LPS-induced TNF-α release from and apoptosis in rat cardiomyocytes: Obligatory role for CD14 in mediating the LPS response

Kevyn L. Comstock, Kevin A. Krown, M. Trevor Page, Don Martin, Peter Ho, Melinda Pedraza, Ernesto N. Castro, Nobuko Nakajima, Christopher C. Glembotski, P. J.E. Quintana, Roger A. Sabbadini

Research output: Contribution to journalArticlepeer-review

135 Scopus citations

Abstract

The bacterial endotoxin lipopolysaccharide (LPS) contributes to the cardiovascular collapse and death observed in patients with sepsis. Because LPS has such profound effects on cardiac performance, we speculate that direct effects of LPS could be demonstrated on cardiomyocytes in culture, and that these direct effects are mediated by the LPS receptor, CD14. Accordingly, in this study, we provide evidence for CD14-dependent cardiotoxic effects of LPS including the LPS-stimulated secretion of tumor necrosis factor alpha (TNF-α) from cardiomyocytes. TNF-α is an inflammatory cytokine which is known for its negative inotropic effects on cardiac performance, but has not until recently been shown to be produced by cardiac cells. In this study, LPS was found to stimulate strongly in a dose-dependent manner the secretion of TNF-α from cultured adult rat cardiomyocytes. Further, LPS-induced TNF-α secretion was blocked by an inhibitor of TNF-α processing, metallomatrix protease inhibitor (TAPI). Molecular and immunological evidence demonstrated the presence of LPS receptors (CD14) on cardiomyocytes. Attenuated TNF-α secretion following PI-PLC treatment confirmed the functional importance of CD14 for LPS-mediated myocardial effects. Importantly, LPS also triggered apoptosis in cultured cardiomyocytes as quantified by single-cell gel electrophoresis of nuclei exhibiting DNA fragmentation patterns characteristic of apoptosis (i.e. cardiac comets). Apoptotic cell death was blocked by pre-incubation with the soluble TNF-α receptor fragment (TNFRII:Fc), suggesting that LPS-induced apoptosis was TNF-α-dependent and probably involved an autocrine function for the TNF-α whose secretion was under LPS control. The results of this study suggest that the cardiodepressant effects of LPS are dependent on CD14 signaling and may not only be due to acute negative inotropic effects of TNF-α but also may be complicated by TNF-α-induced apoptotic cell death which effectively reduces the number of working myocardial cells.

Original languageEnglish (US)
Pages (from-to)2761-2775
Number of pages15
JournalJournal of Molecular and Cellular Cardiology
Volume30
Issue number12
DOIs
StatePublished - Dec 1998
Externally publishedYes

Keywords

  • Apoptosis
  • CD14
  • Lipopolysaccharide
  • Sepsis
  • TNF-α

ASJC Scopus subject areas

  • Molecular Biology
  • Cardiology and Cardiovascular Medicine

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