Attenuation of endothelin-1-induced calcium response by tyrosine kinase inhibitors in vascular smooth muscle cells

Cynthia Y. Liu, Michael Sturek

Research output: Contribution to journalArticlepeer-review

30 Scopus citations

Abstract

Although tyrosine kinases play an important role in cell growth and have been implicated in regulation of smooth muscle contraction, their role in agonist-induced myoplasmic Ca2+ responses is unclear. We examined effects of the tyrosine kinase inhibitors genistein and methyl 2,5-dihydroxycinnamate (MDHC) on the endothelin-1 (ET-1)-induced Ca2+ response and determined underlying mechanisms for the effects. Freshly isolated smooth muscle cells from porcine coronary arteries were loaded with fura 2 ester, and myoplasmic free Ca2+ (Ca(m)/2+) concentration was estimated with fura 2 microfluorometry. Both genistein and MDHC inhibited the initial transient Ca(m)/2+ response to ET by 54 and 81%, respectively (P < 0.05), in the presence of extracellular Ca2+. Genistein also significantly delayed the Ca(m)/2+ response, with the latent period from ET-1 application to the beginning of the Ca(m)/2+ response being increased from 1.08 ± 0.17 to 2.65 ± 0.52 min (P < 0.05). In the absence of extracellular Ca2+, genistein inhibited the ET-1-induced Ca(m)/2+ response by 93% (P < 0.05). The Ca(m)/2+ responses to caffeine (5 mM) or inositol trisphosphate (IP3) applied intracellularly via a patch-clamp pipette were not affected by genistein. Both genistein and MDHC also abolished the sustained Ca(m)/2+ response to ET-1. However, the Ca(m)/2+ response to depolarization by 80 mM K+ was not inhibited by MDHC and only inhibited 22% by genistein (P < 0.05). These results indicate that 1) activation of tyrosine kinases is an important regulatory mechanism for the ET-1-induced Ca(m)/2+ response in vascular smooth muscle and 2) tyrosine kinases mediate ET-1-induced Ca2+ release with no direct effect on IP3-mediated Ca2+ release. Thus ET-1-mediated signaling upstream of IP3 interaction with the Ca2+ stores is regulated by tyrosine kinases.

Original languageEnglish (US)
Pages (from-to)C1825-C1833
JournalAmerican Journal of Physiology - Cell Physiology
Volume270
Issue number6 39-6
DOIs
StatePublished - Jun 1996

Keywords

  • caffeine
  • fura 2
  • genistein
  • inositol trisphosphate
  • methyl 2,5-dihydroxycinnamate
  • sarcoplasmic reticulum
  • whole cell current

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Cell Biology
  • Physiology
  • Physiology (medical)

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