Cardioprotective Effects of Granulocyte Colony-Stimulating Factor in Swine With Chronic Myocardial Ischemia
Hiroshi Hasegawa, MD, PhD,
Hiroyuki Takano, MD, PhD,
Koji Iwanaga, DVM,
Masashi Ohtsuka, MD, PhD,
Yingjie Qin, MD,
Yuriko Niitsuma, MD,
Kazutaka Ueda, MD,
Tomohiko Toyoda, MD, PhD,
Hiroyuki Tadokoro, MD, and
Issei Komuro, MD, PhD*
Department of Cardiovascular Science and Medicine, Chiba University Graduate School of Medicine, Chiba, Japan
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Figure 2 Fibrosis area of ischemic myocardium. Ischemic areas of control group (A) and granulocyte colony-stimulating factor (G-CSF)-treated group (B) were stained by van Gieson. Bar indicates 50 µm. (C) The average of fibrotic area. Results are given as mean ± SEM (n = 10 each group). *p < 0.05.
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Figure 3 Number of vessels. Endothelial cells of ischemic area of control group (A) and granulocyte colony-stimulating factor (G-CSF)-treated group (B) were identified immunohistochemically using anti-von Willebrand factor antibody. Bar indicates 50 µm. (C) The number of vessels. Results are given as mean ± SEM (n = 10 each group). *p < 0.05.
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Figure 4 Apoptosis of ischemic myocardium. Apoptotic cells of ischemic area at eight weeks after the operation were identified by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-digoxigenin nick end labeling (TUNEL) staining. (A) A representative photograph of TUNEL-positive cells. Bar indicates 30 µm. (B) The percentage of the number of apoptotic cells in total cells. (C) Representative photographs of von Willebrand factor (vWF)- and TUNEL-positive cells. Bar indicates 50 µm. (D) The percentage of vWF- and TUNEL-positive cells to total TUNEL-positive cells. Results are given as mean ± SEM. *p < 0.05.
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Figure 5 Akt1 activity and vascular endothelial growth factor (VEGF) expression in myocardium. Whole tissue lysates were extracted from myocardium of sham-operated swine and ischemic myocardium of operated swine, and subjected to Western blot analysis. (A) The membranes were incubated with antibodies against Akt1 and phospho-Akt1. Results are representative of five independent experiments. (B) Quantitative analysis of Akt1 activity. Results are given as mean ± SEM. (C) A representative photograph of VEGF expression. Results are representative of three independent experiments. (D) Quantitative analysis of VEGF expression. Results are given as mean ± SEM. *p < 0.05.
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) and 40 µg/kg/min (40