RESUMO
ETHNOPHARMACOLOGICAL RELEVANCE: Diabetic foot ulceration results in high risk of lower extremity amputation, and represents a significant health care expenditure worldwide. Radix Astragali (RA) and Radix Rehmanniae (RR) are widely used Chinese medicinal herbs in treating diabetes, and have shown positive effects in enhancing wound healing in diabetic foot ulcer animal model. MATERIALS AND METHODS: The angiogenic efficacy of NF3, a simplified 2-herb formula consisting of RA and RR in 2:1 ratio, was investigated. Median lethal concentration (LC50) and median effective concentration (EC50) were determined by treating zebrafish embryos with different concentrations of NF3 from 20 hpf to 72 hpf. The angiogenic activity of NF3 was examined in zebrafish embryos in vivo and by rat aortic ring assay in vitro. Cell cycle analysis of endothelial cells induced by NF3 was analyzed by flow cytometry using transgenic zebrafish Tg(fli1:EGFP). Real-time PCR was used to analyze mRNA expression profiles of selected genes involved in VEGF, FGF and MAPK pathways. RESULTS: NF3 enhanced blood vessel formation as indicated by extra growth of intersegmental vessels in zebrafish embryos, and increased microvessels formation in rat aortic ring. NF3 also enhanced endothelial cells proliferation as shown by increased percentage of cells accumulating in S phase and G2/M phase of the cell cycle. NF3 exposure significantly induced up-regulation of VEGF-A, Flk-1, fgf1 and bRaf expression in zebrafish embryos. CONCLUSIONS: Our results demonstrated that NF3 was effective in promoting angiogenesis in zebrafish embryos and by rat aortic ring assay, which provided scientific basis to support the use of NF3 as potential therapeutics in treating diabetic foot ulceration.
Assuntos
Indutores da Angiogênese/farmacologia , Aorta/efeitos dos fármacos , Medicamentos de Ervas Chinesas/farmacologia , Células Endoteliais/efeitos dos fármacos , Neovascularização Fisiológica/efeitos dos fármacos , Indutores da Angiogênese/uso terapêutico , Indutores da Angiogênese/toxicidade , Proteínas Angiogênicas/genética , Animais , Animais Geneticamente Modificados , Proliferação de Células/efeitos dos fármacos , Pé Diabético/tratamento farmacológico , Relação Dose-Resposta a Droga , Medicamentos de Ervas Chinesas/uso terapêutico , Medicamentos de Ervas Chinesas/toxicidade , Embrião não Mamífero/irrigação sanguínea , Embrião não Mamífero/efeitos dos fármacos , Células Endoteliais/metabolismo , Pontos de Checagem da Fase G2 do Ciclo Celular/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Humanos , Dose Letal Mediana , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Sistema de Sinalização das MAP Quinases/genética , Masculino , Neovascularização Fisiológica/genética , Proteína Proto-Oncogênica c-fli-1/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Reação em Cadeia da Polimerase em Tempo Real , Pontos de Checagem da Fase S do Ciclo Celular/efeitos dos fármacos , Peixe-Zebra/embriologia , Peixe-Zebra/genéticaRESUMO
Chinese materia medica (CMM) is becoming increasingly important in modern health care, with the potential for new or improved clinical protocols and reduction in treatment costs. Conventional approaches to drug discovery are based on knowledge of biological systems and screen phenotypes in the context of a whole organism. It will be valuable to identify the CMM that would induce certain biological responses (such as angiogenesis). The authors have developed a database that they plan to commercialize that contains traditional knowledge of Chinese medicine and pharmacology along with their own experimental data from controlled scientific observations by using the zebrafish as a model of CMM-induced pathology. The database is visualized and functions via the World Wide Web by subscription or license. The authors have also written software for personal digital assistant (PDA) devices that supports multiple users performing screening experiments worldwide. This provides a platform for the study of CMM, and data mining of this resource will help evaluate CMM in the context of experimental observations of biological aberrations.
