Angiotensin II Suppresses Rev-erbα Expression in THP-1 Macrophages via the Ang II Type 1 Receptor/Liver X Receptor α Pathway

Cell Physiol Biochem. 2018;46(1):303-313. doi: 10.1159/000488431. Epub 2018 Mar 22.

Abstract

Background/aims: Angiotensin II (Ang II) regulates the expression of some core clock genes; excess Ang II leads to atherosclerosis advancement. Macrophage Rev-erbα mediates clockwork and inflammation, and plays a role in atherosclerotic lesion progression. However, the role of Ang II in regulating Rev-erbα expression in macrophages remains unclarified.

Methods: We induced THP-1 macrophages by phorbol 12-myristate 13-acetate and investigated the effect of Ang II on Rev-erbα expression via real-time polymerase chain reaction, western blotting and small interfering RNA (siRNA) techniques. The cytotoxicity of the Rev-erbα agonist SR9009 was analyzed using a (3-[4,5-dimethylthiazol-2-yl])-2,5- diphenyltetrazolium bromide assay.

Results: Ang II suppressed Rev-erbα mRNA and protein expression in THP-1 macrophages in a dose and time dependent manner. This effect was mediated via Ang II type 1 receptor (AT1R), and not Ang II type 2 receptor or peroxisome proliferator-activated receptor γ (PPARγ). Consistent with Rev-erbα expression regulated by Ang II, the liver X receptor α (LXRα) protein expression was downregulated in a time-dependent manner after Ang II treatment. The activation or silence of LXRα significantly increased or decreased Rev-erbα expression regulated by Ang II, respectively. This suggests that LXRα is involved in the effect of Ang II on Rev-erbα expression. MMP-9 mRNA expressions were significantly suppressed by SR9009 in THP-1 and RAW264.7 macrophages; moreover, SR9009-treatment significantly reduced Ang II-induced MMP-9 protein expressions in two types of macrophages.

Conclusion: Ang II downregulates Rev-erbα expression in THP-1 macrophages via the AT1R/LXRα pathway.

Keywords: Angiotensin II; Liver X receptor; Macrophages; Matrix metalloproteinase-9; Rev-erbα.

MeSH terms

  • Angiotensin II / pharmacology*
  • Animals
  • Cell Line
  • Down-Regulation / drug effects*
  • Humans
  • Imidazoles / pharmacology
  • Liver X Receptors / antagonists & inhibitors
  • Liver X Receptors / genetics
  • Liver X Receptors / metabolism*
  • Macrophages / cytology
  • Macrophages / drug effects
  • Macrophages / metabolism
  • Matrix Metalloproteinase 9 / metabolism
  • Mice
  • Nuclear Receptor Subfamily 1, Group D, Member 1 / agonists
  • Nuclear Receptor Subfamily 1, Group D, Member 1 / genetics
  • Nuclear Receptor Subfamily 1, Group D, Member 1 / metabolism*
  • Pyridines / pharmacology
  • RAW 264.7 Cells
  • RNA Interference
  • RNA, Small Interfering / metabolism
  • Receptor, Angiotensin, Type 1 / chemistry
  • Receptor, Angiotensin, Type 1 / metabolism*
  • Signal Transduction / drug effects*
  • Valsartan / pharmacology

Substances

  • Imidazoles
  • Liver X Receptors
  • NR1D1 protein, human
  • Nuclear Receptor Subfamily 1, Group D, Member 1
  • Pyridines
  • RNA, Small Interfering
  • Receptor, Angiotensin, Type 1
  • Angiotensin II
  • PD 123319
  • Valsartan
  • Matrix Metalloproteinase 9