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(Circulation. 2001;104:2344.)
© 2001 American Heart Association, Inc.

Basic Science Reports

Therapeutic Angiogenesis Induced by Human Hepatocyte Growth Factor Gene in Rat Diabetic Hind Limb Ischemia Model

Molecular Mechanisms of Delayed Angiogenesis in Diabetes

Yoshiaki Taniyama, MD; Ryuichi Morishita, MD PhD; Kazuya Hiraoka, MD; Motokuni Aoki, MD PhD; Hironori Nakagami, MD; Keita Yamasaki, MD; Kunio Matsumoto, PhD; Toshikazu Nakamura, PhD; Yasufumi Kaneda, MD PhD; Toshio Ogihara, MD PhD

From the Department of Geriatric Medicine (Y.T., R.M., K.H., M.A., H.N., K.Y., T.O.), the Division of Gene Therapy Science (R.M., Y.K.), and the Division of Biochemistry, Department of Oncology, Biomedical Research Center (K.M., T.N.), Osaka University Medical School, Suita, Japan.

Correspondence to Ryuichi Morishita, MD, PhD, Associate Professor, Division of Gene Therapy Science, Osaka University Medical School, 2-2 Yamada-oka, Suita 565, Japan. E-mail morishit{at}geriat.med.osaka-u.ac.jp

Background— Because no study has documented the angiogenic properties of hepatocyte growth factor (HGF) in a diabetes model, we examined the feasibility of gene therapy using HGF to treat peripheral arterial disease in diabetes.

Methods and Results— Because intramuscular injection of luciferase plasmid by the hemagglutinating virus of Japan (HVJ)–liposome method had much higher efficiency than injection of naked plasmid, we used the HVJ-liposome method to transfect the human HGF gene into the rat diabetic hindlimb model. As expected, transfection of human HGF vector resulted in a significant increase in blood flow as assessed by laser Doppler imaging and capillary density, even in the diabetes model, accompanied by the detection of human HGF protein. Interestingly, the degree of natural recovery of blood flow was significantly greater in nondiabetic rats than in diabetic rats. Thus, in an in vitro culture system, we further studied the molecular mechanisms of how diabetes delayed angiogenesis. Importantly, high–D-glucose treatment of endothelial cells resulted in a significant decrease in matrix metalloproteinase (MMP)-1 protein and ets-1 expression in human aortic endothelial cells. Similarly, high D-glucose significantly decreased mRNA and protein of HGF in endothelial cells. Downregulation of MMP-1 and ets-1 by high D-glucose might be due to a significant decrease in HGF, because HGF stimulated MMP-1 production and activated ets-1.

Conclusions— Overall, intramuscular injection of human HGF plasmid induced therapeutic angiogenesis in a rat diabetic ischemic hindlimb model as a potential therapy for peripheral arterial disease. The delay of angiogenesis in diabetes might be due to downregulation of MMP-1 and ets-1 through a decrease in HGF by high D-glucose.

Key Words: peripheral vascular disease • endothelium • angiogenesis • gene therapy • growth substances

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