In vitro screening for angiostatic potential of herbal chemicals.
OBJECTIVE
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.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Cao
Liming
L
Department of Ophthalmology and Visual Sciences, The Chinese University of Hong Kong, Hong Kong.
Liu
Huanming
H
Lam
Dennis Shun-Chiu
DS
Yam
Gary Hin-Fai
GH
Pang
Chi-Pui
CP
D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D061067:Antibodies, Monoclonal, Humanized; D000068258:Bevacizumab; D015153:Blotting, Western; D002392:Catechin / Q000031:analogs & derivatives* / Q000494:pharmacology; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival; D002478:Cells, Cultured; D047188:Chalcones / Q000494:pharmacology*; D004353:Drug Evaluation, Preclinical; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D005136:Eye Proteins / Q000378:metabolism; D051416:Focal Adhesion Protein-Tyrosine Kinases / Q000378:metabolism; D006801:Humans; D009414:Nerve Growth Factors / Q000378:metabolism; D028321:Plant Preparations / Q000494:pharmacology*; D000077185:Resveratrol; D055213:Retinal Pigment Epithelium / Q000166:cytology / Q000187:drug effects; D015843:Serpins / Q000378:metabolism; D013267:Stilbenes / Q000494:pharmacology*; D014471:Umbilical Veins / Q000166:cytology; D042461:Vascular Endothelial Growth Factor A / Q000494:pharmacology; D044004:Xanthones / Q000633:toxicity; D000099174:Pigment Epithelium-Derived Factor
D020533:Angiogenesis Inhibitors; D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D000068258:Bevacizumab; D002392:Catechin; D047188:Chalcones; D005136:Eye Proteins; D051416:Focal Adhesion Protein-Tyrosine Kinases; D009414:Nerve Growth Factors; D028321:Plant Preparations; D000077185:Resveratrol; D015843:Serpins; D013267:Stilbenes; D042461:Vascular Endothelial Growth Factor A; D044004:Xanthones; D000099174:Pigment Epithelium-Derived Factor; C052659:gambogic acid; C040920:isoliquiritigenin; C045651:epigallocatechin gallate