Redox Regulation of Vascular Smooth Muscle Cell Differentiation

• Published in: Circulation research Vol. 89; no. 1; pp. 39 - 46
• Main Authors: Su, B, Mitra, S, Gregg, H, Flavahan, S, Chotani, M A, Clark, K R, Goldschmidt-Clermont, P J, Flavahan, N A
• Format: Journal Article
• Language: English
• Published: Hagerstown, MD American Heart Association, Inc 06-07-2001; Lippincott; Lippincott Williams & Wilkins Ovid Technologies
• Subjects: Acetylcysteine - pharmacology; Biological and medical sciences; Blood vessels and receptors; Calcium-Binding Proteins - metabolism; Calponins; Catalase - pharmacology; Cell Differentiation; Cell Division; Cells, Cultured; Fundamental and applied biological sciences. Psychology; Humans; MAP Kinase Signaling System; Metalloporphyrins - pharmacology; Microfilament Proteins; Mitogen-Activated Protein Kinases - physiology; Muscle, Smooth, Vascular - cytology; Muscle, Smooth, Vascular - drug effects; Muscle, Smooth, Vascular - physiology; Myosins - metabolism; Oxidation-Reduction; p38 Mitogen-Activated Protein Kinases; Phenotype; Reactive Oxygen Species - physiology; Vertebrates: cardiovascular system; Human; Redox state; Oxygen; Enzyme; Transferases; Smooth muscle; In vitro; Artery; Free radical; Signal transduction; Regulation(control); Protein kinase; Blood vessel; Myosin; Aorta; Circulatory system; Differentiation
• ISSN: 0009-7330; 1524-4571
• DOI: 10.1161/hh1301.093615

ABSTRACT—Experiments were performed to determine the role of reactive oxygen species (ROS) in regulating vascular smooth muscle cell (VSMC) phenotype. After quiescence, cultured human VSMCs increased their expression of differentiation proteins (α-actin, calponin, and SM1 and SM2 myosin), but not β-actin. ROS activity, determined using the H2O2-sensitive probe dichlorodihydrofluorescein (DCF), remained high in quiescent cells and was inhibited by catalase (3000 U/mL) or by N-acetylcysteine (NAC, 2 to 20 mmol/L). A superoxide dismutase mimic (SOD; MnTMPyP, 25 μmol/L) or SOD plus low concentrations of NAC (SODNAC2, 2 mmol/L) increased DCF fluorescence, which was inhibited by catalase or by NAC (10 to 20 mmol/L). Inhibition of ROS activity (by catalase or NAC) decreased the baseline expression of differentiation proteins, whereas elevation of ROS (by SOD or SODNAC2) increased expression of the differentiation markers. The latter effect was blocked by catalase or by NAC (10 to 20 mmol/L). None of the treatments altered β-actin expression. SODNAC2-treated cells demonstrated contractions to endothelin that were absent in proliferating cells. p38 Mitogen-activated protein kinase (MAPK) activity was decreased when ROS activity was reduced (NAC, 10 mmol/L) and was augmented when ROS activity was increased (SODNAC2). Inhibition of p38 MAPK with pyridyl imidazole compound (SB202190, 2 to 10 μmol/L) reduced expression of differentiation proteins occurring under basal conditions and in response to SODNAC2. Transduction of VSMCs with an adenovirus encoding constitutively active MKK6, an activator of p38 MAPK, increased expression of differentiation proteins, whereas transduction with an adenovirus encoding dominant-negative p38 MAPK decreased expression of the differentiation proteins. These findings demonstrate that ROS can increase VSMC differentiation through a p38 MAPK–dependent pathway.