Reorganization of cytoskeletal proteins and prolonged life expectancy caused by hepatocyte growth factor in a hamster model of late-phase dilated cardiomyopathy

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Reorganization of cytoskeletal proteins and prolonged life expectancy caused by hepatocyte growth factor in a hamster model of late-phase dilated cardiomyopathy Haruhiko Kondoh, MD, Yoshiki Sawa, MD, Norihide Fukushima, MD, Goro Matsumiya, MD, Shigeru Miyagawa, MD, Satoru Kitagawa- Sakakida, MD, Imran A. Memon, MD, Naomasa Kawaguchi, MD, Nariaki Matsuura, MD, Hikaru Matsuda, MD The Journal of Thoracic and Cardiovascular Surgery Volume 130, Issue 2, Pages 295-302 (August 2005) DOI: 10.1016/j.jtcvs.2004.11.001 Copyright © 2005 The American Association for Thoracic Surgery Terms and Conditions

Figure 1 Detection of human hepatocyte growth factor (HGF) by enzyme-linked immunosorbent assay in transfected hearts. Human HGF protein was detected in hearts 1 week after transfection using the hemagglutinating virus of Japan (HVJ)-liposome method. The HGF expression, however, was transient; only marginal levels of the protein were detected 3 weeks after the transfection. (N = 3 for each group, *P < .05.) The Journal of Thoracic and Cardiovascular Surgery 2005 130, 295-302DOI: (10.1016/j.jtcvs.2004.11.001) Copyright © 2005 The American Association for Thoracic Surgery Terms and Conditions

Figure 2 Cardiac performance. A and B, Left ventricular end-diastolic dimension (Dd) and left ventricular end-systolic dimension (Ds) measured by echocardiography. C, Fractional shortening (FS) calculated using the Dd and Ds. Both the Dd and Ds were gradually enlarged, and FS was decreased in hamsters of the C group. However, hamsters in the H group showed a slightly smaller Dd, significantly smaller Ds, and higher FS during the first 3 weeks after the operation compared with these values in the C group. The effect of HGF transfection was transient, and the Ds began to increase and FS began to decrease by 4 weeks after the transfection. H group (solid boxes); C group (open boxes). (*P < .05 vs before operation [27 weeks old]; #P < .05 vs C group.) The Journal of Thoracic and Cardiovascular Surgery 2005 130, 295-302DOI: (10.1016/j.jtcvs.2004.11.001) Copyright © 2005 The American Association for Thoracic Surgery Terms and Conditions

Figure 3 Suppression of fibrotic change in the left ventricle of TO-2 hamsters by HGF. Tissue sections were stained by hematoxylin-eosin (H&E) or Masson-trichrome dye. The fibrous area was quantified by image analysis in tissue sections stained with Masson-trichrome. The percentage of fibrosis in the H group was significantly suppressed compared with that in the C group. (N = 5 for each group, *P < .05.) The Journal of Thoracic and Cardiovascular Surgery 2005 130, 295-302DOI: (10.1016/j.jtcvs.2004.11.001) Copyright © 2005 The American Association for Thoracic Surgery Terms and Conditions

Figure 4 Immunostaining for alpha-dystroglycan and alpha- and beta-sarcoglycans in cardiomyocytes. A, FIB hamster, 27 weeks old; B, C-group hamster 2 weeks after the operation; C, H-group hamster 2 weeks after the operation; D, H-group hamster, 8 weeks after the operation. Alpha-dystroglycan, and alpha- and beta- sarcoglycans were clearly detected in the sarcolemma of the F1B strain hamsters (A). In the C group, neither dystroglycan nor sarcoglycans was detected (B). In contrast, in the H group, cardiomyocytes expressed alpha-dystroglycan, and alpha- and beta-sarcoglycans 2 weeks after the transfection (C), but this expression disappeared by 8 weeks after the transfection (D). The Journal of Thoracic and Cardiovascular Surgery 2005 130, 295-302DOI: (10.1016/j.jtcvs.2004.11.001) Copyright © 2005 The American Association for Thoracic Surgery Terms and Conditions

Figure 5 Quantitative results of the immunohistologic signals shown in Figure 4. A, Alpha-dystroglycan. B, Alpha-sarcoglycan. C, Beta-sarcoglycan. Although the signals for all 3 proteins had not reached the normal levels observed in F1B mice 2 weeks after the operation, they were significantly higher in the H group than in the C group. (H-2w, 2 weeks after the operation in the H group; H-8w, 8 weeks after the operation in the H group; C-2w, 2 weeks after the operation in the C group; N = 5 for H-2w and C-2w, N = 3 for H-8w, *P < .05.) The Journal of Thoracic and Cardiovascular Surgery 2005 130, 295-302DOI: (10.1016/j.jtcvs.2004.11.001) Copyright © 2005 The American Association for Thoracic Surgery Terms and Conditions

Figure 6 Survivals. The percentage of surviving animals at 40 weeks was 84.2% for the H group, 43.8% for the C group, and 47.9% for the S group. There was no significant difference between the C and S groups, but there was a significant difference between the H and C groups. (H group, N = 19; C group, N = 16; S group, N = 18.) The Journal of Thoracic and Cardiovascular Surgery 2005 130, 295-302DOI: (10.1016/j.jtcvs.2004.11.001) Copyright © 2005 The American Association for Thoracic Surgery Terms and Conditions