stilbenes and isoliquiritigenin

Herbal medicine has long been used in traditional medicinal systems. The authors carried out a first-line screening of four herbal chemicals with reported antioxidative properties and capabilities to suppress endothelial cell growth and migration. These herbal chemicals were isoliquiritigenin (ISL) from licorice, epigallocatechin gallate (EGCG) from green tea, resveratrol (Rst) from grapes, and gambogic acid (GA) from the resin of Garcinia hanburyi.. Cytotoxicity was studied by MTT cell viability/proliferation assay on human retinal pigment epithelial cells (ARPE19). Effects on vascular endothelial growth factor (VEGF)-induced endothelial cell proliferation and migration were investigated by a scratch-wound model using human umbilical vein endothelial cells (HUVECs). The effects on VEGF signaling in HUVECs were analyzed by Western blotting.. At sub-cytotoxic levels, ISL (10 μM), EGCG (50 μM), and Rst (10 μM) suppressed HUVEC proliferation and migration under VEGF (20 ng/mL) stimulation in our scratch-wound model. HUVEC migration was reduced more by ISL and EGCG than bevacizumab, a humanized monoclonal antibody against VEGF. The efficiency of Rst was similar to that of bevacizumab. GA, however, was toxic to cells even at nanomolar concentrations. Western blot analysis showed that these chemicals affected focal adhesion kinase activation and expression of pigment epithelial growth factor.. ISL, EGCG, and Rst are highly effective and efficient in suppressing endothelial cell proliferation and migration, with low cytotoxicity on ARPE19 and HUVEC lines. They are potentially useful for further investigation to develop antiangiogenic therapies by virtue of their small molecular sizes for easy penetration through tissue cells and their low effective dosages.

Topics: Angiogenesis Inhibitors; Antibodies, Monoclonal; Antibodies, Monoclonal, Humanized; Bevacizumab; Blotting, Western; Catechin; Cell Movement; Cell Proliferation; Cell Survival; Cells, Cultured; Chalcones; Drug Evaluation, Preclinical; Endothelium, Vascular; Enzyme-Linked Immunosorbent Assay; Eye Proteins; Focal Adhesion Protein-Tyrosine Kinases; Humans; Nerve Growth Factors; Plant Preparations; Resveratrol; Retinal Pigment Epithelium; Serpins; Stilbenes; Umbilical Veins; Vascular Endothelial Growth Factor A; Xanthones

Stilbene (STS) and chalcone (CHS) synthases are homodimeric, related plant-specific polyketide synthases. Both perform a sequential condensation of three acetate units to a starter residue to form a tetraketide intermediate that is folded to the ring systems specific to the different products. Protein cross-linking and site-directed mutagenesis identified a subunit contact site in position 158, close to the active site (Cys169). This suggested that the active site pockets may be neighboring, possibly alternating in the condensing reactions rather than acting independently. This was investigated by coexpression of active site mutants with differently mutated, inactive proteins. With both STS and CHS, the heterodimers synthesized the end products, indicating that each subunit performed all three condensations. In co-action with a monomeric reductase, CHS also synthesizes 6'-deoxychalcone, but with the chalcone as second product when using plant preparations. The two enzymes expressed as single species in Escherichia coli synthesized both products, and both were also obtained with a CHS heterodimer containing a single active site. The results showed that 6'-deoxychalcone synthesis required no other plant factor and that the formation of two products may be an intrinsic property of the interaction between dimeric CHS and monomeric reductase.

Topics: Acyltransferases; Amino Acid Sequence; Binding Sites; Chalcone; Chalcones; Genetic Complementation Test; Molecular Sequence Data; Plants; Stilbenes