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Inhibition of JAK/STAT signaling pathway prevents high-glucose-induced increase in endothelin-1 synthesis in human endothelial cells

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Abstract

Emerging evidence demonstrates the involvement of endothelin-1 (ET-1) in the pathophysiology of cardiovascular disorders associated with diabetes mellitus. The molecular mechanisms accountable for the increased production of ET-1 are not completely defined. The Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway is an essential pathogenic mechanism leading to endothelial cell dysfunction. Our aim has been to investigate the role of JAK/STAT in the regulation of ET-1 synthesis in human endothelial cells (EAhy926 cells line). EAhy926 cells were exposed to normal (5 mM) or high (25 mM) glucose concentrations in the presence/absence of various JAK/STAT inhibitors. Using real-time polymerase chain reaction, enzyme-linked immunosorbent assay, and gene reporter assay, we found that JAK/STAT inhibitors (STAT1 decoy oligodeoxynucleotides, AG490, S3I201, WP1066) significantly diminished the high-glucose-dependent up-regulation of ET-1 mRNA, peptide synthesis, and promoter activity. In silico analysis of the human ET-1 promoter revealed the presence of typical STAT1-gamma-activated sequence (STAT1-GAS) elements. Transient overexpression of STAT1 indicated an up-regulation of ET-1 promoter activity. Chromatin immunoprecipitation demonstrated the physical interaction of STAT1 proteins with the predicted GAS sites. Regulation of ET-1 synthesis by the JAK/STAT pathway thus represents a novel mechanism by which high glucose induces endothelial cell dysfunction in diabetes. Since the JAK/STAT system is an important regulator of the response of endothelial cells to injury, the modulation of this system and the subsequent decrease in ET-1 level may represent a key pharmacological target in diabetes-associated cardiovascular disorders.

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Acknowledgments

The authors thank Prof. Dr. Angelika Bierhaus from the University of Heidelberg, Germany for the plasmid containing the human endothelin-1 gene promoter and acknowledge the skilful assistance of Floarea Georgescu, Ioana Manolescu, and Constanta Stan.

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Authors and Affiliations

  1. Institute of Cellular Biology and Pathology ‘Nicolae Simionescu’, 8 BP Hasdeu Street, 050568, Bucharest, Romania

    Simona-Adriana Manea, Adrian Manea & Constantina Heltianu

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  1. Simona-Adriana Manea
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  2. Adrian Manea
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  3. Constantina Heltianu
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Correspondence to Constantina Heltianu.

Additional information

This work was supported by grants from the Romanian Academy (65/2007) and Romanian Ministry of Education, Research, and Innovation (100/2007).

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Manea, SA., Manea, A. & Heltianu, C. Inhibition of JAK/STAT signaling pathway prevents high-glucose-induced increase in endothelin-1 synthesis in human endothelial cells. Cell Tissue Res 340, 71–79 (2010). https://doi.org/10.1007/s00441-010-0936-1

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  • DOI: https://doi.org/10.1007/s00441-010-0936-1

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