Cilostazol Promotes Angiogenesis and Increases Cell Proliferation After Myocardial Ischemia–Reperfusion Injury Through a cAMP-Dependent Mechanism
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Previous study indicated the protective role of cilostazol in ischemia–reperfusion (I/R) injury. Here, we aimed to explore the function of cilostazol in myocardial I/R injury and the underlying mechanism.
The myocardial I/R injury rat model was constructed, and the expression levels of vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), basic fibroblast growth factor (bFGF), platelet-derived growth factor receptor b (PDGF-B) and the number of new blood vessels were measured by qRT-PCR and immunohistochemistry. VSMC and HUVEC cells were treated with hypoxia to induce in vivo I/R injury model. The protein expression of AKT, endothelial nitric oxide synthase (eNOS) and apoptosis-related protein levels were detected by western blotting. Besides, the positive staining rate and cell viability were tested by 5-bromo-2-deoxyuridine (Brdu)/4′,6-diamidino-2-phenylindole (DAPI) or DAPI/TdT-mediated dUTP Nick-End Labeling (TUNEL) staining and MTT assay.
Cilostazol promoted angiogenesis by increasing the number of new blood vessels and up-regulating the expression of VEGF, HGF, bFGF and PDGF-B in myocardial I/R-injury rat model. The in vitro experiments showed that cilostazol increased the level of eNOS and AKT, and also enhanced cell proliferation in hypoxia-treated VSMC and HUVEC cells. Furthermore, after 8-Br-cAMP treatment, VEGF, HGF, bFGF, PDGF-B, p-AKT and p-eNOS expression were up-regulated, while cleaved-caspase 3 and cleaved-PARP expression were down-regulated. In addition, the effects of cilostazol on cell viability and apoptosis were aggravated by 8-Br-cAMP and attenuated after KT-5720 treatment.
Cilostazol could promote angiogenesis, increase cell viability and inhibit cell apoptosis, consequently protecting myocardial tissues against I/R-injury through activating cAMP.
KeywordsCilostazol Ischemia–reperfusion injury Myocardium CAMP Angiogenesis
The authors declare that they have no competing interests, and all authors should confirm its accuracy.
- 4.Biscetti, F., G. Pecorini, G. Straface, V. Arena, E. Stigliano, S. Rutella, F. Locatelli, F. Angelini, G. Ghirlanda, and A. Flex. Cilostazol promotes angiogenesis after peripheral ischemia through a VEGF-dependent mechanism. Int. J. Cardiol. 167(3):910–916, 2013. https://doi.org/10.1016/j.ijcard.2012.03.103.CrossRefGoogle Scholar
- 6.Chao, T. H., S. Y. Tseng, Y. H. Li, P. Y. Liu, C. L. Cho, G. Y. Shi, H. L. Wu, and J. H. Chen. A novel vasculo-angiogenic effect of cilostazol mediated by cross-talk between multiple signalling pathways including the ERK/p38 MAPK signalling transduction cascade. Clin. Sci. (Lond.) 123(3):147–159, 2012. https://doi.org/10.1042/cs20110432.CrossRefGoogle Scholar
- 7.Choi, H. I., D. Y. Kim, S. J. Choi, C. Y. Shin, S. T. Hwang, K. H. Kim, and O. Kwon. The effect of cilostazol, a phosphodiesterase 3 (PDE3) inhibitor, on human hair growth with the dual promoting mechanisms. J. Dermatol. Sci. 91(1):60–68, 2018. https://doi.org/10.1016/j.jdermsci.2018.04.005.CrossRefGoogle Scholar
- 11.Hayward, C. P., K. A. Moffat, J. F. Castilloux, Y. Liu, J. Seecharan, S. Tasneem, S. Carlino, A. Cormier, and G. E. Rivard. Simultaneous measurement of adenosine triphosphate release and aggregation potentiates human platelet aggregation responses for some subjects, including persons with Quebec platelet disorder. Thromb. Haemost. 107(4):726–734, 2012. https://doi.org/10.1160/th11-10-0740.CrossRefGoogle Scholar
- 12.Herath, S. C., T. Lion, M. Klein, D. Stenger, C. Scheuer, J. H. Holstein, P. Morsdorf, M. F. Rollmann, T. Pohlemann, M. D. Menger, et al. Stimulation of angiogenesis by cilostazol accelerates fracture healing in mice. J. Orthop. Res. 33(12):1880–1887, 2015. https://doi.org/10.1002/jor.22967.CrossRefGoogle Scholar
- 13.Hol, P. K., P. S. Lingaas, R. Lundblad, K. A. Rein, K. Vatne, H. J. Smith, S. Nitter-Hauge, and E. Fosse. Intraoperative angiography leads to graft revision in coronary artery bypass surgery. Ann. Thorac. Surg. 78(2):502–505, 2004; discussion 505. https://doi.org/10.1016/j.athoracsur.2004.03.004.CrossRefGoogle Scholar
- 15.Huang, Y., Y. Cheng, J. Wu, Y. Li, E. Xu, Z. Hong, Z. Li, W. Zhang, M. Ding, X. Gao, et al. Cilostazol as an alternative to aspirin after ischaemic stroke: a randomised, double-blind, pilot study. Lancet Neurol. 7(6):494–499, 2008. https://doi.org/10.1016/s1474-4422(08)70094-2.CrossRefGoogle Scholar
- 16.Huang, J. H., X. H. Huang, Z. Y. Chen, Q. S. Zheng, and R. Y. Sun. Equivalent dose conversion between animals and between animals and humans in pharmacological tests (In Chinese). Chin. J. Clin. Pharmacol. Ther. 9(9):1069–1072, 2004.Google Scholar
- 17.Jeon, C., S. C. Candia, J. C. Wang, E. M. Holper, M. Ammerer, R. E. Kuntz, and L. Mauri. Relative spatial distributions of coronary artery bypass graft insertion and acute thrombosis: a model for protection from acute myocardial infarction. Am. Heart J. 160(1):195–201, 2010. https://doi.org/10.1016/j.ahj.2010.04.004.CrossRefGoogle Scholar
- 19.Li, H., D. H. Hong, Y. K. Son, S. H. Na, W. K. Jung, Y. M. Bae, E. Y. Seo, S. J. Kim, I. W. Choi, and W. S. Park. Cilostazol induces vasodilation through the activation of Ca(2+)-activated K(+) channels in aortic smooth muscle. Vascul. Pharmacol. 70:15–22, 2015. https://doi.org/10.1016/j.vph.2015.01.002.CrossRefGoogle Scholar
- 21.Liu, Y., T. Wang, J. Yan, N. Jiagbogu, D. A. Heideman, A. E. Canfield, and M. Y. Alexander. HGF/c-Met signalling promotes Notch3 activation and human vascular smooth muscle cell osteogenic differentiation in vitro. Atherosclerosis 219(2):440–447, 2011. https://doi.org/10.1016/j.atherosclerosis.2011.08.033.CrossRefGoogle Scholar
- 22.Matsui, Y., H. Takagi, X. Qu, M. Abdellatif, H. Sakoda, T. Asano, B. Levine, and J. Sadoshima. Distinct roles of autophagy in the heart during ischemia and reperfusion: roles of AMP-activated protein kinase and Beclin 1 in mediating autophagy. Circ. Res. 100(6):914–922, 2007. https://doi.org/10.1161/01.res.0000261924.76669.36.CrossRefGoogle Scholar
- 25.Oyama, N., Y. Yagita, M. Kawamura, Y. Sugiyama, Y. Terasaki, E. Omura-Matsuoka, T. Sasaki, and K. Kitagawa. Cilostazol, not aspirin, reduces ischemic brain injury via endothelial protection in spontaneously hypertensive rats. Stroke 42(9):2571–2577, 2011. https://doi.org/10.1161/strokeaha.110.609834.CrossRefGoogle Scholar
- 26.Peters, T. H., V. Sharma, E. Yilmaz, W. J. Mooi, A. J. Bogers, and H. S. Sharma. DNA microarray and quantitative analysis reveal enhanced myocardial VEGF expression with stunted angiogenesis in human tetralogy of Fallot. Cell Biochem. Biophys. 67(2):305–316, 2013. https://doi.org/10.1007/s12013-013-9710-9.CrossRefGoogle Scholar
- 29.Rosen, E. M., K. Lamszus, J. Laterra, P. J. Polverini, J. S. Rubin, and I. D. Goldberg. HGF/SF in angiogenesis. Ciba Found. Symp. 212:215–226; discussion 227–219, 1997.Google Scholar
- 30.Sanada, F., Y. Kanbara, Y. Taniyama, R. Otsu, M. Carracedo, Y. Ikeda-Iwabu, J. Muratsu, K. Sugimoto, K. Yamamoto, H. Rakugi, et al. Induction of angiogenesis by a type III phosphodiesterase inhibitor, cilostazol, through activation of peroxisome proliferator-activated receptor-gamma and cAMP pathways in vascular cells. Arterioscler. Thromb. Vasc. Biol. 36(3):545–552, 2016. https://doi.org/10.1161/atvbaha.115.307011.CrossRefGoogle Scholar
- 31.Shimizu, T., T. Osumi, K. Niimi, and K. Nakagawa. Physico-chemical properties and stability of cilostazol. Arzneimittelforschung 35(7a):1117–1123, 1985.Google Scholar
- 33.von Heesen, M., S. Muller, U. Keppler, M. J. Strowitzki, C. Scheuer, M. K. Schilling, M. D. Menger, and M. R. Moussavian. Preconditioning by cilostazol protects against cold hepatic ischemia-reperfusion injury. Ann Transplant. 20:160–168, 2015. https://doi.org/10.12659/aot.893031.CrossRefGoogle Scholar
- 35.Wang, C., C. Wang, Q. Liu, Q. Meng, J. Cang, H. Sun, J. Peng, X. Ma, X. Huo, and K. Liu. Aspirin and probenecid inhibit organic anion transporter 3-mediated renal uptake of cilostazol and probenecid induces metabolism of cilostazol in the rat. Drug Metab. Dispos. 42(6):996–1007, 2014. https://doi.org/10.1124/dmd.113.055194.CrossRefGoogle Scholar
- 39.Zhang, G. G., X. Teng, Y. Liu, Y. Cai, Y. B. Zhou, X. H. Duan, J. Q. Song, Y. Shi, C. S. Tang, X. H. Yin, et al. Inhibition of endoplasm reticulum stress by ghrelin protects against ischemia/reperfusion injury in rat heart. Peptides 30(6):1109–1116, 2009. https://doi.org/10.1016/j.peptides.2009.03.024.CrossRefGoogle Scholar