Monocrotaline pyrrole induces Smad nuclear accumulation and altered signaling expression in human pulmonary arterial endothelial cells

M. Ramos, M. W. Lamé, H. J. Segall, Dennis W Wilson

Research output: Contribution to journalArticle

18 Scopus citations

Abstract

The mechanistic relationship between the widely used monocrotaline model of primary pulmonary hypertension and altered TGFβ family signaling due to genetic defects in the Bone Morphogenetic Protein type II receptor in affected humans has not been investigated. In this study we use fluorescent microscopy to demonstrate nuclear translocation of Smad 4 in human pulmonary arterial endothelial cell (HPAEC) cultures treated with monocrotaline pyrrole (MCTP), Bone Morphogenetic Protein (BMP) and TGFβ. While MCTP induced transient nuclear accumulation of phosphorylated Smad 1 (P-Smad 1) and phosphorylated Smad 2 (P-Smad 2), only expression of P-Smad 1 was significantly altered in western blots. P-Smad 1 expression significantly increased 30 min following treatment with MCTP correlating with P-Smad 1 and Smad 4 nuclear translocation. Although a modest, but significant decrease in P-Smad 1 expression occurred 1 h after treatment, expression was significantly increased at 72 h. Evaluation of components of the signal and response pathway at 72 h showed decreased expression of the BMP type II receptor (BMPrII), no change in TGFβ Activin Receptor-like Kinase 1 (Alk 1), no change in Smad 4 but increase in the inhibitory Smad 6, decrease in the alternate BMP signaling pathway p38MAPK but no change in the psmad1 response element ID 1. Our results suggest transient activation of Smad signaling pathways in initial MCTP endothelial cell toxicity, and a persistent dysregulation of BMP signaling. Electron microscopy of cell membrane caveoli revealed a dramatic decrease in these structures after 72 h. Loss of these structural elements, noted for their sequestration and inhibition of receptor activity, may contribute to prolonged alterations in BMP signaling.

Original languageEnglish (US)
Pages (from-to)439-448
Number of pages10
JournalVascular Pharmacology
Volume46
Issue number6
DOIs
StatePublished - Jun 2007

Keywords

  • BMPrII
  • Bone morphogenic protein
  • MCTP
  • Monocrotaline pyrrole
  • Smad signal transduction

ASJC Scopus subject areas

  • Pharmacology

Fingerprint Dive into the research topics of 'Monocrotaline pyrrole induces Smad nuclear accumulation and altered signaling expression in human pulmonary arterial endothelial cells'. Together they form a unique fingerprint.

  • Cite this