Asian Journal of Pharmaceutical Research and Health Care

Open Access Open Access  Restricted Access Subscription Access

Comparative Experimental Studies of Few L-Type and T-Type Ca2+ Channel Blockers against In-Ovo and In-Vitro Models of Angiogenesis


Affiliations
1 Department of Pharmacology and Toxicology, Northern Border University, Rafha, Saudi Arabia
2 Department of Pharmaceutical Chemistry, Northern Border University, Rafha, Saudi Arabia
 

Angiogenesis is the development of new blood vessels from pre-existing one. The ion-channels on endothelium plays vital role in cell proliferation and related angiogenesis. We aimed to investigate the effects of L-type (Verapamil and Diltiazem) and T-type (Ethosuximide) Calcium Channel Blockers (CCBs) on neovascularization. The effects on neovascularization were investigated by in-ovo (CAM, Chick Chorioallantoic membrane) and in-vitro (aortic ring assay) methods. Each test drug was tested for at least 3 doses and the anti-angiogenic effect was compared with Suramin as standard and normal control groups. Various vital parameters were recorded during the experiment like the number of blood capillaries, sprouts formation, angiogenic score etc. The L-type Ca2+ channel blockers Verapamil at the dose of 50μM, 110 and 220 μM/disk has shown significant (p < 0.001) reduction in the number of branching points in CAM assay. For the further confirmation, angiogenic activity was evaluated in vitro by rat aortic ring assay method; the area of sprouts was reduced by the medium and high dose of verapamil. Diltiazem has demonstrated modest anti-angiogenic activity by both the models, whereas T-type calcium channel blocker ethosuximide has not shown any effect on the neovascularization. Among all the tested drugs verapamil has shown the promising anti-angiogenic property. Thus verapamil and diltiazem may have antiangiogenic activity defines novel beneficial effects in angiogenic mediated pathological conditions in addition to their main indications in cardiovascular complications.

Keywords

Angiogenesis, CAM Assay, L-Type Ca2+ Channel Blockers, Rat Aortic Ring Assay, T-Type Calcium Channel Blocker.
User
Notifications
Font Size

  • Folkman J, Klagsbrun M. Angiogenic factors. Science. 1987 Jan 23; 235(4787):442–7. crossref PMid:2432664

  • Folkman J. Seminars in medicine of the beth israel hospital, Boston. Clinical applications of research on angiogenesis. N Engl J Med. 1995 Dec 28; 333(26):1757–63. crossref PMid:7491141

  • Otrock ZK, Mahfouz RA, Makarem JA, Shamseddine AI. Understanding the biology of angiogenesis: Review of the most important molecular mechanisms. Blood Cells Mol Dis. 2007 Sep-Oct; 39(2):212–20. crossref PMid:17553709

  • Tahergorabi Z, Khazaei M. A review on angiogenesis and its assays. Iran J Basic Med Sci. 2012 Nov; 15(6):1110–26. PMid:23653839 PMCid:PMC3646220

  • Deryugina EI, Quigley JP. Tumor angiogenesis: MMP-mediated induction of intravasation- and metastasis-sustaining neovasculature. Matrix Biol. 2015 May-Jul; 44-46:94–112. crossref PMid:25912949 PMCid:PMC5079283

  • D’Amore PA, Thompson RW. Mechanisms of angiogenesis. Annu Rev Physiol. 1987; 49:453–64. crossref PMid:2436570

  • Augustin HG. Antiangiogenic tumour therapy: will it work? Trends Pharmacol Sci. 1998 Jun; 19(6):216–22. crossref

  • Balk SD. Calcium as a regulator of the proliferation of normal, but not of transformed, chicken fibroblasts in a plasma-containing medium. Proc Natl Acad Sci U S A. 1971 Feb; 68(2):271–5. crossref PMid:5277067 PMCid:PMC388915

  • Boynton AL, Whitfield JF, Isaacs RJ, Morton HJ. Control of 3T3 cell proliferation by calcium. In Vitro. 1974 Jul-Aug; 10:12–7. crossref PMid:4471173

  • Durham AC, Walton JM. Calcium ions and the control of proliferation in normal and cancer cells. Biosci Rep. 1982 Jan; 2(1):15–30. crossref PMid:7037065

  • Boynton AL, Whitfield JF. Different calcium requirements for proliferation of conditionally and unconditionally tumorigenic mouse cells. Proc Natl Acad Sci U S A. 1976 May; 73(5):1651–4. crossref PMid:1064038 PMCid:PMC430357

  • Boynton AL, Whitfield JF, Isaacs RJ, Tremblay RG. Different extracellular calcium requirements for proliferation of nonneoplastic, preneoplastic, and neoplastic mouse cells. Cancer Res. 1977 Aug; 37(8 Pt 1):2657–61. PMid:872093

  • Parsons PG. Selective proliferation of human tumour cells in calcium-depleted medium. Aust J ExpBiol Med Sci. 1978 Jun; 56(3):297–300. crossref PMid:101190

  • Paul D, Ristow HJ. Cell cycle control by Ca++-ions in mouse 3T3 cells and in transformed 3T3 cells. J Cell Physiol. 1979 Jan; 98(1):31-9. crossref PMid:762200

  • Lokman NA, Elder AS, Ricciardelli C, Oehler MK. Chick Chorioallantoic Membrane (CAM) assay as an in vivo model to study the effect of newly identified molecules on ovarian cancer invasion and metastasis. Int J Mol Sci. 2012; 13(8):9959–70. crossref PMid:22949841 PMCid:PMC3431839

  • Burgermeister J1, Paper DH, Vogl H, Linhardt RJ, Franz G. LaPSvS1, a (1-->3)-beta-galactan sulfate and its effect on angiogenesis in vivo and in vitro. Carbohydr Res. 2002 Sep 9; 337(16):1459–66. crossref

  • Krenn L, Paper DH. Inhibition of angiogenesis and inflammation by an extract of red clover (Trifoliumpratense L.). Phytomedicine. 2009 Dec; 16(12):1083–8. crossref PMid:19665361

  • AlMalki WH, Shahid I, Mehdi AY, Hafeez MH. Assessment methods for angiogenesis and current approaches for its quantification. Indian J Pharmacol. 2014 May-Jun; 46(3):251–6. crossref PMid:24987169 PMCid:PMC4071699

  • Fernandes G, Barone A, Dziak R. Effects of verapamil on bone cancer cells In Vitro. J Cell Biol Cell Metab. 2016; 3:013.

  • Caglar Y, Ali Cetin, Demirci F, Zubeyde AP, Tuba K, Ahmet A, Meral C, Ozlem K Y, Ismihan G. Anti-angiogenic effects of diltiazem, imatinib, and bevacizumab in the CAM assay. International Journal of Scientific and Research Publications. 2013; 3(8).

  • Gomora JC1, Daud AN, Weiergräber M, Perez-Reyes E. Block of cloned human T-type calcium channels by succinimide antiepileptic drugs. Mol Pharmacol. 2001 Nov; 60(5):1121–32. crossref PMid:11641441

  • Kerbel R, Folkman J. Clinical translation of angiogenesis inhibitors. Nat Rev Cancer. 2002 Oct; 2(10):727–39. crossref PMid:12360276

  • Fidler IJ. The pathogenesis of cancer metastasis: the ‘seed and soil’ hypothesis revisited. Nat Rev Cancer. 2003 Jun; 3(6):453–8. crossref PMid:12778135

  • Bergers G, Benjamin LE. Tumorigenesis and the angiogenic switch. Nat Rev Cancer. 2003 Jun; 3(6):401–10. crossref PMid:12778130

  • Capiod T. Cell proliferation, calcium influx and calcium channels. Biochimie. 2011 Dec; 93(12):2075–9. crossref PMid:21802482

  • Harris-Hooker SA, Gajdusek CM, Wight TN, Schwartz SM. Neovascular responses induced by cultured aortic endothelial cells. J Cell Physiol. 1983 Mar; 114(3):302–10. crossref PMid:6187756

  • Nguyen M, Shing Y, Folkman J. Quantitation of angiogenesis and antiangiogenesis in the chick embryo chorioallantoic membrane. Microvasc Res. 1994 Jan; 47(1):31–40. crossref PMid:7517489

  • Lam CF, Liu YC, Tseng FL, Sung YH, Huang CC, Jiang MJ, Tsai YC. High-dose morphine impairs vascular endothelial function by increased production of superoxide anions. Anesthesiology. 2007 Mar; 106(3):532–7. crossref PMid:17325512

  • Mason RP1, Mak IT, MW Trumbore, Mason PE. Antioxidant properties of calcium antagonists related to membrane biophysical interactions. Am J Cardiol. 1999 Aug 19; 84(4A):16L–22L. PMid:10480441

  • Hayman SR, Leung N, Grande JP, Garovic VD. VEGF inhibition, hypertension, and renal toxicity. Curr Oncol Rep. 2012 Aug; 14(4):285–94. crossref PMid:22544560 PMCid:PMC3746763

  • Nicosia RF, Ottinetti A. Growth of microvessels in serumfree matrix culture of rat aorta. A quantitative assay of angiogenesis in vitro. Lab Invest. 1990 Jul; 63(1):115–22. PMid:1695694

  • Nicosia RF, Villaschi S. Autoregulation of angiogenesis by cells of the vessel wall. Int Rev Cytol. 1999; 185:1–43. crossref

  • Mariot P, Vanoverberghe K, Lalevee N, Rossier MF, Prevarskaya N. Overexpression of an alpha 1H (Cav3.2) T-type calcium channel during neuroendocrine differentiation of human prostate cancer cells. J Biol Chem. 2002 Mar 29; 277(13):10824-33. Epub 2002 Jan 17. crossref PMid:11799114

  • Ciapa B, Pesando D, Wilding M, Whitaker M. Cell-cycle calcium transients driven by cyclic changes in inositol trisphosphate levels. Nature. 1994 Apr 28; 368(6474):875–8. crossref PMid:8159248

  • Choi DW. Ionic dependence of glutamate neurotoxicity. J Neurosci. 1987 Feb; 7(2):369–79. PMid:2880938


Abstract Views: 427

PDF Views: 175




  • Comparative Experimental Studies of Few L-Type and T-Type Ca2+ Channel Blockers against In-Ovo and In-Vitro Models of Angiogenesis

Abstract Views: 427  |  PDF Views: 175

Authors

Vijay R. Chidrawar
Department of Pharmacology and Toxicology, Northern Border University, Rafha, Saudi Arabia
Mohd. Imran
Department of Pharmaceutical Chemistry, Northern Border University, Rafha, Saudi Arabia

Abstract


Angiogenesis is the development of new blood vessels from pre-existing one. The ion-channels on endothelium plays vital role in cell proliferation and related angiogenesis. We aimed to investigate the effects of L-type (Verapamil and Diltiazem) and T-type (Ethosuximide) Calcium Channel Blockers (CCBs) on neovascularization. The effects on neovascularization were investigated by in-ovo (CAM, Chick Chorioallantoic membrane) and in-vitro (aortic ring assay) methods. Each test drug was tested for at least 3 doses and the anti-angiogenic effect was compared with Suramin as standard and normal control groups. Various vital parameters were recorded during the experiment like the number of blood capillaries, sprouts formation, angiogenic score etc. The L-type Ca2+ channel blockers Verapamil at the dose of 50μM, 110 and 220 μM/disk has shown significant (p < 0.001) reduction in the number of branching points in CAM assay. For the further confirmation, angiogenic activity was evaluated in vitro by rat aortic ring assay method; the area of sprouts was reduced by the medium and high dose of verapamil. Diltiazem has demonstrated modest anti-angiogenic activity by both the models, whereas T-type calcium channel blocker ethosuximide has not shown any effect on the neovascularization. Among all the tested drugs verapamil has shown the promising anti-angiogenic property. Thus verapamil and diltiazem may have antiangiogenic activity defines novel beneficial effects in angiogenic mediated pathological conditions in addition to their main indications in cardiovascular complications.

Keywords


Angiogenesis, CAM Assay, L-Type Ca2+ Channel Blockers, Rat Aortic Ring Assay, T-Type Calcium Channel Blocker.

References





DOI: https://doi.org/10.18311/ajprhc%2F2018%2F19587