Carvedilol Inhibits Angiotensin II-Induced Proliferation and Contraction in Hepatic Stellate Cells through the RhoA/Rho-Kinase Pathway

Biomed Res Int. 2019 Nov 7:2019:7932046. doi: 10.1155/2019/7932046. eCollection 2019.

Abstract

Aim: Carvedilol is a nonselective beta-blocker used to reduce portal hypertension. This study investigated the effects and potential mechanisms of carvedilol in angiotensin II- (Ang II-) induced hepatic stellate cell (HSC) proliferation and contraction.

Methods: The effect of carvedilol on HSC proliferation was measured by Cell Counting Kit-8 (CCK-8). Cell cycle progression and apoptosis in HSCs were determined by flow cytometry. A collagen gel assay was used to confirm HSC contraction. The extent of liver fibrosis in mice was evaluated by hematoxylin-eosin (H&E) and Sirius Red staining. Western blot analyses were performed to detect the expression of collagen I, collagen III, α-smooth muscle actin (α-SMA), Ang II type I receptor (AT1R), RhoA, Rho-kinase 2 (ROCK2), and others.

Results: The results showed that carvedilol inhibited HSC proliferation and arrested the cell cycle at the G0/G1 phase in a dose-dependent manner. Carvedilol also modulated Bcl-2 family proteins and increased apoptosis in Ang II-treated HSCs. Furthermore, carvedilol inhibited HSC contraction induced by Ang II, an effect that was associated with AT1R-mediated RhoA/ROCK2 pathway interference. In addition, carvedilol reduced α-SMA expression and collagen deposition and attenuated liver fibrosis in carbon tetrachloride (CCl4)-treated mice. The in vivo data further confirmed that carvedilol inhibited the expression of angiotensin-converting enzyme (ACE), AT1R, RhoA, and ROCK2.

Conclusions: The results indicated that carvedilol dose-dependently inhibited Ang II-induced HSC proliferation by impeding cell cycle progression, thus alleviating hepatic fibrosis. Furthermore, carvedilol could inhibit Ang II-induced HSC contraction by interfering with the AT1R-mediated RhoA/ROCK2 pathway.

MeSH terms

  • Angiotensin II / pharmacology*
  • Animals
  • Carbon Tetrachloride Poisoning / enzymology
  • Carbon Tetrachloride Poisoning / pathology
  • Carvedilol / pharmacology*
  • Cell Proliferation / drug effects*
  • Hepatic Stellate Cells / enzymology*
  • Hepatic Stellate Cells / pathology
  • Humans
  • Liver Cirrhosis / enzymology
  • Liver Cirrhosis / pathology
  • Male
  • Mice
  • Receptor, Angiotensin, Type 1 / metabolism
  • rho-Associated Kinases / metabolism*
  • rhoA GTP-Binding Protein / metabolism*

Substances

  • Receptor, Angiotensin, Type 1
  • Carvedilol
  • Angiotensin II
  • RHOA protein, human
  • ROCK2 protein, human
  • Rock2 protein, mouse
  • rho-Associated Kinases
  • RhoA protein, mouse
  • rhoA GTP-Binding Protein