Korean Circ J.  2008 Sep;38(9):455-461. 10.4070/kcj.2008.38.9.455.

The Expression of Cardiac Ankyrin Repeat Protein in an Animal Model of Adriamycin-Induced Cardiomyopathy

• Chung WB
• Youn HJ
• Choi YS
• Park CS
• Oh YS
• Chung WS
• Kim JH
• Lee EH

• KMID: 1776345
• DOI: http://doi.org/10.4070/kcj.2008.38.9.455

Abstract

BACKGROUND AND OBJECTIVES
Cardiac ankyrin repeat protein (CARP) is an embryonic nuclear protein, and its expression is increased under conditions of pressure or volume overload and also in the failing heart. Adriamycin is a cardiotoxic chemotherapeutic agent, and it suppresses the expression of CARP. We compared the expressions of CARP in the myocardium of normotensive rats and spontaneously hypertensive rats (SHRs) that suffered with adriamycin-induced cardiomyopathy. MATERIALS AND METHODS: 36 Wistar-Kyoto rats (WKYs) and 36 SHRs were divided into the adriamycin-administered and saline-administered groups. Adriamycin (2.5 mg/kg) and saline were injected intraperitoneally twice a week for 3 weeks. All the animals were sacrificed 3 weeks after the last injection. Immunohistochemical staining was performed on the left ventricles with using synthesized polyclonal CARP antibody. The positively stained areas were measured by using an image analysis program, and the CARP volume fractions (CaVF) were calculated. RESULTS: CARP was diffusely expressed in the cytoplasm of the myocytes in all the groups. The number of CARP expressing cells was increased in the SHRs. The CaVFs was 5.96+/-5.11% in the WKYs and it was 9.04+/-6.26% in the SHRs (p=0.014). The CaVF was 2.26+/-4.74% in the adriamycin-administered WKYs and it was 1.24+/-4.32% in the adriamycin-administered SHRs (p=0.32). The adriamycin-administered WKYs and SHRs showed significantly decreased CARP expressions, as compared to the saline-administered groups (p<0.001 and p<0.001, respectively). CONCLUSION: These results suggest that CARP is closely related to the pathogenesis of adriamycin-induced cardiomyopathy and it probably plays a pivotal role for the adriamycin cardiac toxicity observed in hypertensive rats.

Keyword

Ankyrin, repeat; Hypertension; Adriamycin; Cardiomyopathies

MeSH Terms

Animals
Ankyrin Repeat
Ankyrins
Cardiomyopathies
Carps
Cytoplasm
Doxorubicin
Heart
Heart Ventricles
Hypertension
Models, Animal
Muscle Cells
Myocardium
Nuclear Proteins
Rats
Rats, Inbred SHR
Ankyrins
Doxorubicin
Nuclear Proteins

Figure

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  Fig. 1 Immunohistochemical staining of the cultured 293 T cells. The scale bar indicates 25 um. The brown colored cells (black arrow) indicate that the CARP antibody effectively detects the CARP expressing cells. CARP: cardiac ankyrin repeat protein.

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  Fig. 2 Western blot analysis of the CARP expression in the 293 T cells. The CARP antibody was shown to specifically recognize CARP. CARP: cardiac ankyrin repeat protein.

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  Fig. 3 Schedule of the adriamycin injections and normal saline injections and the echocardiography. Echo: echocardiography, BP: blood pressure, i.p.: intraperitoneally.

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  Fig. 4 Measurement of the CARP expression (WKY-NS). A: the red colored regions represent the CARP positive areas according to the image analysis program (Image Pro-plus 4.5). The scale bar indicates 100 um. B: magnified image of a black rectangle in A. The scale bar indicates 25 um. CARP: cardiac ankyrin repeat protein, WKY-NS: normal saline treated Wistar-Kyoto rat.

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  Fig. 5 Immunohistochemical staining of the myocardium. The scale bars indicate 100 um. The brown colored cells are the CARP expressing cells that were detected by the CARP antibody. A: normal saline-treated WKY myocardium. B: adriamycin treated WKY myocardium. C: normal saline treated SHR myocardium. D: adriamycin treated SHR myocardium. CARP: cardiac ankyrin repeat protein, WKY: Wistar-Kyoto rat, SHR: spontaneously hypertensive rat.

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  Fig. 6 Immunohistochemical staining of the myocardium. The scale bars indicate 25 um. The brown colored cells are the CARP expressing cells that were detected by the CARP antibody. A: normal saline treated WKY myocardium. B: adriamycin treated WKY myocardium. C: normal saline treated SHR myocardium. D: adriamycin treated SHR myocardium. CARP: cardiac ankyrin repeat protein, WKY: Wistar-Kyoto rat, SHR: spontaneously hypertensive rat.

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  Fig. 7 Comparisons of the CARP volume fraction (CaVF). WKY-NS: normal saline treated Wistar-Kyoto rat, SHR-NS: normal saline treated spontaneously hypertensive rat, WKY-AD: adriamycin treated Wistar-Kyoto rat, SHR-AD: adriamycin treated spontaneously hypertensive rat.

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