Novel function of fluvastatin in attenuating oxidized low‑density lipoprotein‑induced endothelial cell ferroptosis in a glutathione peroxidase4‑ and cystine‑glutamate antiporter‑dependent manner

Li,Qing; Liu,Chang; Deng,Liang; Xie,Enrui; Yadav,Nishant; Tie,Yuanyuan; Cheng,Zheng; Deng,Jie

doi:10.3892/etm.2021.10710

Novel function of fluvastatin in attenuating oxidized low‑density lipoprotein‑induced endothelial cell ferroptosis in a glutathione peroxidase4‑ and cystine‑glutamate antiporter‑dependent manner

Authors:
- Qing Li
- Chang Liu
- Liang Deng
- Enrui Xie
- Nishant Yadav
- Yuanyuan Tie
- Zheng Cheng
- Jie Deng
View Affiliations

Affiliations: Department of Cardiology, The Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi 710004, P.R. China, The College of Traditional Chinese Medicine, Datong University, Datong, Shanxi 037009, P.R. China
Published online on: September 8, 2021 https://doi.org/10.3892/etm.2021.10710
Article Number: 1275
Copyright: © Li 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

Oxidized low‑density lipoprotein (ox‑LDL) induces endothelial cell apoptosis and dysfunction. Statins are drugs that are clinically used to lower serum cholesterol levels, and they have been shown to exert vascular protective effects. In the present study, human umbilical vein endothelial cells were transfected with scramble control siRNA or siRNA specific for glutathione peroxidase (GPx)4 or cystine‑glutamate antiporter (xCT). MTT, Matrigel and Transwell assays were used to evaluate cell proliferation, tube formation and migration, respectively. The levels of TNF‑α, IL‑α, 4‑hydroxynonenal, GPx4 and xCT expression were detected by western blot analysis. It was demonstrated that ox‑LDL promoted cytokine production and reduced the proliferation, migration and angiogenesis of endothelial cells. It was also observed that ox‑LDL decreased GPx4 and xCT expression and induced ferroptosis. Furthermore, the inhibition of ferroptosis by deferoxamine mesylate attenuated ox‑LDL‑induced endothelial cell dysfunction and restored ox‑LDL‑decreased GPx4 and xCT expression. Consistent with these results, GPx4 and xCT knockdown by siRNA transfection aggravated ox‑LDL‑induced endothelial cell dysfunction and inhibition of proliferation. To the best of our knowledge, the present study was the first to discover that fluvastatin may protect endothelial cells from ox‑LDL‑induced ferroptosis and dysfunction. Furthermore, knockdown of GPx4 and xCT expression blunted the protective effects of fluvastatin on ox‑LDL‑treated endothelial cells. These data indicated a novel function of fluvastatin in the protection of endothelial cells from ox‑LDL‑induced ferroptosis, the mechanism of which involves the regulation of GPx4 and xCT.

View Figures

View References

1	Khaddaj Mallat R, Mathew John CM, Kendrick DJ and Braun AP: The vascular endothelium: A regulator of arterial tone and interface for the immune system. Crit Rev Clin Lab Sci. 54:458–470. 2017.PubMed/NCBI View Article : Google Scholar
2	Rajendran P, Rengarajan T, Thangavel J, Nishigaki Y, Sakthisekaran D, Sethi G and Nishigaki I: The vascular endothelium and human diseases. Int J Biol Sci. 9:1057–1069. 2013.PubMed/NCBI View Article : Google Scholar
3	Beerepoot LV, Mehra N, Vermaat JS, Zonnenberg BA, Gebbink MF and Voest EE: Increased levels of viable circulating endothelial cells are an indicator of progressive disease in cancer patients. Ann Oncol. 15:139–145. 2004.PubMed/NCBI View Article : Google Scholar
4	Furstenberger G, von Moos R, Lucas R, Thürlimann B, Senn HJ, Hamacher J and Boneberg EM: Circulating endothelial cells and angiogenic serum factors during neoadjuvant chemotherapy of primary breast cancer. Br J Cancer. 94:524–531. 2006.PubMed/NCBI View Article : Google Scholar
5	Park KH and Park WJ: Endothelial dysfunction: Clinical implications in cardiovascular disease and therapeutic approaches. J Korean Med Sci. 30:1213–1225. 2015.PubMed/NCBI View Article : Google Scholar
6	Li TB, Zhang YZ, Liu WQ, Zhang JJ, Peng J, Luo XJ and Ma QL: Correlation between NADPH oxidase-mediated oxidative stress and dysfunction of endothelial progenitor cell in hyperlipidemic patients. Korean J Intern Med. 33:313–322. 2018.PubMed/NCBI View Article : Google Scholar
7	Valente AJ, Irimpen AM, Siebenlist U and Chandrasekar B: OxLDL induces endothelial dysfunction and death via TRAF3IP2: Inhibition by HDL3 and AMPK activators. Free Radic Biol Med. 70:117–128. 2014.PubMed/NCBI View Article : Google Scholar
8	Morawietz H: LOX-1 receptor as a novel target in endothelial dysfunction and atherosclerosis. Dtsch Med Wochenschr. 135:308–312. 2010.PubMed/NCBI View Article : Google Scholar : (In German).
9	Wahyudi S and Sargowo D: Green tea polyphenols inhibit oxidized LDL-induced NF-KB activation in human umbilical vein endothelial cells. Acta Med Indones. 39:66–70. 2007.PubMed/NCBI
10	Biocca S, Iacovelli F, Matarazzo S, Vindigni G, Oteri F, Desideri A and Falconi M: Molecular mechanism of statin-mediated LOX-1 inhibition. Cell Cycle. 14:1583–1595. 2015.PubMed/NCBI View Article : Google Scholar
11	Li J, Cao F, Yin HL, Huang ZJ, Lin ZT, Mao N, Sun B and Wang G: Ferroptosis: Past, present and future. Cell Death Dis. 11(88)2020.PubMed/NCBI View Article : Google Scholar
12	Maiorino M, Conrad M and Ursini F: GPx4, lipid peroxidation, and cell death: Discoveries, rediscoveries, and open issues. Antioxid Redox Signal. 29:61–74. 2018.PubMed/NCBI View Article : Google Scholar
13	Wortmann M, Schneider M, Pircher J, Hellfritsch J, Aichler M, Vegi N, Kölle P, Kuhlencordt P, Walch A, Pohl U, et al: Combined deficiency in glutathione peroxidase 4 and vitamin E causes multiorgan thrombus formation and early death in mice. Circ Res. 113:408–417. 2013.PubMed/NCBI View Article : Google Scholar
14	Ridker PM, Mora S, Rose L and Group JTS: Percent reduction in LDL cholesterol following high-intensity statin therapy: Potential implications for guidelines and for the prescription of emerging lipid-lowering agents. Eur Heart J. 37:1373–1379. 2016.PubMed/NCBI View Article : Google Scholar
15	Correction to: Pleiotropic effects of statins on the cardiovascular system. Circ Res. 123(e20)2018.
16	Wang J, Tokoro T, Matsui K, Higa S and Kitajima I: Pitavastatin at low dose activates endothelial nitric oxide synthase through PI3K-AKT pathway in endothelial cells. Life Sci. 76:2257–2268. 2005.PubMed/NCBI View Article : Google Scholar
17	Weis M, Heeschen C, Glassford AJ and Cooke JP: Statins have biphasic effects on angiogenesis. Circulation. 105:739–745. 2002.PubMed/NCBI View Article : Google Scholar
18	Stockwell BR, Friedmann Angeli JP, Bayir H, Bush AI, Conrad M, Dixon SJ, Fulda S, Gascón S, Hatzios SK, Kagan VE, et al: Ferroptosis: A regulated cell death nexus linking metabolism, redox biology, and disease. Cell. 171:273–285. 2017.PubMed/NCBI View Article : Google Scholar
19	Di Pietro N, Formoso G and Pandolfi A: Physiology and pathophysiology of oxLDL uptake by vascular wall cells in atherosclerosis. Vascul Pharmacol. 84:1–7. 2016.PubMed/NCBI View Article : Google Scholar
20	Yang WS, SriRamaratnam R, Welsch ME, Shimada K, Skouta R, Viswanathan VS, Cheah JH, Clemons PA, Shamji AF, Clish CB, et al: Regulation of ferroptotic cancer cell death by GPX4. Cell. 156:317–331. 2014.PubMed/NCBI View Article : Google Scholar
21	Wang Z, Ding Y, Wang X, Lu S, Wang C, He C, Wang L, Piao M, Chi G, Luo Y and Ge P: Pseudolaric acid B triggers ferroptosis in glioma cells via activation of Nox4 and inhibition of xCT. Cancer Lett. 428:21–33. 2018.PubMed/NCBI View Article : Google Scholar
22	Haruna Y, Morita Y, Yada T, Satoh M, Fox DA and Kashihara N: Fluvastatin reverses endothelial dysfunction and increased vascular oxidative stress in rat adjuvant-induced arthritis. Arthritis Rheum. 56:1827–1835. 2007.PubMed/NCBI View Article : Google Scholar
23	Ait-Oufella H, Maury E, Lehoux S, Guidet B and Offenstadt G: The endothelium: Physiological functions and role in microcirculatory failure during severe sepsis. Intensive Care Med. 36:1286–1298. 2010.PubMed/NCBI View Article : Google Scholar
24	Cosentino F, Rubattu S, Savoia C, Venturelli V, Pagannonne E and Volpe M: Endothelial dysfunction and stroke. J Cardiovasc Pharmacol. 38 (Suppl 2):S75–S78. 2001.PubMed/NCBI View Article : Google Scholar
25	Franses JW, Drosu NC, Gibson WJ, Chitalia VC and Edelman ER: Dysfunctional endothelial cells directly stimulate cancer inflammation and metastasis. Int J Cancer. 133:1334–1344. 2013.PubMed/NCBI View Article : Google Scholar
26	Widmer RJ and Lerman A: Endothelial dysfunction and cardiovascular disease. Glob Cardiol Sci Pract. 2014:291–308. 2014.PubMed/NCBI View Article : Google Scholar
27	Young MR: Endothelial cells in the eyes of an immunologist. Cancer Immunol Immunother. 61:1609–1616. 2012.PubMed/NCBI View Article : Google Scholar
28	Stroka KM, Levitan I and Aranda-Espinoza H: OxLDL and substrate stiffness promote neutrophil transmigration by enhanced endothelial cell contractility and ICAM-1. J Biomech. 45:1828–1834. 2012.PubMed/NCBI View Article : Google Scholar
29	Feng Y, Cai ZR, Tang Y, Hu G, Lu J, He D and Wang S: TLR4/NF-κB signaling pathway-mediated and oxLDL-induced up-regulation of LOX-1, MCP-1, and VCAM-1 expressions in human umbilical vein endothelial cells. Genet Mol Res. 13:680–695. 2014.PubMed/NCBI View Article : Google Scholar
30	Fang X, Wang H, Han D, Xie E, Yang X, Wei J, Gu S, Gao F, Zhu N, Yin X, et al: Ferroptosis as a target for protection against cardiomyopathy. Proc Natl Acad Sci USA. 116:2672–2680. 2019.PubMed/NCBI View Article : Google Scholar
31	Sui X, Zhang R, Liu S, Duan T, Zhai L, Zhang M, Han X, Xiang Y, Huang X, Lin H and Xie T: RSL3 drives ferroptosis through GPX4 inactivation and ROS production in colorectal cancer. Front Pharmacol. 9(1371)2018.PubMed/NCBI View Article : Google Scholar
32	Yu H, Guo P, Xie X, Wang Y and Chen G: Ferroptosis, a new form of cell death, and its relationships with tumourous diseases. J Cell Mol Med. 21:648–657. 2017.PubMed/NCBI View Article : Google Scholar
33	Adams SP, Sekhon SS, Tsang M and Wright JM: Fluvastatin for lowering lipids. Cochrane Database Syst Rev. 3(CD012282)2018.PubMed/NCBI View Article : Google Scholar
34	Lubrano V and Balzan S: Role of oxidative stress-related biomarkers in heart failure: Galectin 3, α1-antitrypsin and LOX-1: New therapeutic perspective? Mol Cell Biochem. 464:143–152. 2020.PubMed/NCBI View Article : Google Scholar
35	Singh S and Gautam AS: Upregulated LOX-1 receptor: Key player of the pathogenesis of atherosclerosis. Curr Atheroscler Rep. 21(38)2019.PubMed/NCBI View Article : Google Scholar
36	Zeya B and Chandra NC: LOX-1: Its cytotopographical variance and disease stress. J Biochem Mol Toxicol. 33(e22375)2019.PubMed/NCBI View Article : Google Scholar
37	Friedmann Angeli JP and Conrad M: Selenium and GPX4, a vital symbiosis. Free Radic Biol Med. 127:153–159. 2018.PubMed/NCBI View Article : Google Scholar
38	Shintoku R, Takigawa Y, Yamada K, Kubota C, Yoshimoto Y, Takeuchi T, Koshiishi I and Torii S: Lipoxygenase-mediated generation of lipid peroxides enhances ferroptosis induced by erastin and RSL3. Cancer Sci. 108:2187–2194. 2017.PubMed/NCBI View Article : Google Scholar