α-1 Adrenergic receptor antagonist doxazosin reverses hepatic stellate cells activation via induction of senescence

Sandra A Serna-Salas,Johanna C Arroyave-Ospina,Mengfan Zhang,Turtushikh Damba,Manon Buist-Homan,Martin H Muñoz-Ortega,Javier Ventura-Juárez,Han Moshage

doi:10.1016/j.mad.2021.111617

Abstract

BackgroundActivated hepatic stellate cells (aHSCs) are the main effector cells during liver fibrogenesis. α-1 adrenergic antagonist doxazosin (DX) was shown to be anti-fibrotic in an in vivo model of liver fibrosis (LF), but the mechanism remains to be elucidated. Recent studies suggest that reversion of LF can be achieved by inducing cellular senescence characterized by irreversible cell-cycle arrest and acquisition of the senescence-associated secretory phenotype (SASP). AimTo elucidate the mechanism of the anti-fibrotic effect of DX and determine whether it induces senescence. MethodsPrimary culture-activated rat HSCs were used. mRNA and protein expression were measured by qPCR and Western blot, respectively. Cell proliferation was assessed by BrdU incorporation and xCelligence analysis. TGF-β was used for maximal HSC activation. Norepinephrine (NE), PMA and m-3M3FBS were used to activate alpha-1 adrenergic signaling. ResultsExpression of Col1α1 was significantly decreased by DX (10 μmol/L) at mRNA (−30 %) and protein level (−50 %) in TGF-β treated aHSCs. DX significantly reduced aHSCs proliferation and increased expression of senescence and SASP markers. PMA and m-3M3FBS reversed the effect of DX on senescence markers. ConclusionDoxazosin reverses the fibrogenic phenotype of aHSCs and induces the senescence phenotype.

Highlights

Liver fibrosis is the excessive deposition of collagen type 1 in the extracellular matrix (ECM) as a result of an uncontrolled wound-healing response caused by chronic liver inflammation
We demonstrated that the α1-Adrenergic receptors (AR) antagonist doxazosin decreases collagen deposition in liver tissue in vivo (Serna-Salas et al, 2018), the exact anti-fibrotic mechanism of doxazosin has not been elucidated yet
We show that doxazosin targets Activated hepatic stellate cells (aHSCs), by decreasing expression of collagen type 1 at protein and mRNA

Summary

Introduction

Liver fibrosis is the excessive deposition of collagen type 1 in the extracellular matrix (ECM) as a result of an uncontrolled wound-healing response caused by chronic liver inflammation. Hepatic stellate cells undergo an activation process and transdifferentiate into myofibroblast-like cells, accompa nied by several phenotypic changes, such as de novo expression of alphasmooth muscle actin (αSMA), increased proliferation, loss of retinoidstoring capacity and the acquisition of fibrogenic functions (Rippe and Brenner, 2004). It has been described that senescence, a state of irreversible cell cycle arrest, may (partially) reverse stellate cell activation. Recent studies suggest that reversion of LF can be achieved by inducing cellular senescence characterized by irreversible cell-cycle arrest and acquisition of the senescence-associated secretory phenotype (SASP). Results: Expression of Col1α1 was significantly decreased by DX (10 μmol/L) at mRNA (− 30 %) and protein level (− 50 %) in TGF-β treated aHSCs. DX significantly reduced aHSCs proliferation and increased expression of senescence and SASP markers. Conclusion: Doxazosin reverses the fibrogenic phenotype of aHSCs and induces the senescence phenotype

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