Obeticholic acid and INT-767 modulate collagen deposition in a NASH in vitro model

Beatrice Anfuso,Claudio Tiribelli,Natalia Rosso,Luciano Adorini

doi:10.1038/s41598-020-58562-x

Beatrice Anfuso, Claudio Tiribelli + Show 2 more

Open Access

https://doi.org/10.1038/s41598-020-58562-x

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Abstract

Pharmacological treatments for non-alcoholic steatohepatitis (NASH) are still unsatisfactory. Fibrosis is the most significant predictor of mortality and many anti-fibrotic agents are under evaluation. Herein, we assessed in vitro the effects of the FXR agonist obeticholic acid (OCA) and the dual FXR/TGR5 agonist INT-767 in a well-established co-culture NASH model. Co-cultures of human hepatoma and hepatic stellate (HSCs) cells were exposed to free fatty acids (FFAs) alone or in combination with OCA or INT-767. mRNA expression of HSCs activation markers and FXR engagement were evaluated at 24, 96 and 144 hours. Collagen deposition and metalloproteinase 2 and 9 (MMP2-9) activity were compared to tropifexor and selonsertib. FFAs induced collagen deposition and MMP2-9 activity reduction. Co-treatment with OCA or INT-767 did not affect ACTA2 and COL1A1 expression, but significantly reduced FXR and induced SHP expression, as expected. OCA induced a dose-dependent reduction of collagen and induced MMP2-9 activity. Similarly, INT-767 induced collagen reduction at 96 h and a slight increase in MMP2-9. Tropifexor and Selonsertib were also effective in collagen reduction but showed no modulation of MMP2-9. All tested compounds reduced collagen deposition. OCA exerted a more potent and long-lasting effect, mainly related to modulation of collagen turn-over and MMP2-9 activity.

Highlights

Pharmacological treatments for non-alcoholic steatohepatitis (NASH) are still unsatisfactory
Obesity prevalence is booming in both hig- and low-income countries has led to a surge in non-alcoholic fatty liver disease (NAFLD), a condition characterized by liver steatosis
Several models describing NASH pathogenesis propose that when the amount of free fatty acids (FFAs) in hepatocytes is overwhelmed, lipotoxic species formation leads to hepatocyte injury, inflammatory cells recruitment, and hepatic stellate cells (HSCs) activation

Summary

Introduction

Pharmacological treatments for non-alcoholic steatohepatitis (NASH) are still unsatisfactory. FFAs induced collagen deposition and MMP2-9 activity reduction. OCA exerted a more potent and long-lasting effect, mainly related to modulation of collagen turn-over and MMP2-9 activity. Several models describing NASH pathogenesis propose that when the amount of free fatty acids (FFAs) in hepatocytes is overwhelmed, lipotoxic species formation leads to hepatocyte injury, inflammatory cells recruitment, and hepatic stellate cells (HSCs) activation. FXR is down-regulated in patients with NAFLD and its activation exerts beneficial effects counteracting obesity, steatosis, inflammation, and fibrosis[5]. Due to these favorable effects in chronic liver disease, several small molecules activating FXR are being developed[7,8]

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