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PMID: 19332549 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Regulation of Rac1 by simvastatin in endothelial cells: differential roles of AMP-activated protein kinase and calmodulin-dependent kinase kinase-beta.

The Journal of biological chemistry ·Vol. 284 ·No. 22 ·2009-05-29 ·Pages 14734-43

Kou R, Sartoretto J, Michel T

Abstract

These studies explore the connections between simvastatin, Rac1, and AMP-activated protein kinase (AMPK) pathways in cultured vascular endothelial cells and in arterial preparations isolated from statin-treated mice. In addition to their prominent effects on lipoprotein metabolism, statins can regulate the small GTPase Rac1, and may also affect the phosphorylation of the ubiquitous AMPK. We explored pathways of statin-modulated Rac1 and AMPK activation both in arterial preparations from statin-treated mice as well as in cultured endothelial cells. We treated adult mice with simvastatin daily for 2 weeks and then harvested and analyzed arterial preparations. Simvastatin treatment of mice led to a significant increase in AMPK and LKB1 phosphorylation and to a decrease in protein kinase A activity relative to control animals, associated with a marked increase in Rac1 activation. Exposure of bovine aortic endothelial cells to simvastatin for 24 h strikingly increased GTP-bound Rac1 and led to increased phosphorylation of AMPK as well as the AMPK kinase LKB1. These responses to simvastatin were blocked by mevalonate or geranylgeranyl pyrophosphate but not by farnesyl pyrophosphate. Small interfering RNA (siRNA)-mediated knockdown of AMPK abrogated simvastatin-induced Rac1 activation and LKB1 phosphorylation. Importantly, siRNA-mediated knockdown of the key AMPK kinase, calcium/calmodulin-dependent protein kinase kinase beta, completely blocked simvastatin-induced endothelial cell migration and also abrogated statin-promoted phosphorylation of AMPK and LKB1, as did pharmacological inhibition with the specific calcium/calmodulin-dependent protein kinase beta inhibitor STO-609. Moreover, siRNA-mediated knockdown of Rac1 completely blocked simvastatin-induced LKB1 phosphorylation, but without affecting simvastatin-induced AMPK phosphorylation. These findings establish a key role for simvastatin in activation of a novel Rac1-dependent signaling pathway in the vascular wall.

MeSH Terms
AMP-Activated Protein Kinases/metabolism Animals Arteries/drug effects,enzymology Calcium-Calmodulin-Dependent Protein Kinase Kinase/metabolism Cattle Cell Adhesion Molecules/metabolism Cell Movement/drug effects Cells, Cultured Endothelial Cells/cytology,drug effects,enzymology Enzyme Activation/drug effects Enzyme Inhibitors/pharmacology Gene Knockdown Techniques Humans Mice Microfilament Proteins/metabolism Molecular Mimicry/drug effects Phosphoproteins/metabolism Phosphorylation/drug effects Protein Serine-Threonine Kinases/metabolism RNA, Small Interfering/metabolism Simvastatin/pharmacology Terpenes rac1 GTP-Binding Protein/metabolism
Chemicals
Cell Adhesion Molecules Enzyme Inhibitors Microfilament Proteins Phosphoproteins RNA, Small Interfering Terpenes vasodilator-stimulated phosphoprotein Simvastatin Protein Serine-Threonine Kinases Stk11 protein, mouse Calcium-Calmodulin-Dependent Protein Kinase Kinase AMP-Activated Protein Kinases rac1 GTP-Binding Protein
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Kou Ruqin
Cardiovascular Division, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.
Sartoretto Juliano
Michel Thomas
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Article Info
Journal
The Journal of biological chemistry
Abbr.
J Biol Chem
ISSN
0021-9258
DOI
10.1074/jbc.M808664200
Published
2009-05-29
Epub
2009-00-30
Pages
14734-43
Language
English
Region
United States
NLM ID
2985121R
PMCID
PMC2685655
Subset
IM
Grants
NIGMS NIH HHS · GM36259 · United States
NHLBI NIH HHS · HL46457 · United States
NHLBI NIH HHS · HL48743 · United States
External Links
PubMed source DOI: 10.1074/jbc.M808664200 PMC Full Text View journal details
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