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

Abstract

Background: Activated 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).

Aim: To elucidate the mechanism of the anti-fibrotic effect of DX and determine whether it induces senescence.

Methods: Primary 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.

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. PMA and m-3M3FBS reversed the effect of DX on senescence markers.

Conclusion: Doxazosin reverses the fibrogenic phenotype of aHSCs and induces the senescence phenotype.

Keywords: Cell proliferation; Liver fibrosis; Norepinephrine; Phospholipase C; Protein kinase C; Senescence; Stellate cells; α1-Adrenergic signaling.