Tags

Type your tag names separated by a space and hit enter

Homocysteine-induced cardiomyocyte apoptosis and plasma membrane flip-flop are independent of S-adenosylhomocysteine: a crucial role for nuclear p47(phox).
Mol Cell Biochem. 2011 Dec; 358(1-2):229-39.MC

Abstract

We previously found that homocysteine (Hcy) induced plasma membrane flip-flop, apoptosis, and necrosis in cardiomyocytes. Inactivation of flippase by Hcy induced membrane flip-flop, while apoptosis was induced via a NOX2-dependent mechanism. It has been suggested that S-adenosylhomocysteine (SAH) is the main causative factor in hyperhomocysteinemia (HHC)-induced pathogenesis of cardiovascular disease. Therefore, we evaluated whether the observed cytotoxic effect of Hcy in cardiomyocytes is SAH dependent. Rat cardiomyoblasts (H9c2 cells) were treated under different conditions: (1) non-treated control (1.5 nM intracellular SAH with 2.8 μM extracellular L -Hcy), (2) incubation with 50 μM adenosine-2,3-dialdehyde (ADA resulting in 83.5 nM intracellular SAH, and 1.6 μM extracellular L -Hcy), (3) incubation with 2.5 mM D, L -Hcy (resulting in 68 nM intracellular SAH and 1513 μM extracellular L -Hcy) with or without 10 μM reactive oxygen species (ROS)-inhibitor apocynin, and (4) incubation with 100 nM, 10 μM, and 100 μM SAH. We then determined the effect on annexin V/propodium iodide positivity, flippase activity, caspase-3 activity, intracellular NOX2 and p47(phox) expression and localization, and nuclear ROS production. In contrast to Hcy, ADA did not induce apoptosis, necrosis, or membrane flip-flop. Remarkably, both ADA and Hcy induced a significant increase in nuclear NOX2 expression. However, in contrast to ADA, Hcy additionally induced nuclear p47(phox) expression, increased nuclear ROS production, and inactivated flippase. Incubation with SAH did not have an effect on cell viability, nor on flippase activity, nor on nuclear NOX2-, p47phox expression or nuclear ROS production. HHC-induced membrane flip-flop and apoptosis in cardiomyocytes is due to increased Hcy levels and not primarily related to increased intracellular SAH, which plays a crucial role in nuclear p47(phox) translocation and subsequent ROS production.

Authors+Show Affiliations

Department of Pathology, VU University Medical Center, De Boelelaan 1117, 1007 MB Amsterdam, The Netherlands.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

21739151

Citation

Sipkens, Jessica A., et al. "Homocysteine-induced Cardiomyocyte Apoptosis and Plasma Membrane Flip-flop Are Independent of S-adenosylhomocysteine: a Crucial Role for Nuclear P47(phox)." Molecular and Cellular Biochemistry, vol. 358, no. 1-2, 2011, pp. 229-39.
Sipkens JA, Krijnen PA, Hahn NE, et al. Homocysteine-induced cardiomyocyte apoptosis and plasma membrane flip-flop are independent of S-adenosylhomocysteine: a crucial role for nuclear p47(phox). Mol Cell Biochem. 2011;358(1-2):229-39.
Sipkens, J. A., Krijnen, P. A., Hahn, N. E., Wassink, M., Meischl, C., Smith, D. E., Musters, R. J., Stehouwer, C. D., Rauwerda, J. A., van Hinsbergh, V. W., & Niessen, H. W. (2011). Homocysteine-induced cardiomyocyte apoptosis and plasma membrane flip-flop are independent of S-adenosylhomocysteine: a crucial role for nuclear p47(phox). Molecular and Cellular Biochemistry, 358(1-2), 229-39. https://doi.org/10.1007/s11010-011-0973-4
Sipkens JA, et al. Homocysteine-induced Cardiomyocyte Apoptosis and Plasma Membrane Flip-flop Are Independent of S-adenosylhomocysteine: a Crucial Role for Nuclear P47(phox). Mol Cell Biochem. 2011;358(1-2):229-39. PubMed PMID: 21739151.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Homocysteine-induced cardiomyocyte apoptosis and plasma membrane flip-flop are independent of S-adenosylhomocysteine: a crucial role for nuclear p47(phox). AU - Sipkens,Jessica A, AU - Krijnen,Paul A J, AU - Hahn,Nynke E, AU - Wassink,Melissa, AU - Meischl,Christof, AU - Smith,Desirée E C, AU - Musters,René J P, AU - Stehouwer,Coen D A, AU - Rauwerda,Jan A, AU - van Hinsbergh,Victor W M, AU - Niessen,Hans W M, Y1 - 2011/07/08/ PY - 2011/01/13/received PY - 2011/06/28/accepted PY - 2011/7/9/entrez PY - 2011/7/9/pubmed PY - 2012/2/22/medline SP - 229 EP - 39 JF - Molecular and cellular biochemistry JO - Mol. Cell. Biochem. VL - 358 IS - 1-2 N2 - We previously found that homocysteine (Hcy) induced plasma membrane flip-flop, apoptosis, and necrosis in cardiomyocytes. Inactivation of flippase by Hcy induced membrane flip-flop, while apoptosis was induced via a NOX2-dependent mechanism. It has been suggested that S-adenosylhomocysteine (SAH) is the main causative factor in hyperhomocysteinemia (HHC)-induced pathogenesis of cardiovascular disease. Therefore, we evaluated whether the observed cytotoxic effect of Hcy in cardiomyocytes is SAH dependent. Rat cardiomyoblasts (H9c2 cells) were treated under different conditions: (1) non-treated control (1.5 nM intracellular SAH with 2.8 μM extracellular L -Hcy), (2) incubation with 50 μM adenosine-2,3-dialdehyde (ADA resulting in 83.5 nM intracellular SAH, and 1.6 μM extracellular L -Hcy), (3) incubation with 2.5 mM D, L -Hcy (resulting in 68 nM intracellular SAH and 1513 μM extracellular L -Hcy) with or without 10 μM reactive oxygen species (ROS)-inhibitor apocynin, and (4) incubation with 100 nM, 10 μM, and 100 μM SAH. We then determined the effect on annexin V/propodium iodide positivity, flippase activity, caspase-3 activity, intracellular NOX2 and p47(phox) expression and localization, and nuclear ROS production. In contrast to Hcy, ADA did not induce apoptosis, necrosis, or membrane flip-flop. Remarkably, both ADA and Hcy induced a significant increase in nuclear NOX2 expression. However, in contrast to ADA, Hcy additionally induced nuclear p47(phox) expression, increased nuclear ROS production, and inactivated flippase. Incubation with SAH did not have an effect on cell viability, nor on flippase activity, nor on nuclear NOX2-, p47phox expression or nuclear ROS production. HHC-induced membrane flip-flop and apoptosis in cardiomyocytes is due to increased Hcy levels and not primarily related to increased intracellular SAH, which plays a crucial role in nuclear p47(phox) translocation and subsequent ROS production. SN - 1573-4919 UR - https://www.unboundmedicine.com/medline/citation/21739151/Homocysteine_induced_cardiomyocyte_apoptosis_and_plasma_membrane_flip_flop_are_independent_of_S_adenosylhomocysteine:_a_crucial_role_for_nuclear_p47_phox__ L2 - https://doi.org/10.1007/s11010-011-0973-4 DB - PRIME DP - Unbound Medicine ER -