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MicroRNAs Are Involved in Homocysteine-Induced Cardiac Remodeling

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Abstract

Elevated level of homocysteine (Hcy) called hyperhomocysteinemia (HHcy) is one of the major risk factors for chronic heart failure. Although the role of Hcy in cardiac remodeling is documented, the regulatory mechanism involved therein is still nebulous. MicroRNAs (miRNAs) and dicer have been implicated in regulation of cardiovascular diseases. Dicer is the only known enzyme involved in miRNA maturation. We investigated the involvement of dicer and miRNA in Hcy-induced cardiac remodeling. HL-1 cardiomyocytes were cultured in different doses of Hcy. Total RNA was isolated and RT-PCR and real-time PCR was performed for dicer, MMP-2,-9, TIMP-1,-3, and NOX-4. MiRNA microarray was used for analyzing the differential expression of miRNAs. Individual miRNA assay was also done. Western blotting was used to assess the MMP-9 expression in HHcy cardiomyocytes. The RT-PCR results suggest that dicer expression is enhanced in HHcy cardiomyocytes suggesting its involvement in cardiac remodeling caused due to high dose of Hcy. On the other hand, high dose of Hcy increased NOX-4 expression, a marker for oxidative stress. Additionally, HHcy cardiomyocytes showed elevated levels of MMP-2,-9 and TIMP-1,-3, and reduced expression of TIMP-4, suggesting cardiac remodeling due to oxidative stress. The miRNA microarray assay revealed differential expression of 11 miRNAs and among them miR-188 show dramatic downregulation. These findings suggest that dicer and miRNAs especially miR-188 are involved in Hcy-induced cardiac remodeling.

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Abbreviations

Hcy :

Homocysteine

HHcy:

Hyperhomocysteinemia

MMP:

Matrix metalloproteinase

TIMP:

Tissue inhibitor of metalloproteinase

NOX:

Nicotinamide adenine diphosphate oxidase

CHF:

Congestive heart failure

CVD:

cardiovascular diseases

miR/miRNA:

microRNA

RT-PCR:

Reverse transcription polymerase chain reaction

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Acknowledgment

A part of the study was supported by NIH grants HL 71010, HL-74185, and HL-88012.

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Authors and Affiliations

  1. Department of Physiology & Biophysics, University of Louisville, School of Medicine, A-1215, 500 South Preston Street, Louisville, KY, 40202, USA

    Paras K. Mishra, Neetu Tyagi, Soumi Kundu & Suresh C. Tyagi

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  1. Paras K. Mishra
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  2. Neetu Tyagi
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  3. Soumi Kundu
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  4. Suresh C. Tyagi
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Correspondence to Suresh C. Tyagi.

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Mishra, P.K., Tyagi, N., Kundu, S. et al. MicroRNAs Are Involved in Homocysteine-Induced Cardiac Remodeling. Cell Biochem Biophys 55, 153–162 (2009). https://doi.org/10.1007/s12013-009-9063-6

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  • DOI: https://doi.org/10.1007/s12013-009-9063-6

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