TNF-α down-regulates sarcoplasmic reticulum Ca2+ ATPase expression and leads to left ventricular diastolic dysfunction through binding of NF-κB to promoter response element

Chia Ti Tsai, Cho Kai Wu, Jen Kuang Lee, Sheng Nan Chang, Yu Min Kuo, Yi Chih Wang, Ling Ping Lai, Fu Tien Chiang, Juey Jen Hwang, Jiunn Lee Lin

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Aims TNF-alpha (TNF-α) causes left ventricular diastolic dysfunction. Down-regulation of sarcoplasmic reticulum Ca2+- ATPase 2a protein (SERCA2a) expression is one of the major mechanisms underlying diastolic dysfunction. We investigated whether TNF- α modulates SERCA2a expression and alters cardiac diastolic function, and its detailed signalling pathway. Methods and results We used both in vitro cellular cardiomyocyte model and in vivo rat model to address this issue. We found that TNF- α decreased the levels of both SERCA2a mRNA and protein in the cardiomyocytes, with corresponding impairment of diastolic calcium reuptake, a cellular phenotype of cardiac diastolic function. An -2 kb promoter of the SERCA2a gene (atp2a2) along with its serial deletions was cloned into the luciferase reporter system. TNF- α significantly decreased the promoter activity, and truncation of the SERCA2a gene promoter with the putative nuclear factor kappa-B (NF-κB) response element abolished TNF- α-induced SERCA2a gene suppression. Chromatin immunoprecipitation and gel retardation also confirmed the binding of NF-κB to this putative-binding site. TNF- α increased the phosphorylation of IKK and the degradation of IκB, resulted in NF-κB nuclear translocation, and decreasedSERCA2a gene promoter activity. This process was attenuated by NF-κB blockers and simvastatin. In the in vivo rat model, lipopolysaccharide treatment significantly elevated the serum TNF-α level, as well as phosphorylation of IKK, resulting in a decrease in myocardial SERCA2a expression, diastolic calcium reuptake, and diastolic dysfunction. Oral treatment with simvastatin led to an increase in SERCA2a expression, alleviation, and prevention of the diastolic dysfunction. Conclusions TNF-α suppresses SERCA2a gene expression via the IKK/IκB/NF-κB pathway and binding of NF-κB to the SERCA2a gene promoter, and its effect is blocked by simvastatin, demonstrating the potential therapeutic effect of statins in treating inflammation-related diastolic dysfunction.

Original languageEnglish
Pages (from-to)318-329
Number of pages12
JournalCardiovascular Research
Volume105
Issue number3
DOIs
Publication statusPublished - Jan 1 2015
Externally publishedYes

Fingerprint

Calcium-Transporting ATPases
NF-kappa B
Sarcoplasmic Reticulum
Response Elements
Left Ventricular Dysfunction
Down-Regulation
Simvastatin
Genes
Cardiac Myocytes
Phosphorylation
Calcium
Hydroxymethylglutaryl-CoA Reductase Inhibitors
Chromatin Immunoprecipitation
Therapeutic Uses
Luciferases
Lipopolysaccharides
Proteins
Tumor Necrosis Factor-alpha
Gels
Binding Sites

Keywords

  • Atp2a2
  • Diastolic dysfunction
  • SERCA2a
  • Simvastatin
  • TNF-α
  • Transcription

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine
  • Physiology (medical)

Cite this

TNF-α down-regulates sarcoplasmic reticulum Ca2+ ATPase expression and leads to left ventricular diastolic dysfunction through binding of NF-κB to promoter response element. / Tsai, Chia Ti; Wu, Cho Kai; Lee, Jen Kuang; Chang, Sheng Nan; Kuo, Yu Min; Wang, Yi Chih; Lai, Ling Ping; Chiang, Fu Tien; Hwang, Juey Jen; Lin, Jiunn Lee.

In: Cardiovascular Research, Vol. 105, No. 3, 01.01.2015, p. 318-329.

Research output: Contribution to journalArticle

Tsai, Chia Ti ; Wu, Cho Kai ; Lee, Jen Kuang ; Chang, Sheng Nan ; Kuo, Yu Min ; Wang, Yi Chih ; Lai, Ling Ping ; Chiang, Fu Tien ; Hwang, Juey Jen ; Lin, Jiunn Lee. / TNF-α down-regulates sarcoplasmic reticulum Ca2+ ATPase expression and leads to left ventricular diastolic dysfunction through binding of NF-κB to promoter response element. In: Cardiovascular Research. 2015 ; Vol. 105, No. 3. pp. 318-329.
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AU - Wu, Cho Kai

AU - Lee, Jen Kuang

AU - Chang, Sheng Nan

AU - Kuo, Yu Min

AU - Wang, Yi Chih

AU - Lai, Ling Ping

AU - Chiang, Fu Tien

AU - Hwang, Juey Jen

AU - Lin, Jiunn Lee

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N2 - Aims TNF-alpha (TNF-α) causes left ventricular diastolic dysfunction. Down-regulation of sarcoplasmic reticulum Ca2+- ATPase 2a protein (SERCA2a) expression is one of the major mechanisms underlying diastolic dysfunction. We investigated whether TNF- α modulates SERCA2a expression and alters cardiac diastolic function, and its detailed signalling pathway. Methods and results We used both in vitro cellular cardiomyocyte model and in vivo rat model to address this issue. We found that TNF- α decreased the levels of both SERCA2a mRNA and protein in the cardiomyocytes, with corresponding impairment of diastolic calcium reuptake, a cellular phenotype of cardiac diastolic function. An -2 kb promoter of the SERCA2a gene (atp2a2) along with its serial deletions was cloned into the luciferase reporter system. TNF- α significantly decreased the promoter activity, and truncation of the SERCA2a gene promoter with the putative nuclear factor kappa-B (NF-κB) response element abolished TNF- α-induced SERCA2a gene suppression. Chromatin immunoprecipitation and gel retardation also confirmed the binding of NF-κB to this putative-binding site. TNF- α increased the phosphorylation of IKK and the degradation of IκB, resulted in NF-κB nuclear translocation, and decreasedSERCA2a gene promoter activity. This process was attenuated by NF-κB blockers and simvastatin. In the in vivo rat model, lipopolysaccharide treatment significantly elevated the serum TNF-α level, as well as phosphorylation of IKK, resulting in a decrease in myocardial SERCA2a expression, diastolic calcium reuptake, and diastolic dysfunction. Oral treatment with simvastatin led to an increase in SERCA2a expression, alleviation, and prevention of the diastolic dysfunction. Conclusions TNF-α suppresses SERCA2a gene expression via the IKK/IκB/NF-κB pathway and binding of NF-κB to the SERCA2a gene promoter, and its effect is blocked by simvastatin, demonstrating the potential therapeutic effect of statins in treating inflammation-related diastolic dysfunction.

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KW - Transcription

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KW - Diastolic dysfunction

KW - SERCA2a

KW - Simvastatin

KW - TNF-α

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