Phytochemical Analysis and Anti-Angiogenic Activity Determination Using Chorioallontoic Membrane (CAM) Assay of Amaranthus spinosus L. (Kulitis) Leaf Extract

Melissa June  Paderog,; Jeannie Joy  Cabangal,; John Patrick  Casiple,; Diane Anthonette  Dato-on,; Jessica May  Daza,; Christien Marie  Demaisip,; Kyle Joshua  Diaz,; Angela Priyah  Dieron,; Dean Allyssa  Diom,; Klint Jun  Ganancial,; Hazel Mae  Panes,

Phytochemical Analysis and Anti-Angiogenic Activity Determination Using Chorioallontoic Membrane (CAM) Assay of Amaranthus spinosus L. (Kulitis) Leaf Extract

Discipline: Pharmacology

Abstract:

Cancer remains the leading cause of mortality worldwide, with approximately 10 million deaths recorded in 2020, accounting for nearly one in every six deaths. A critical factor in cancer progression is angiogenesis, the process by which new blood vessels form to supply oxygen and nutrients, facilitating tumor growth. This study aims to evaluate the anti-angiogenic activity and conduct phytochemical screening of Amaranthus spinosus (Kulitis) leaf extract. The researchers presuppose that there will be a significant mean difference on the average branching points between the treated group, positive control group and negative control group determined by calculating the number of blood vessels post-48 hours of treatment. In this research, 2500 mcg/mL, 1250 mcg/mL, 625 mcg/mL, 312.5 mcg/mL, 156.25 mcg/mL, and 78.125 mcg/mL was evaluated for safety profile using LD50 analysis with brine shrimp assay. The least lethal concentration was then used to for the determination of the anti-angiogenic activity. Phytochemical screening was also performed to confirm the presence of the active constituents responsible for the anti-angiogenic activity. The results from the experiments concluded that A. spinosus exhibits anti-angiogenic activity with mean SD branching points of 640.0000 at 78.125 mcg/mL. Phytochemical screening reveals the presence of saponins, flavonoids, steroids, and tannins.

References:

Cao, Y., Cao, R., & Hedlund, E. M. (2008). Regulation of tumor angiogenesis and metastasis by FGF and PDGF signaling pathways. Journal of Molecular Medicine, 76, 785–789. https://doi.org/10.1007/s00109-008-0337-z
Dada, O. A., Imade, F., & Anifowose, E. M. (2017). Growth and proximate composition of Amaranthus cruentus L. on poor soil amended with compost and arbuscular mycorrhiza fungi. International Journal of Recycling of Organic Waste in Agriculture, 6(3), 195–202. https://doi.org/10.1007/s40093-017-0167-5
Hatami, E., PK, B. N., Sikander, M., Dhasmana, A., Chauhan, S. C., Jaggi, M., & Yallapu, M. M. Tannic acid exhibits antiangiogenesis activity in nonsmall-cell lung cancer cells, ACS Omega 7 (2022) 23939–23949. https://doi.org/10.1021/acsomega.2c02727
Jamiołkowska, A., Skwaryło-Bednarz, B., Kowalski, R., Yildirim, I., & Patkowska, E. (2023). Antifungal potency of amaranth leaf extract: An in vitro study. Plants, 12(8), 1723. https://doi.org/10.3390/plants12081723
Majnooni, M. B., Fakhri, S., Ghanadian, S. M., Bahrami, G., Mansouri, K., Iranpanah, A., ... & Mojarrab, M. (2023). Inhibiting angiogenesis by anti-cancer saponins: from phytochemistry to cellular signaling pathways. Metabolites, 13(3), 323. https://doi.org/10.3390/metabo13030323
Naik, M., Pratush, B., & Dixit, M. (2018). A cost-effective and efficient chick ex-ovo CAM assay protocol to assess angiogenesis. Methods and Protocols, 1(2), 19. https://doi.org/10.3390/mps1020019
Rahman, H., Bukht, T. F. N., Ahmad, R., Almadhor, A., & Javed, A. R. (2023). Efficient breast cancer diagnosis from complex mammographic images using deep convolutional neural network. Computational Intelligence and Neuroscience, 2023(1). https://doi.org/10.1155/2023/7717712
Singh, A., Lama, R., Panda, A., Marndi, S., Devi, R. S., & Kumar, S. (2022). Phytochemical and antibacterial activity of Toddalia asiatica: A wild nutraceutical. Asian Pacific Journal of Health Science, 9(2), 235–239.
Sung, H., Ferlay, J., Siegel, R. L., et al. (2021). Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: A Cancer Journal for Clinicians, 71, 209–249. https://doi.org/10.3322/caac.21660.
Yadav, R., Singh, A. V., Joshi, S., & Kumar, M. (2015). Antifungal and enzyme activity of endophytic fungi isolated from Ocimum sanctum and Aloe vera. African Journal of Microbiology Research, 9(29), 1783–1788. https://doi.org/10.5897/AJMR2015.7451