Effect of 7 Kinds of Active Constituents from Astragalus membranaceus on the Expression of HIF-1
Kuijun Zhao, Peng‐Fei Tu
Abstract
Kuijun Zhao, Peng‐Fei Tu
Abstract
OBJECTIVE: To explore the effect of formononetin,ononin,calycosin-O-β-D-glucoside,calycosin,acetylastragalosideⅠ,astragalosideⅡ, astragaloside Ⅳ from Astragalus membranaceus on the expression of hypoxia-induced factor-1(HIF-1). METHODS: The HRE, derived from EPO containing a highly conserved HIF-1 binding site, were cloned into pBI-GL vectors that contained luciferase reporter genes. The HEK-293T cells were transfected cDNA constructs by calcium phosphate precipitation methods. The activity of luciferase was determined after 48 h treatment. The expression of HIF-1 was evaluated using luciferase report gene system. RESULTS:10 μmol·L-1 formononetin promote expression of HIF-1.Ononin had an effect on the expression of HIF-1 in a concentration-dependent manner. Low concentration could elevate the expression of HIF-1 while high concentration down-regulated the expression. Other 5 monomers had no obvious effect on the expression of HIF-1. CONCLUSION:The study provide reference for the anti-tumor effect of A. membranaceus and therapeutic effect of it for other disease.
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OBJECTIVE: To explore the effect of formononetin,ononin,calycosin-O-β-D-glucoside,calycosin,acetylastragalosideⅠ,astragalosideⅡ, astragaloside Ⅳ from Astragalus membranaceus on the expression of hypoxia-induced factor-1(HIF-1). METHODS: The HRE, derived from EPO containing a highly conserved HIF-1 binding site, were cloned into pBI-GL vectors that contained luciferase reporter genes. The HEK-293T cells were transfected cDNA constructs by calcium phosphate precipitation methods. The activity of luciferase was determined after 48 h treatment. The expression of HIF-1 was evaluated using luciferase report gene system. RESULTS:10 μmol·L-1 formononetin promote expression of HIF-1.Ononin had an effect on the expression of HIF-1 in a concentration-dependent manner. Low concentration could elevate the expression of HIF-1 while high concentration down-regulated the expression. Other 5 monomers had no obvious effect on the expression of HIF-1. CONCLUSION:The study provide reference for the anti-tumor effect of A. membranaceus and therapeutic effect of it for other disease.
Key concepts: Calycosin, Formononetin, Luciferase, Astragaloside, Astragalus, Chemistry, Transfection, Molecular biology