EVALUATION OF TAXIFOLIN AND PHLORETIN AS ANTIANGIOGENIC: AN IN VIVO, IN VITRO EXPERIMENTAL ANALYSIS
Keywords:
Angiogenesis, Flavonoids, Chick Chorio Allantoic Membrane (CAM), Dorsal Skinfold Chamber (DSFC), Vascular Endothelial Growth Factor (VEGF)Abstract
Objective: To examine the antiangiogenic effect of some flavonoids (taxifolin and phloretin) against various In vivo and In vitro experimental models such as Chick Chorio Allantoic Membrane (CAM) assay, Dorsal Skinfold Chamber (DSFC) and Tube formation assay.
Methods:Three different concentrations of drug solution (40µM, 80 µM, 120 µM) and (50 µM, 100 µM, 150 µM) were applied topically during CAM and DSFC assay. These assays were performed to detect antiangiogenic effect of the selected molecules. In tube formation assay, vascular endothelial growth factor stimulated Human Umbelical Vain Endothelial Cells (HUVECs).
Results:Antiangiogenic results showed that different concentration of taxifolin and phloretin inhibited new blood vessels and vessels branches per area of CAMs. Similarly, there are reduced numbers and diameter of blood vessels when the doses of the drug increased which are observed in different experimental groups in DSFC model. In vitro anti angiogenic effect of our molecules was further confirmed by inhibition of tube formation on matrigel matrix that involved HUVECs. All the test groups were compared with the control group.
Conclusion: The data reported here provide evidence that flavonoids taxifolin and phloretin revealed prominent anti angiogenic activities.
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Folkman J. Angiogenesis in cancer, vascular, rheumatoid and other disease. Nat Med 1995;1:27-31.
Risau W, Feri K, Joe M. Mechanism of angiogenesis. Nature 1997;386:671-4.
Auerbach R, Rachel L, Brenda S, Louis K, Nasim A. Angiogenesis assays a critical overview. Clin Chem 2003;49:132−40.
Stern CD. The chick a great model system becomes even greater. Dev Cell 2005;8:9-17.
Liu M, Scanlon CS, Banerjee R, Russo N, Inglehart RC. The histone methyltransferase EZH2 mediates tumor progression on the chick chorioallantoic membrane assay, a novel model of head and neck squamous cell carcinoma. Transl Oncol 2013;6:273-81.
Murphy JB. Transplant ability of tissues to the embryo of foreign species It’s bearing on questions of tissue specificity and tumor immunity. J Exp Med 1913;17:482-93.
Zijlstra A, Mellor R, Panzarella G, Aimes RT, Hooper JD. A quantitative analysis of rate-limiting steps in the metastatic cascade using human-specific real-time polymerase chain reaction. Cancer Res 2002;62:7083-92.
Algire GH. An adaptation of the transparent chamber technique to the mouse. J Natl Cancer Inst 1943;4:1–11.
Algire GH, Legallais FY. Recent developments in the transparent chamber technique as adapted to the mouse. J Natl Cancer Inst 1949;10:225-53.
Papenfuss HD, Gross JF, Intaglietta M, Treese FA. A transparent access chamber for the rat dorsal skinfold. Microvasc Res 1979;18:311-8.
Kurth EF, Chan FL. Dihydroquercetin as an antioxidant. J Am Oil Chem Soc 1951;28:433–6.
Luo H, Jiang BH, King SM, Chen YC. Inhibition of cell growth and VEGF expression in ovarian cancer cells by flavonoids. Nutr Cancer 2008;60:800–9.
Brusselmans K, Vrolix R, Verhoeven G, Swinnen JV. Induction of cancer cell apoptosis by flavonoids is associated with their ability to inhibit fatty acid synthase activity. J Biol Chem 2005;280:5636–45.
Furuta E, Pai SK, Zhan R, Bandyopadhyay S, Watabe M, Mo YY, et al. Fatty acid synthase gene is up-regulated by hypoxia via activation of Akt and sterol regulatory element binding protein-1. Cancer Res 2008;68:1003–11.
Plísková M, Vondrácek J, Kren V, Gazák R, Sedmera P, Walterová D, et al. Effects of silymarin flavonolignans and synthetic silybin derivatives on estrogen and aryl hydrocarbon receptor activation. Toxicology 2005;215:80–9.
Nam NH, Kim Y, You YJ, Hong DH, Kim HM, Ahn BZ. Cytotoxic 2,5 dihydroxychalcones with unexpected antiangiogenic activity. Eur J Med Chem 2003;38:179–87.
Lee YS, Lim SS, Shin KH, Kim YS, Ohuchi K, Jung SH. Anti-angiogenic and anti-tumor activities of 2-hydroxy-4-methoxychalcone. Biol Pharm Bull 2006;29:1028–31.
Park K, Lim D, Park SD, Kim MY, Kim Y. Angiogenesis inhibitor derived from angiostatin active sites. Bull Korean Chem Soc 2004;25:1331–5.
Soucy NV, Ihnat MA, Kamat CD, Hess L, Mark JP, Klei LR, et al. Arsenic stimulates angiogenesis and tumorigenesis in vivo. Toxicol Sci 2003;76:271–9.
Rajesh N, Gacche, Harshala D, Shegokar, Dhananjay S, Gond, et al. Evaluation of selected flavonoids as antiangiogenic, Anticancer, and radical scavenging agents an experimental and in silico analysis. Cell Biochem Biophys 2011;61:651–63.
Moy AJ, White SM, Elmer SI, Justin L, Nudelman MJ, Costantini SJ. Wide field functional imaging of blood flow and hemoglobin oxygen saturation in the rodent dorsal window chamber. Microvasc Res 2011;82:199-09.
Ashton AW, Yokota R, John G, Zhaoi S, Suadicanii SO. Inhibition of endothelial cell migration, intercellular communication, and vascular tube formation by thromboxane A2. J Biol Chem 1999;274:35562–70.
Yeh JC, Cindrova-Davies T, Belleri M, Morbidelli L, Miller N, Cho CW, et al. The Natural compound n-butylidenephthalide derived from the volatile oil of radix angelica sinensis inhibits angiogenesis in vitro and in vivo. Angiogenesis 2011;14:187–97.
Brooks PC, Montgomery AM, Cheresh DA. Use of the 10-day-old chick embryo model for studying angiogenesis. Methods Mol Biol 1999;129:257–69.
Nijveldt RJ, Nood E, Hoorn DEC, Boelens PG, Norren K, Leeuwen PAM. Flavonoids a review of probable mechanisms of action and potential applications. Am J Clin Nutr 2001;74:418–25.
Kim MH. Flavonoids inhibit VEGF/bFGF-induced angiogenesis In vitro by inhibiting the matrix-degrading proteases. J Cell Biochem 2003;89:529–38.
Yuki H, Kiyoshi E, Yoko Y, Setsuko K, Yasuaki H, Yoshiteru I, et al. Specific inhibition of hypoxiainducible factor (HIF) 1a activation and vascular endothelial growth factor (VEGF) production by flavonoids. Biol Pharm Bull 2003;26:1379–83.
Mojzis J, Varinska L, Mojzisova G, Kostova I, Mirossay L. Antiangiogenic effects of flavonoids and chalcones. Pharmacol Res 2008;57:259–65.
Yoshida M, Sakai T, Hosokawa N, Marui N, Matsumoto K, Fujioka A, et al. The effect of quercetin of cell cycle progression and growth of human gastric cancer cells. FEBS Lett 1990;260:10–3.
Thamilarashi AN, Mangalagowri A, Winkins Santosh. Antiangiogenic activity of natural flavonoid myricetin on chick chorioallantoic membrane (CAM) in vivo. Int J Pharm 2014;4:160-5.
Friedlander M, Brooks PC, Shaffer RW, Kincaid CM, Varner JA, Cheresh DA. Definition of two angiogenic pathways by distinct alpha v integrins. Science 1995;270:1500-2.
Kerbel RS. A cancer therapy resistant to resistance. Nature 1997;390:404-7.
Dai ZH, Lu WF, Gao J, Kang HF, Ma YG, Zhang SQ, et al. Anti-angiogenic effect of the total flavonoids in Scutellaria barbata D Don. BMC Complementary Altern Med 2013;13:150-60.
Mignatti P, Rifkin DB. Plasminogen activators and matrix metalloproteinases in angiogenesis. Enzyme Protein 1996;49:117–37.
Bischoff J. Cell adhesion and angiogenesis. J Clin Invest 1997;99:373–6.
Ferrara N, Gerber HP, LeCouter J. The biology of VEGF and its receptors. J Nat Med 2003;9:669–76.
Lamalice L, Le Boeuf F, Huot J. Endothelial cell migration during angiogenesis. Circ Res 2007;100:782–94.
Nishi M, Abe Y, Tomii Y, Tsukamoto H, Kijima H, Yamazaki H, et al. Cell binding isoforms of vascular endothelial growth factor-A (VEGF189) contribute to blood flow-distant metastasis of pulmonary adenocarcinoma. Int J Oncol 2005;26:1517–24.
Brogi E, Schatteman G, Wu T, Kim EA, Varticovski L, Keyt B, et al. Hypoxia-induced paracrine regulation of vascular endothelial growth factor receptor expression. J Clin Invest 1996;97:469–76.