RhoA/Rho kinase (ROCK) alters fetuin-A uptake and regulates calcification in bovine vascular smooth muscle cells (BVSMC)

Neal X. Chen, Xianming Chen, Kalisha D. O'Neill, Simon J. Atkinson, Sharon M. Moe

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

RhoA/Rho kinases (ROCK) play a critical role in vascular smooth muscle cell (VSMC) actin cytoskeleton organization, differentiation, and function and are implicated in the pathogenesis of cardiovascular disease. We have previously determined that an important step in the regulation of calcification is fetuin-A endocytosis, a process that is dependent on changes in the cytoskeleton, which, in turn, is known to be affected by the RhoA/ ROCK signaling pathway. In the present study, bovine VSMC (BVSMC) were treated with the ROCK inhibitor Y-27632 or transfected with ROCK small interfering (si) RNA to knock down ROCK expression. Both conditions resulted in reduced actin stress fibers and increased Cy5-labeled fetuin-A uptake. Inhibition of ROCK by Y-27632 or siRNA also significantly increased BVSMC alkaline phosphatase (ALP) activity and calcification of BVSMC and rat aorta organ cultures. Cells were then incubated in calcification media in the presence or absence of Y-27632 and matrix vesicles (MV) isolated by collagenase digestion. These MV, isolated from BVSMC incubated with Y-27632, had increased ALP activity and increased ability of MV to subsequently calcify collagen by 66%. In contrast, activation of RhoA, which is upstream of ROCK, by transfecting plasmids encoding the dominant active Rho GTPase mutant (Rho-L63) led to decreased fetuin-A uptake and reduced calcification in BVSMC. These results demonstrate that the RhoA/ROCK signaling pathway is an important negative regulator of vascular calcification. vascular calcification; endocytosis; cytoskeleton; Rho GTPase.

Original languageEnglish (US)
Pages (from-to)F674-F680
JournalAmerican Journal of Physiology - Renal Physiology
Volume299
Issue number3
DOIs
StatePublished - Sep 1 2010

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alpha-2-HS-Glycoprotein
rho-Associated Kinases
Vascular Smooth Muscle
Smooth Muscle Myocytes
Vascular Calcification
rho GTP-Binding Proteins
Endocytosis
Cytoskeleton
Small Interfering RNA
Alkaline Phosphatase
Stress Fibers
Organ Culture Techniques
Collagenases
Actin Cytoskeleton
Aorta
Actins
Digestion
Plasmids
Cardiovascular Diseases
Collagen

ASJC Scopus subject areas

  • Physiology
  • Urology

Cite this

RhoA/Rho kinase (ROCK) alters fetuin-A uptake and regulates calcification in bovine vascular smooth muscle cells (BVSMC). / Chen, Neal X.; Chen, Xianming; O'Neill, Kalisha D.; Atkinson, Simon J.; Moe, Sharon M.

In: American Journal of Physiology - Renal Physiology, Vol. 299, No. 3, 01.09.2010, p. F674-F680.

Research output: Contribution to journalArticle

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