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Anti-angiogenic activity of Gracilaria coronopifolia J.G. Agardh extract by lowering the levels of trace metals (iron, zinc and copper) in duck chorioallantoic membrane and in vitro activation of AMP-kinase

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Abstract

AMP-activated protein kinase (AMPK) is an intracellular energy sensor important in metabolic regulation, cell growth, and survival. However, the specific role of AMPK signaling pathway in the inhibition of angiogenesis remains unclear. The study highlights the activity on AMP activated protein kinase signaling pathways of a marine algae, Gracilaria coronopifolia, and its effects on angiogenesis. It was found that the most potent extract, GCD, inhibited angiogenesis significantly in the duck chorioallantoic membrane assay and also activated the enzyme AMP-kinase, in vitro. The dichloromethane extract was found most active in inhibiting angiogenesis in the duck chorioallantoic membrane (IC50 = 1.21 μg/mL) followed by GCH (IC50 = 3.08 μg/mL) (p = 0.479) and GCM (IC50 = 8.93 μg/mL) (p = 0.042). Benferroni post hoc analysis revealed that there was no significant difference between the percent inhibitions of GCH and GCM extracts (p = 0.479). Consequently, angiogenic inhibition caused lowering of iron, zinc, and copper levels in the duck CAM. Thin layer chromatography and gas chromatography–mass spectrometry revealed the components of each extracts. Notably, this is the first report on the kinase activity of a red algae G. coronopifolia extracts and a colorimetric-based quantification of angiogenesis based on metal content of CAM. Our data also suggest a novel therapeutic approach for inhibiting angiogenesis through the AMPK pathway.

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Acknowledgement

This work was funded by Philippine Department of Science and Technology—Science Education Institute (DOST-SEI). The experiments were done at the University of Santo Tomas Research Center for Natural and Applied Science (UST-RCNAS) and Chung Yuan Christian University (CYCU).

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  1. Oliver B. Villaflores and Katrin Mae M. Ortega contributed equally in preparing the manuscript.

Authors and Affiliations

  1. The Graduate School, University of Santo Tomas, Manila, Philippines

    Oliver B. Villaflores, Katrin Mae M. Ortega, Analin Empaynado-Porto, Dharmatov Rahula Albano & Mary Jho-Anne T. Corpuz

  2. Department of Chemistry, Chung Yuan Christian University, 200 Chung Pei Road, Chung-Li, 320, Taiwan

    Stephen Lirio & Hwa-Kwang Yak

  3. Research Center for the Natural and Applied Sciences, University of Santo Tomas, Manila, Philippines

    Oliver B. Villaflores, Dharmatov Rahula Albano & Mary Jho-Anne T. Corpuz

  4. Department of Biochemistry, Faculty of Pharmacy, University of Santo Tomas, Manila, Philippines

    Oliver B. Villaflores & Dharmatov Rahula Albano

  5. Faculty of Pharmacy, University of Santo Tomas, Manila, Philippines

    Katrin Mae M. Ortega, Analin Empaynado-Porto & Mary Jho-Anne T. Corpuz

  6. Our Lady of Fatima University, Valenzuela, Philippines

    Katrin Mae M. Ortega

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Villaflores, O.B., Ortega, K.M.M., Empaynado-Porto, A. et al. Anti-angiogenic activity of Gracilaria coronopifolia J.G. Agardh extract by lowering the levels of trace metals (iron, zinc and copper) in duck chorioallantoic membrane and in vitro activation of AMP-kinase. Mol Biol Rep 46, 4151–4160 (2019). https://doi.org/10.1007/s11033-019-04864-x

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