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Abstract

Hypoxic response element (HRE) offers satisfactory control over expression of hVEGF₁₆₅ in cell levels. However, the characteristics of regenerated blood vessels induced by long-term expression of transferred hVEGF₁₆₅ under control of HRE in vivo remain unknown. This study aims to investigate the effect of HRE on control of long-term expression of rAAV-delivered hVEGF₁₆₅ gene to ischemic myocardium and evaluate characteristics of angiogenesis induced by hVEGF₁₆₅ in vivo. Rabbit ischemic heart models were established surgically, rAAV-9HRE-hVEGF₁₆₅ was transfected to ischemia hearts subjected to 12 week ischemia. Molecular biological and immunohistochemistry were employed to determine expressions of HIF-1α and hVEGF₁₆₅. Microvessel densities of CD31⁺ and α-SMA⁺ regenerated vessels were also evaluated. Expressions of both hVEGF₁₆₅ mRNA and protein were upregulated following over-expression of endogenous HIF-1α early after ischemia, peaked at 4–6 weeks post-MI, declined, and approached pre-ischemia level at the end of 12 weeks of ischemia (P < 0.01). The significantly upregulated CD31 in hVEGF₁₆₅-treated hearts presented from 8 to 12 weeks of ischemia compared with the control (P < 0.01). However, α-SMA expression was rapidly downregulated after ischemia and remained lower than the control level by the end of 12 weeks post-MI (P < 0.01). Overexpression of hVEGF₁₆₅ controlled by HIF-1α-HRE system shows a stably regional angiogenic efficacy in vivo. But, VEGF, as an early angiogenic cytokine, is inadequate for mediating histologically mature vessels in ischemia myocardium.

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Angiogenesis Induced by h VEGF 165 Gene Controlled by Hypoxic Response Elements in Rabbit Ischemia Myocardium

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