Cyclophilin A is secreted by a vesicular pathway in vascular smooth muscle cells

Jun Suzuki, Zheng Gen Jin, David F. Meoli, Tetsuya Matoba, Bradford C. Berk

Research output: Contribution to journalArticle

144 Citations (Scopus)

Abstract

Reactive oxygen species (ROS) contribute to the pathogenesis of atherosclerosis in part by promoting vascular smooth muscle cell (VSMC) growth. Previously we demonstrated that cyclophilin A (CyPA) is a secreted oxidative stress-induced factor (SOXF) that promotes inflammation, VSMC growth, and endothelial cell apoptosis. However, the mechanisms that regulate CyPA secretion are unknown. In this study, we hypothesized that ROS-induced CyPA secretion from VSMC requires a highly regulated process of vesicle transport, docking, and fusion at the plasma membrane. Conditioned medium and plasma membrane sheets were prepared by exposing VSMC to 1 μmol/L LY83583, which generates intracellular superoxide. A vesicular transport mechanism was confirmed by colocalization at the plasma membrane with vesicle-associated membrane protein (VAMP). CyPA transport to the plasma membrane and secretion were significantly increased by LY83583. Reduction of VAMP-2 expression by small interfering RNA inhibited LY83583-induced CyPA secretion. Pretreatment with 3 μmol/L cytochalasin D, an actin depolymerizing agent, abrogated CyPA secretion. Infection with dominant-negative RhoA and Cdc42 adenovirus inhibited CyPA secretion by 72% and 63%, respectively, whereas dominant-negative Rac1 had a small effect (11%). Pretreatment with the Rho kinase inhibitor Y27632 (3 to 30 μmol/L) and myosin II inhibitor blebbistatin (1 to 10 μmol/L) inhibited CyPA secretion in a dose-dependent manner. Simvastatin (3 to 30 μmol/L) also dose-dependently inhibited LY83583-induced CyPA secretion likely via decreased isoprenylation of small GTPases. Our findings define a novel VSMC vesicular secretory pathway for CyPA that involves actin remodeling and myosin II activation via RhoA-, Cdc42-, and Rho kinase-dependent signaling events.

Original languageEnglish
Pages (from-to)811-817
Number of pages7
JournalCirculation research
Volume98
Issue number6
DOIs
Publication statusPublished - Mar 1 2006
Externally publishedYes

Fingerprint

Cyclophilin A
Vascular Smooth Muscle
Smooth Muscle Myocytes
6-anilino-5,8-quinolinedione
Cell Membrane
Myosin Type II
rho-Associated Kinases
Actins
Reactive Oxygen Species
Vesicle-Associated Membrane Protein 2
R-SNARE Proteins
Prenylation
Cytochalasin D
Transport Vesicles
Simvastatin
Monomeric GTP-Binding Proteins
Secretory Pathway
Conditioned Culture Medium
Growth
Adenoviridae

All Science Journal Classification (ASJC) codes

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Cyclophilin A is secreted by a vesicular pathway in vascular smooth muscle cells. / Suzuki, Jun; Jin, Zheng Gen; Meoli, David F.; Matoba, Tetsuya; Berk, Bradford C.

In: Circulation research, Vol. 98, No. 6, 01.03.2006, p. 811-817.

Research output: Contribution to journalArticle

Suzuki, Jun ; Jin, Zheng Gen ; Meoli, David F. ; Matoba, Tetsuya ; Berk, Bradford C. / Cyclophilin A is secreted by a vesicular pathway in vascular smooth muscle cells. In: Circulation research. 2006 ; Vol. 98, No. 6. pp. 811-817.
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abstract = "Reactive oxygen species (ROS) contribute to the pathogenesis of atherosclerosis in part by promoting vascular smooth muscle cell (VSMC) growth. Previously we demonstrated that cyclophilin A (CyPA) is a secreted oxidative stress-induced factor (SOXF) that promotes inflammation, VSMC growth, and endothelial cell apoptosis. However, the mechanisms that regulate CyPA secretion are unknown. In this study, we hypothesized that ROS-induced CyPA secretion from VSMC requires a highly regulated process of vesicle transport, docking, and fusion at the plasma membrane. Conditioned medium and plasma membrane sheets were prepared by exposing VSMC to 1 μmol/L LY83583, which generates intracellular superoxide. A vesicular transport mechanism was confirmed by colocalization at the plasma membrane with vesicle-associated membrane protein (VAMP). CyPA transport to the plasma membrane and secretion were significantly increased by LY83583. Reduction of VAMP-2 expression by small interfering RNA inhibited LY83583-induced CyPA secretion. Pretreatment with 3 μmol/L cytochalasin D, an actin depolymerizing agent, abrogated CyPA secretion. Infection with dominant-negative RhoA and Cdc42 adenovirus inhibited CyPA secretion by 72{\%} and 63{\%}, respectively, whereas dominant-negative Rac1 had a small effect (11{\%}). Pretreatment with the Rho kinase inhibitor Y27632 (3 to 30 μmol/L) and myosin II inhibitor blebbistatin (1 to 10 μmol/L) inhibited CyPA secretion in a dose-dependent manner. Simvastatin (3 to 30 μmol/L) also dose-dependently inhibited LY83583-induced CyPA secretion likely via decreased isoprenylation of small GTPases. Our findings define a novel VSMC vesicular secretory pathway for CyPA that involves actin remodeling and myosin II activation via RhoA-, Cdc42-, and Rho kinase-dependent signaling events.",
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AU - Berk, Bradford C.

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