Homocysteine affects cardiomyocyte viability: concentration-dependent effects on reversible flip-flop, apoptosis and necrosis

Jessica A Sipkens; Paul A J Krijnen; Christof Meischl; Saskia A G M Cillessen; Yvo M Smulders; Desirée E C Smith; Cindy P E Giroth; Marieke D Spreeuwenberg; René J P Musters; Alice Muller; Cornelis Jakobs; Dirk Roos; Coen D A Stehouwer; Jan A Rauwerda; Victor W M van Hinsbergh; Hans W M Niessen

doi:https://doi.org/10.1007/s10495-007-0077-5

Homocysteine affects cardiomyocyte viability: concentration-dependent effects on reversible flip-flop, apoptosis and necrosis

Jessica A Sipkens, Paul A J Krijnen, Christof Meischl, Saskia A G M Cillessen, Yvo M Smulders, Desirée E C Smith, Cindy P E Giroth, Marieke D Spreeuwenberg, René J P Musters, Alice Muller, Cornelis Jakobs, Dirk Roos, Coen D A Stehouwer, Jan A Rauwerda, Victor W M van Hinsbergh, Hans W M Niessen

Research output: Contribution to journal › Article › Academic › peer-review

42 Citations (Scopus)

Abstract

BACKGROUND: Hyperhomocysteinaemia (HHC) is thought to be a risk factor for cardiovascular disease including heart failure. While numerous studies have analyzed the role of homocysteine (Hcy) in the vasculature, only a few studies investigated the role of Hcy in the heart. Therefore we have analyzed the effects of Hcy on isolated cardiomyocytes.

METHODS: H9c2 cells (rat cardiomyoblast cells) and adult rat cardiomyocytes were incubated with Hcy and were analyzed for cell viability. Furthermore, we determined the effects of Hcy on intracellular mediators related to cell viability in cardiomyocytes, namely NOX2, reactive oxygen species (ROS), mitochondrial membrane potential (DeltaPsi (m)) and ATP concentrations.

RESULTS: We found that incubation of H9c2 cells with 0.1 mM D,L-Hcy (= 60 microM L-Hcy) resulted in an increase of DeltaPsi (m) as well as ATP concentrations. 1.1 mM D,L-Hcy (= 460 microM L-Hcy) induced reversible flip-flop of the plasma membrane phospholipids, but not apoptosis. Incubation with 2.73 mM D,L-Hcy (= 1.18 mM L-Hcy) induced apoptosis and necrosis. This loss of cell viability was accompanied by a thread-to-grain transition of the mitochondrial reticulum, ATP depletion and nuclear NOX2 expression coinciding with ROS production as evident from the presence of nitrotyrosin residues. Notably, only at this concentration we found a significant increase in S-adenosylhomocysteine which is considered the primary culprit in HHC.

CONCLUSION: We found concentration-dependent effects of Hcy in cardiomyocytes, varying from induction of reversible flip-flop of the plasma membrane phospholipids, to apoptosis and necrosis.

Original language	English
Pages (from-to)	1407-18
Number of pages	12
Journal	Apoptosis
Volume	12
Issue number	8
DOIs	https://doi.org/10.1007/s10495-007-0077-5
Publication status	Published - Aug 2007

Keywords

Adenosine Triphosphate
Animals
Apoptosis
Caspase 3
Cell Membrane
Cell Survival
Cells, Cultured
Dose-Response Relationship, Drug
Gene Expression Regulation
Homocysteine
Journal Article
Membrane Fluidity
Membrane Glycoproteins
Membrane Potential, Mitochondrial
Mitochondria, Heart
Models, Biological
Myocytes, Cardiac
NADPH Oxidase 2
NADPH Oxidases
Necrosis
Phospholipids
Protein Processing, Post-Translational
Rats
Research Support, Non-U.S. Gov't
S-Adenosylhomocysteine
S-Adenosylmethionine

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https://doi.org/10.1007/s10495-007-0077-5

Cite this

Sipkens, J. A., Krijnen, P. A. J., Meischl, C., Cillessen, S. A. G. M., Smulders, Y. M., Smith, D. E. C., Giroth, C. P. E., Spreeuwenberg, M. D., Musters, R. J. P., Muller, A., Jakobs, C., Roos, D., Stehouwer, C. D. A., Rauwerda, J. A., van Hinsbergh, V. W. M., & Niessen, H. W. M. (2007). Homocysteine affects cardiomyocyte viability: concentration-dependent effects on reversible flip-flop, apoptosis and necrosis. Apoptosis, 12(8), 1407-18. https://doi.org/10.1007/s10495-007-0077-5

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title = "Homocysteine affects cardiomyocyte viability: concentration-dependent effects on reversible flip-flop, apoptosis and necrosis",

abstract = "BACKGROUND: Hyperhomocysteinaemia (HHC) is thought to be a risk factor for cardiovascular disease including heart failure. While numerous studies have analyzed the role of homocysteine (Hcy) in the vasculature, only a few studies investigated the role of Hcy in the heart. Therefore we have analyzed the effects of Hcy on isolated cardiomyocytes.METHODS: H9c2 cells (rat cardiomyoblast cells) and adult rat cardiomyocytes were incubated with Hcy and were analyzed for cell viability. Furthermore, we determined the effects of Hcy on intracellular mediators related to cell viability in cardiomyocytes, namely NOX2, reactive oxygen species (ROS), mitochondrial membrane potential (DeltaPsi (m)) and ATP concentrations.RESULTS: We found that incubation of H9c2 cells with 0.1 mM D,L-Hcy (= 60 microM L-Hcy) resulted in an increase of DeltaPsi (m) as well as ATP concentrations. 1.1 mM D,L-Hcy (= 460 microM L-Hcy) induced reversible flip-flop of the plasma membrane phospholipids, but not apoptosis. Incubation with 2.73 mM D,L-Hcy (= 1.18 mM L-Hcy) induced apoptosis and necrosis. This loss of cell viability was accompanied by a thread-to-grain transition of the mitochondrial reticulum, ATP depletion and nuclear NOX2 expression coinciding with ROS production as evident from the presence of nitrotyrosin residues. Notably, only at this concentration we found a significant increase in S-adenosylhomocysteine which is considered the primary culprit in HHC.CONCLUSION: We found concentration-dependent effects of Hcy in cardiomyocytes, varying from induction of reversible flip-flop of the plasma membrane phospholipids, to apoptosis and necrosis.",

keywords = "Adenosine Triphosphate, Animals, Apoptosis, Caspase 3, Cell Membrane, Cell Survival, Cells, Cultured, Dose-Response Relationship, Drug, Gene Expression Regulation, Homocysteine, Journal Article, Membrane Fluidity, Membrane Glycoproteins, Membrane Potential, Mitochondrial, Mitochondria, Heart, Models, Biological, Myocytes, Cardiac, NADPH Oxidase 2, NADPH Oxidases, Necrosis, Phospholipids, Protein Processing, Post-Translational, Rats, Research Support, Non-U.S. Gov't, S-Adenosylhomocysteine, S-Adenosylmethionine",

author = "Sipkens, {Jessica A} and Krijnen, {Paul A J} and Christof Meischl and Cillessen, {Saskia A G M} and Smulders, {Yvo M} and Smith, {Desir{\'e}e E C} and Giroth, {Cindy P E} and Spreeuwenberg, {Marieke D} and Musters, {Ren{\'e} J P} and Alice Muller and Cornelis Jakobs and Dirk Roos and Stehouwer, {Coen D A} and Rauwerda, {Jan A} and {van Hinsbergh}, {Victor W M} and Niessen, {Hans W M}",

year = "2007",

month = aug,

doi = "https://doi.org/10.1007/s10495-007-0077-5",

language = "English",

volume = "12",

pages = "1407--18",

journal = "Apoptosis",

issn = "1360-8185",

publisher = "Springer Netherlands",

number = "8",

}

Sipkens, JA, Krijnen, PAJ, Meischl, C, Cillessen, SAGM , Smulders, YM , Smith, DEC, Giroth, CPE, Spreeuwenberg, MD, Musters, RJP, Muller, A , Jakobs, C, Roos, D, Stehouwer, CDA, Rauwerda, JA, van Hinsbergh, VWM & Niessen, HWM 2007, 'Homocysteine affects cardiomyocyte viability: concentration-dependent effects on reversible flip-flop, apoptosis and necrosis', Apoptosis, vol. 12, no. 8, pp. 1407-18. https://doi.org/10.1007/s10495-007-0077-5

TY - JOUR

T1 - Homocysteine affects cardiomyocyte viability

T2 - concentration-dependent effects on reversible flip-flop, apoptosis and necrosis

AU - Sipkens, Jessica A

AU - Krijnen, Paul A J

AU - Meischl, Christof

AU - Cillessen, Saskia A G M

AU - Smulders, Yvo M

AU - Smith, Desirée E C

AU - Giroth, Cindy P E

AU - Spreeuwenberg, Marieke D

AU - Musters, René J P

AU - Muller, Alice

AU - Jakobs, Cornelis

AU - Roos, Dirk

AU - Stehouwer, Coen D A

AU - Rauwerda, Jan A

AU - van Hinsbergh, Victor W M

AU - Niessen, Hans W M

PY - 2007/8

Y1 - 2007/8

N2 - BACKGROUND: Hyperhomocysteinaemia (HHC) is thought to be a risk factor for cardiovascular disease including heart failure. While numerous studies have analyzed the role of homocysteine (Hcy) in the vasculature, only a few studies investigated the role of Hcy in the heart. Therefore we have analyzed the effects of Hcy on isolated cardiomyocytes.METHODS: H9c2 cells (rat cardiomyoblast cells) and adult rat cardiomyocytes were incubated with Hcy and were analyzed for cell viability. Furthermore, we determined the effects of Hcy on intracellular mediators related to cell viability in cardiomyocytes, namely NOX2, reactive oxygen species (ROS), mitochondrial membrane potential (DeltaPsi (m)) and ATP concentrations.RESULTS: We found that incubation of H9c2 cells with 0.1 mM D,L-Hcy (= 60 microM L-Hcy) resulted in an increase of DeltaPsi (m) as well as ATP concentrations. 1.1 mM D,L-Hcy (= 460 microM L-Hcy) induced reversible flip-flop of the plasma membrane phospholipids, but not apoptosis. Incubation with 2.73 mM D,L-Hcy (= 1.18 mM L-Hcy) induced apoptosis and necrosis. This loss of cell viability was accompanied by a thread-to-grain transition of the mitochondrial reticulum, ATP depletion and nuclear NOX2 expression coinciding with ROS production as evident from the presence of nitrotyrosin residues. Notably, only at this concentration we found a significant increase in S-adenosylhomocysteine which is considered the primary culprit in HHC.CONCLUSION: We found concentration-dependent effects of Hcy in cardiomyocytes, varying from induction of reversible flip-flop of the plasma membrane phospholipids, to apoptosis and necrosis.

AB - BACKGROUND: Hyperhomocysteinaemia (HHC) is thought to be a risk factor for cardiovascular disease including heart failure. While numerous studies have analyzed the role of homocysteine (Hcy) in the vasculature, only a few studies investigated the role of Hcy in the heart. Therefore we have analyzed the effects of Hcy on isolated cardiomyocytes.METHODS: H9c2 cells (rat cardiomyoblast cells) and adult rat cardiomyocytes were incubated with Hcy and were analyzed for cell viability. Furthermore, we determined the effects of Hcy on intracellular mediators related to cell viability in cardiomyocytes, namely NOX2, reactive oxygen species (ROS), mitochondrial membrane potential (DeltaPsi (m)) and ATP concentrations.RESULTS: We found that incubation of H9c2 cells with 0.1 mM D,L-Hcy (= 60 microM L-Hcy) resulted in an increase of DeltaPsi (m) as well as ATP concentrations. 1.1 mM D,L-Hcy (= 460 microM L-Hcy) induced reversible flip-flop of the plasma membrane phospholipids, but not apoptosis. Incubation with 2.73 mM D,L-Hcy (= 1.18 mM L-Hcy) induced apoptosis and necrosis. This loss of cell viability was accompanied by a thread-to-grain transition of the mitochondrial reticulum, ATP depletion and nuclear NOX2 expression coinciding with ROS production as evident from the presence of nitrotyrosin residues. Notably, only at this concentration we found a significant increase in S-adenosylhomocysteine which is considered the primary culprit in HHC.CONCLUSION: We found concentration-dependent effects of Hcy in cardiomyocytes, varying from induction of reversible flip-flop of the plasma membrane phospholipids, to apoptosis and necrosis.

KW - Adenosine Triphosphate

KW - Animals

KW - Apoptosis

KW - Caspase 3

KW - Cell Membrane

KW - Cell Survival

KW - Cells, Cultured

KW - Dose-Response Relationship, Drug

KW - Gene Expression Regulation

KW - Homocysteine

KW - Journal Article

KW - Membrane Fluidity

KW - Membrane Glycoproteins

KW - Membrane Potential, Mitochondrial

KW - Mitochondria, Heart

KW - Models, Biological

KW - Myocytes, Cardiac

KW - NADPH Oxidase 2

KW - NADPH Oxidases

KW - Necrosis

KW - Phospholipids

KW - Protein Processing, Post-Translational

KW - Rats

KW - Research Support, Non-U.S. Gov't

KW - S-Adenosylhomocysteine

KW - S-Adenosylmethionine

U2 - https://doi.org/10.1007/s10495-007-0077-5

DO - https://doi.org/10.1007/s10495-007-0077-5

M3 - Article

C2 - 17440815

SN - 1360-8185

VL - 12

SP - 1407

EP - 1418

JO - Apoptosis

JF - Apoptosis

IS - 8

ER -