Anti-angiogenic effects of Astaxanthin in human umbilical vein Endothelial cells induced by vascular endothelial growth factor

Angiogenesis is the development of new blood vessels from preexisting ones that is essential for tumour growth and metastasis. Anti-VEGF therapy such as Bevacizumab; Avastin has been used as anti-angiogenic agent in cancer treatment yet the treatment is frequently associated with severe adverse effects. Alternatives are investigated for complementary natural products with comparable anti-angiogenic effectiveness and minimal adverse effects. Despite research demonstrating that astaxanthin (ATX) possesses anti-oxidant and anti-inflammatory properties, its effect on the angiogenesis process is yet unknown. Therefore, the purpose of this study is to ascertain ATX's anti-angiogenic properties. By using in-vitro (cytotoxicity, proliferation, migration, and tube-formation assays) and in-silico (network pharmacology and molecular docking) studies, the effects of ATX on VEGF-induced human umbilical vein endothelial cells (HUVEC) angiogenesis were assessed. Concentrations used for cell cytotoxicity assay were ranging from 50 – 3.125 µM while for cell proliferation, cell migration and tube formation assays used concentrations ranging from 25 – 6.25 µM. The angiogenesis-inhibiting compound Suramin served as the positive control. The results revealed that ATX exhibited cytotoxic effect (IC50 = 75.09 µM) by statistically significant reduced cell viability (%) of HUVECs in concentration-dependent manner (P<0.05) and at 50µM ATX killed 33% of the HUVEC when treated for 24 h. Furthermore, ATX demonstrated an anti-proliferative effect, leading to a statistically significant reduction in cell viability (%) of VEGF-induced HUVECs in a concentration-dependent manner (P < 0.05). This effect was observed after a 72-hour treatment, using a 4-hour baseline for comparison. Next, ATX exhibited anti-migratory effect by statistically inhibited migration in concentration-dependent manner (P<0.05) through scratch wound healing assay and the migration rate (%) at 25 µM is 20% for 8 h. Moreover, ATX inhibited tube formation by statistically significant decreased the average of tube length (µm) in concentration-dependent manner (P<0.05) through tube formation assay for 6 h. To further elucidate the potential molecular mechanisms underlying these effects, we transitioned to in-silico studies. Four common hub genes: RXR, CCND1, CDK1, and HDAC1 were discovered through network pharmacology which were deemed to be key target genes of ATX against angiogenesis. Furthermore, ATX may exert its effects through serine/threonine/tyrosine kinase activity, modulating inflammatory responses in the cytoplasm, and interacting with pathways involved in cancer. Besides, the interactions of proteins and ligands of four hub genes (RXRA, CCND1, CDK1, and HDAC1) and VEGFR2, FGFR1, and Tie-2 were determined through molecular docking. The results revealed that ATX exhibits strong binding affinity to all seven proteins, with a significantly higher affinity observed for VEGFR2 compared to the others. In conclusion, ATX demonstrated anti-angiogenic effects on VEGF-induced HUVECs through cytotoxicity, proliferation, migration, and tube-formation assays, with potential molecular mechanisms predicted through in silico studies. This study provides useful basis for further investigations on ATX against angiogenesis related diseases.

Text FPSK (m) 2024 17 - Declaration Form.pdf Restricted to Repository staff only Available under License Creative Commons Attribution Non-commercial No Derivatives. Download (534kB) Text FPSK (m) 2024 17 - Full Text.pdf Available under License Creative Commons Attribution Non-commercial No Derivatives. Download (4MB) Text FPSK (m) 2024 17.pdf Restricted to Repository staff only Available under License Creative Commons Attribution Non-commercial No Derivatives. Download (4MB)

Actions (login required)

View Item