Role of TGF-beta1 in production of fibronectin in vascular smooth muscle cells cultured under high-phosphate conditions

Abstract

Hyperphosphatemia is associated with up-regulation of the extracellular matrix formation in vascular smooth muscle cells (VSMCs) and increased risk of cardiovascular disease. In the present study, the role of transforming growth factor-β1 (TGF-β1) in the production of fibronectin (FN) in vascular smooth muscle cells in high-phosphate environments was evaluated. Rat VSMCs were stimulated by high levels of phosphate or TGF-β1 in vitro. Levels of FN, TGF-β1, and TGF-β1 type I receptor (TβRI) proteins were measured by Western blot and immunostaining. Levels of active TGF-β1 in the supernatant of the culture medium were detected by ELISA. Production of FN was increased after VSMCs were incubated under high-phosphate conditions (2.5 mmol/L) for 12 hours. TGF-β1 (1 ng/mL) increased FN levels in cells as early as 3 hours after the start of treatment. Both TGF-β1 and TβRI were significantly up-regulated after 3-6 hours of stimulation with high phosphate. When VSMCs were pretreated with TGF-β1 neutralization antibody (10 µg/mL) for 30 minutes, induction of FN stimulated by high levels of phosphate was largely attenuated. TGF-β1 plays a critical role in the pathogenesis of high-phosphate-induced FN production in VSMCs in vitro.