Highlights
- •Mebhydrolin as a selective FXR antagonist improves glucose homeostasis in T2DM mice.
- •Mebhydrolin regulated both gluconeogenesis and glycogen synthesis in T2DM mice.
- •Mebhydrolin suppresses gluconeogenesis through FXR/miR-22-3p/PI3K/AKT/FoxO1 pathway.
- •Mebhydrolin promotes glycogen synthesis through FXR/miR-22-3p/PI3K/AKT/GSK3β pathway.
- •Mebhydrolin binds to residues L291, M332 and Y373 of FXR-LBD.
Abstract
Introduction
Type 2 diabetes mellitus (T2DM) is a chronic disease with hallmarks of hyperglycemia
and hyperlipidemia. Long-term hyperglycemia damages the functions of multiple tissues
and organs leading to a series of complications and disability or even death. Nuclear
receptor farnesoid X receptor (FXR) antagonism has been recently discovered to exhibit
beneficial effect on glucose metabolism in T2DM mice, although the underlying mechanisms
remain unclear. Here, we performed the study on the discovery of new FXR antagonist
and investigated the mechanism underlying the amelioration of FXR antagonism on glucose
homeostasis in T2DM mice by using the determined FXR antagonist as a probe.
Methods
FXR antagonist Mebhydrolin was discovered by screening against the lab in-house FDA
approved drug library through surface plasmon resonance (SPR), microscale thermophoresis
(MST), AlphaScreen, mammalian one-hybrid and transactivation assays. Activity of Mebhydrolin
in improving glucose homeostasis was evaluated in db/db and HFD/STZ-induced T2DM mice, and the mechanisms governing the regulation of Mebhydrolin
were investigated by assays of immunostaining, Western blot, ELISA, RT-PCR against
liver tissues of both T2DM mice and the T2DM mice with liver-specific FXR knockdown
injected via adeno-associated-virus AAV-FXR-RNAi and mouse primary hepatocytes. Finally,
molecular docking and molecular dynamics (MD) technology-based study was performed
to investigate the structural basis for the antagonistic regulation of Mebhydrolin
against FXR at an atomic level.
Findings
Mebhydrolin ameliorated blood glucose homeostasis in T2DM mice by both suppressing
hepatic gluconeogenesis via FXR/miR-22-3p/PI3K/AKT/FoxO1 pathway and promoting glycogen
synthesis through FXR/miR-22-3p/PI3K/AKT/GSK3β pathway. Structurally, residues L291,
M332 and Y373 of FXR were required for Mebhydrolin binding to FXR-LBD, and Mebhydrolin
induced H2 and H6 shifting of FXR potently affecting the regulation of the downstream
target genes.
Conclusions
Our work has revealed a novel mode for the regulation of FXR against glucose metabolism
in T2DM mice and highlighted the potential of Mebhydrolin in the treatment of T2DM.
Keywords
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Article info
Publication history
Published online: April 04, 2021
Accepted:
March 28,
2021
Received:
November 6,
2020
Identification
Copyright
© 2021 Elsevier Inc. All rights reserved.
