metabolism in mammals. However, the molecular mechanism of PCA in mammals is yet to be explored because 16α-hydroxylated BA has not been seen in mammals. This study aims to investigate the binding interaction of PCA to human bile acid receptors, TGR5, FXR, and VDR, using a luciferase reporter assay. Luciferase reporter assay showed that PCA and tauro-conjugated-PCA (TPCA) activated TGR5, but did not activate FXR or VDR. Additionally, PCA and TPCA did not show an antagonistic effect on any of the BA receptors. TPCA treatment significantly decreased lipopolysaccharide (LPS)-induced tumor necrosis factor-alpha (TNF-α) expression in mouse peritoneal macrophages, and inhibition of TGR5 by SBI-115 canceled the anti-inflammatory effect of TPCA. Our data suggests that PCA and TPCA are ligands for mammalian TGR5 receptors. •Pythocholic acid and its tauro conjugation were identified as a human TGR5 agonist.•Pythocholic acid does not activate FXR and vitamin D receptors.•Tauro-conjugated pythocholic acid suppresses inflammatory response in macrophages.