Ticagrelor suppresses oxidized low‑density lipoprotein‑induced endothelial cell apoptosis and alleviates atherosclerosis in ApoE‑/‑ mice via downregulation of PCSK9

Xia,Xiaodong; Li,Jiao; Liang,Xue; Zhang,Shengjie; Liu,Tong; Liu,Jinying; Arif,Muhammad; Li,Guangping

doi:10.3892/mmr.2018.9779

Ticagrelor suppresses oxidized low‑density lipoprotein‑induced endothelial cell apoptosis and alleviates atherosclerosis in ApoE‑/‑ mice via downregulation of PCSK9

Authors:
- Xiaodong Xia
- Jiao Li
- Xue Liang
- Shengjie Zhang
- Tong Liu
- Jinying Liu
- Muhammad Arif
- Guangping Li
View Affiliations

Affiliations: Department of Cardiology, Second Hospital of Tianjin Medical University, Tianjin Key Laboratory of Ionic‑Molecular Function of Cardiovascular Disease, Tianjin Institute of Cardiology, Tianjin 300211, P.R. China, Department of Pharmacy, The Second People's Hospital of Liaocheng, Shandong, Liaocheng 252600, P.R. China, College of Life Science, Nankai University, Tianjin 300071, P.R. China
Published online on: December 20, 2018 https://doi.org/10.3892/mmr.2018.9779
Pages: 1453-1462
Copyright: © Xia et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Although ticagrelor has been demonstrated to possess an anti‑atherosclerosis (AS) effect, its underlying mechanism remains unclear. In the present study, it was investigated whether ticagrelor reduces oxidized low‑density lipoprotein (ox‑LDL)‑induced endothelial cell apoptosis, an initial step for the development of AS, and alleviates AS in apolipoprotein‑E‑deficient (ApoE‑/‑) mice by inhibiting the expression of proprotein convertase subtilisin/kexin type 9 (PCSK9). The human endothelial cell line EAhy926 was treated with ox‑LDL, ox‑LDL + ticagrelor (40 µmol/l) and ox‑LDL + ticagrelor (60 µmol/l) for 24 h. Cell apoptosis was detected using Annexin V‑fluorescein isothiocyanate/propidium iodide staining. The expression levels of PCSK9, apoptosis‑associated proteins and signaling pathways were determined by reverse transcription‑quantitative polymerase chain reaction and western blotting. ApoE‑/‑ mice fed a high‑fat diet were used to induce an AS model. After 20 weeks, ApoE‑/‑ mice were randomly assigned to receive saline or ticagrelor intragastrically for 10 days. The formation of atherosclerotic plaques was detected by hematoxylin and eosin staining. The expression of PCSK9 in the arterial tissues was measured by immunohistochemistry. The results demonstrated that treatment with ticagrelor was able to decrease ox‑LDL‑induced apoptosis in a concentration‑dependent manner (40 µmol/l vs. ox‑LDL, 17.58±2.66 vs. 27.25±5.54%; 60 µmol/l vs. ox‑LDL, 12.26±1.54 vs. 27.25±5.54%). The mRNA and protein expression level of PCSK9 significantly decreased following treatment with ticagrelor, accompanied with upregulation of B‑cell lymphoma (Bcl) 2 and downregulation of Bcl‑2 associated X, apoptosis regulator, caspase‑3, p38, phosphorylated‑(p) p38, p‑c‑Jun N‑terminal kinases (JNK), p‑extracellular signal‑regulated kinases and the ratio of p‑JNK to JNK. Histological analysis of arterial tissues revealed ticagrelor markedly decreased the atherosclerotic plaque area and inhibited the expression of PCSK9. The present results suggested that ticagrelor may alleviate AS via downregulation of PCSK9‑mediated endothelial cell apoptosis, which may be JNK‑dependent.

View Figures

View References

1	Clarke R, Du H, Kurmi O, Parish S, Yang M, Arnold M, Guo Y, Bian Z, Wang L, Chen Y, et al: Burden of carotid artery atherosclerosis in Chinese adults: Implications for future risk of cardiovascular diseases. Eur J Prev Cardiol. 24:647–656. 2017. View Article : Google Scholar : PubMed/NCBI
2	Barquera S, Pedroza-Tobías A, Medina C, Hernández-Barrera L, Bibbins-Domingo K, Lozano R and Moran AE: Global overview of the epidemiology of atherosclerotic cardiovascular disease. Arch Med Res. 46:328–338. 2015. View Article : Google Scholar : PubMed/NCBI
3	Mathers CD and Loncar D: Projections of global mortality and burden of disease from 2002 to 2030. PLoS Med. 3:e4422006. View Article : Google Scholar : PubMed/NCBI
4	Chen M, Masaki T and Sawamura T: LOX-1, the receptor for oxidized low-density lipoprotein identified from endothelial cells: Implications in endothelial dysfunction and atherosclerosis. Pharmacol Ther. 95:89–100. 2002. View Article : Google Scholar : PubMed/NCBI
5	Hong D, Bai YP, Gao HC, Wang X, Li LF, Zhang GG and Hu CP: Ox-LDL induces endothelial cell apoptosis via the LOX-1-dependent endoplasmic reticulum stress pathway. Atherosclerosis. 235:310–317. 2014. View Article : Google Scholar : PubMed/NCBI
6	Bao MH, Zhang YW and Zhou HH: Paeonol suppresses oxidized low-density lipoprotein induced endothelial cell apoptosis via activation of LOX-1/p38MAPK/NF-κB pathway. J Ethnopharmacol. 146:543–551. 2013. View Article : Google Scholar : PubMed/NCBI
7	Xu F, Sun Y, Chen Y, Sun Y, Li R, Liu C, Zhang C, Wang R and Zhang Y: Endothelial cell apoptosis is responsible for the formation of coronary thrombotic atherosclerotic plaques. Tohoku J Exp Med. 218:25–33. 2009. View Article : Google Scholar : PubMed/NCBI
8	Durand E, Scoazec A, Lafont A, Boddaert J, Al Hajzen A, Addad F, Mirshahi M, Desnos M, Tedgui A and Mallat Z: In vivo induction of endothelial apoptosis leads to vessel thrombosis and endothelial denudation a clue to the understanding of the mechanisms of thrombotic plaque erosion. Circulation. 109:2503–2506. 2004. View Article : Google Scholar : PubMed/NCBI
9	Badimon L and Vilahur G: Thrombosis formation on atherosclerotic lesions and plaque rupture. J Intern Med. 276:618–632. 2015. View Article : Google Scholar
10	Raskob GE, Angchaisuksiri P, Blanco AN, Büller H, Gallus A, Hunt BJ, Hylek EM, Kakkar TL, Konstantinides SV, Mccumber M, et al: Thrombosis: A major contributor to global disease burden. Semin Thromb Hemost. 40:724–735. 2014. View Article : Google Scholar : PubMed/NCBI
11	Zhang Y, Mu Q, Zhou Z, Song H, Zhang Y, Wu F, Jiang M, Wang F, Zhang W, Li L, et al: Protective effect of irisin on atherosclerosis via suppressing oxidized low density lipoprotein induced vascular inflammation and endothelial dysfunction. PLoS One. 11:e01580382016. View Article : Google Scholar : PubMed/NCBI
12	Wang L, Hao Q, Wang YD, Wang WJ and Li DJ: Protective effects of dehydroepiandrosterone on atherosclerosis in ovariectomized rabbits via alleviating inflammatory injury in endothelial cells. Atherosclerosis. 214:47–57. 2011. View Article : Google Scholar : PubMed/NCBI
13	Wang GF, Shi CG, Sun MZ, Wang L, Wu SX, Wang HF, Xu ZQ and Chen DM: Tetramethylpyrazine attenuates atherosclerosis development and protects endothelial cells from ox-LDL. Cardiovasc Drugs Ther. 27:199–210. 2013. View Article : Google Scholar : PubMed/NCBI
14	Lee CW, Hwang I, Park CS, Lee H, Park DW, Kang SJ, Lee SW, Kim YH Park SW and Park SJ: Comparison of differential expression of P2Y¹² receptor in culprit coronary plaques in patients with acute myocardial infarction versus stable angina pectoris. Am J Cardiol. 108:799–803. 2011. View Article : Google Scholar : PubMed/NCBI
15	Jeong HS, Hong SJ, Cho SA, Kim JH, Cho JY, Lee SH, Joo HJ, Park JH, Yu CW and Lim DS: Comparison of ticagrelor versus prasugrel for inflammation, vascular function, and circulating endothelial progenitor cells in diabetic patients with non-ST-segment elevation acute coronary syndrome requiring coronary stenting. JACC Cardiovasc Interv. 10:1646–1658. 2017. View Article : Google Scholar : PubMed/NCBI
16	Warnholtz A, Ostad MA, Velich N, Trautmann C, Schinzel R, Walter U and Munzel T: A single loading dose of clopidogrel causes dose-dependent improvement of endothelial dysfunction in patients with stable coronary artery disease: Results of a double-blind, randomized study. Atherosclerosis. 196:689–695. 2008. View Article : Google Scholar : PubMed/NCBI
17	Willoughby SR, Luu LJ, Cameron JD, Nelson AJ, Schultz CD, Worthley SG and Worthley MI: Clopidogrel improves microvascular endothelial function in subjects with stable coronary artery disease. Heart Lung Circ. 23:534–541. 2014. View Article : Google Scholar : PubMed/NCBI
18	Kim HK, Jeong MH, Lim KS, Kim JH, Lim HC, Kim MC, Hong YJ, Kim SS, Park KH and Chang KS: Effects of ticagrelor on neointimal hyperplasia and endothelial function, compared with clopidogrel and prasugrel, in a porcine coronary stent restenosis model. Int J Cardiol. 240:326–331. 2017. View Article : Google Scholar : PubMed/NCBI
19	Preusch MR, Rusnak J, Staudacher K, Mogler C, Uhlmann L, Sievers P, Bea F, Katus HA, Blessing E and Staudacher I: Ticagrelor promotes atherosclerotic plaque stability in a mouse model of advanced atherosclerosis. Drug Des Devel Ther. 10:2691–2699. 2016. View Article : Google Scholar : PubMed/NCBI
20	Ren H, Li ML, Jiang J, Zhang Y, Zhang Y and Zhu X: Effects of clopidogrel on vascular proliferation and apoptosis in an atherosclerotic rabbit model. J Cardiovasc Pharmacol. 55:617–624. 2010. View Article : Google Scholar : PubMed/NCBI
21	Li J, Liang X, Wang Y, Xu Z and Li G: Investigation of highly expressed PCSK9 in atherosclerotic plaques and ox-LDL-induced endothelial cell apoptosis. Mol Med Rep. 16:1817–1825. 2017. View Article : Google Scholar : PubMed/NCBI
22	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
23	Capodanno D, Dharmashankar K and Angiolillo DJ: Mechanism of action and clinical development of ticagrelor, a novel platelet ADP P2Yreceptor antagonist. Expert Rev Cardiovasc Ther. 8:151–158. 2010. View Article : Google Scholar : PubMed/NCBI
24	Wang J, Chen L, Li H, Yang J, Gong Z, Wang B and Zhao X: Clopidogrel reduces apoptosis and promotes proliferation of human vascular endothelial cells induced by palmitic acid via suppression of the long non-coding RNA HIF1A-AS1 in vitro. Mol Cell Biochem. 404:203–210. 2015. View Article : Google Scholar : PubMed/NCBI
25	Sachais BS: Platelet-endothelial interactions in atherosclerosis. Curr Atheroscler Rep. 3:412–416. 2001. View Article : Google Scholar : PubMed/NCBI
26	Nylander S, Femia EA, Scavone M, Berntsson P, Asztély AK, Nelander K, Löfgren L, Nilsson RG and Cattaneo M: Ticagrelor inhibits human platelet aggregation via adenosine in addition to P2Y12 antagonism. J Thromb Haemost. 11:1867–1876. 2013.PubMed/NCBI
27	Heitzer T, Rudolph V, Schwedhelm E, Karstens M, Sydow K, Ortak M, Tschentscher P, Meinertz T, Böger R and Baldus S: Clopidogrel improves systemic endothelial nitric oxide bioavailability in patients with coronary artery disease: Evidence for antioxidant and antiinflammatory effects. Arterioscler Thromb Vasc Biol. 26:1648–1652. 2006. View Article : Google Scholar : PubMed/NCBI
28	Gwozdz P, Csanyi G, Luzak B, Gajda M, Mateuszuk L, Chmura-Skirlinska A, Watala C and Chlopicki S: Endothelial dysfunction and circulating platelet activation in apoE/LDLR/-mice along the development of atherosclerosis. In: Conference on Frontiers in Cardiovascular Biology. Cardiovasc Res. 87 Suppl:S94. 2010.
29	Wang W, Luo J, Xiang F, Liu X, Jiang M, Liao L and Hu J: Nucleolin down-regulation is involved in ADP-induced cell cycle arrest in S phase and cell apoptosis in vascular endothelial cells. PLoS One. 9:e1101012014. View Article : Google Scholar : PubMed/NCBI
30	Coutinho-Silva R, Stahl L, Cheung KK, de Campos NE, de Oliveira Souza C, Ojcius DM and Burnstock G: P2X and P2Y purinergic receptors on human intestinal epithelial carcinoma cells: Effects of extracellular nucleotides on apoptosis and cell proliferation. Am J Physiol Gastrointest Liver Physiol. 288:G1024–G1035. 2005. View Article : Google Scholar : PubMed/NCBI
31	Seidah NG, Benjannet S, Wickham L, Marcinkiewicz J, Jasmin SB, Stifani S, Basak A, Prat A and Chrétien M: The secretory proprotein convertase neural apoptosis-regulated convertase 1 (NARC-1): Liver regeneration and neuronal differentiation. Proc Natl Acad Sci USA. 100:928–933. 2003. View Article : Google Scholar : PubMed/NCBI
32	Kysenius K, Muggalla P, Mätlik K, Arumäe U and Huttunen HJ: PCSK9 regulates neuronal apoptosis by adjusting ApoER2 levels and signaling. Cell Mol Life Sci. 69:1903–1906. 2012. View Article : Google Scholar : PubMed/NCBI
33	Wu CY, Tang ZH, Jiang L, Li XF, Jiang ZS and Liu LS: PCSK9 siRNA inhibits HUVEC apoptosis induced by ox-LDL via Bcl/Bax-caspase9-caspase3 pathway. Mol Cell Biochem. 359:347–358. 2012. View Article : Google Scholar : PubMed/NCBI
34	Gurbel PA, Navarese EP and Tantry US: Exploration of PCSK9 as a cardiovascular risk factor: Is there a link to the platelet? J Am Coll Cardiol. 70:1463–1466. 2017. View Article : Google Scholar : PubMed/NCBI
35	Li S, Zhu CG, Guo YL, Xu RX, Zhang Y, Sun J and Li JJ: The relationship between the plasma PCSK9 levels and platelet indices in patients with stable coronary artery disease. J Atheroscler Thromb. 22:76–84. 2015. View Article : Google Scholar : PubMed/NCBI
36	Navarese EP, Kolodziejczak M, Winter MP, Alimohammadi A, Lang IM, Buffon A, Lip GY and Siller-Matula JM: Association of PCSK9 with platelet reactivity in patients with acute coronary syndrome treated with prasugrel or ticagrelor: The PCSK9-REACT study. Int J Cardiol. 227:644–649. 2017. View Article : Google Scholar : PubMed/NCBI
37	Chen TG, Chen TL, Chang HC, Tai YT, Cherng YG, Chang YT and Chen RM: Oxidized low-density lipoprotein induces apoptotic insults to mouse cerebral endothelial cells via a Bax-mitochondria-caspase protease pathway. Toxicol Appl Pharmacol. 219:42–53. 2007. View Article : Google Scholar : PubMed/NCBI
38	Yu W, Ying H, Tong F, Zhang C, Quan Y and Zhang Y: Protective effect of the silkworm protein 30Kc6 on human vascular endothelial cells damaged by oxidized low density lipoprotein (Ox-LDL). PLoS One. 8:e687462013. View Article : Google Scholar : PubMed/NCBI
39	Zhang L, Jia YH, Zhao XS, Zhou FH, Pan YY, Wan Q, Cui XB, Sun XG, Chen YY, Zhang Y and Cheng SB: Trichosanatine alleviates oxidized low-density lipoprotein induced endothelial cells injury via inhibiting the LOX-1/p38 MAPK pathway. Am J Transl Res. 8:5455–5464. 2016.PubMed/NCBI