Homocysteine injures vascular endothelial cells by inhibiting mitochondrial activity

Yang,Fengyong; Qi,Xiujing; Gao,Zheng; Yang,Xingju; Zheng,Xingfeng; Duan,Chonghao; Zheng,Jian

doi:10.3892/etm.2016.3564

Homocysteine injures vascular endothelial cells by inhibiting mitochondrial activity

Authors:
- Fengyong Yang
- Xiujing Qi
- Zheng Gao
- Xingju Yang
- Xingfeng Zheng
- Chonghao Duan
- Jian Zheng
View Affiliations

Affiliations: Intensive Care Unit, The People's Hospital of Laiwu, Laiwu, Shandong 271199, P.R. China, Burn Center, Shanghai Hospital, Second Military Medical University, Shanghai 200433, P.R. China, Department of Thoracic Surgery, The People's Hospital of Laicheng, Laiwu, Shandong 271199, P.R. China
Published online on: August 2, 2016 https://doi.org/10.3892/etm.2016.3564
Pages: 2247-2252

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

The aim of the present study was to investigate the role of homocysteine (Hcy) in the pathogenesis of pulmonary embolism (PE) and the associated molecular mechanisms in human umbilical vein endothelial cells (HUVECs). Hcy contents were detected with high‑performance liquid chromatography. Apoptosis was detected by flow cytometry using Annexin‑V staining. Cytochrome c oxidase (COX) activity was assessed with an enzyme activity assay, and the expression levels of COX 17 were determined by western blot analysis. Intracellular reactive oxygen species levels were measured using a microplate reader with a fluorescence probe. The results demonstrated that, compared with the control group, the serum Hcy levels were significantly elevated in the PE group, suggesting that Hcy may be an indicator for PE. Following treatment with Hcy, the apoptosis rate was markedly elevated in HUVECs. Moreover, Hcy decreased COX activity and downregulated the expression of COX 17 in HUVECs. Furthermore, Hcy increased the ROS levels in these endothelial cells. However, all the above‑mentioned physiopathological changes induced by Hcy in HUVECs could be restored by folic acid. In conclusion, the results of the present study demonstrated that Hcy inhibited COX activity, downregulated COX 17 expression, increased intracellular ROS levels and enhanced apoptosis in endothelial cells.

View Figures

View References

1	de Miguel-Díez J, Jiménez-García R, de Andrés A López, Hernández-Barrera V, Carrasco-Garrido P, Monreal M, Jiménez D, Jara-Palomares L and Álvaro-Meca A: Analysis of environmental risk factors for pulmonary embolism: A case-crossover study (2001-2013). Eur J Intern Med. 31:55–61. 2016. View Article : Google Scholar : PubMed/NCBI
2	Heit JA: The epidemiology of venous thromboembolism in the community: Implications for prevention and management. J Thromb Thrombolysis. 21:23–29. 2006. View Article : Google Scholar : PubMed/NCBI
3	Tapson VF: Acute pulmonary embolism. N Engl J Med. 358:1037–1052. 2008. View Article : Google Scholar : PubMed/NCBI
4	Chen P, Poddar R, Tipa EV, Dibello PM, Moravec CD, Robinson K, Green R, Kruger WD, Garrow TA and Jacobsen DW: Homocysteine metabolism in cardiovascular cells and tissues: Implications for hyperhomocysteinemia and cardiovascular disease. Adv Enzyme Regul. 39:93–109. 1999. View Article : Google Scholar : PubMed/NCBI
5	Jeremy JY, Shukla N, Angelini GD, Day A, Wan IY, Talpahewa SP and Ascione R: Sustained increases of plasma homocysteine, copper and serum ceruloplasmin after coronary artery bypass grafting. Ann Thorac Surg. 74:1553–1557. 2002. View Article : Google Scholar : PubMed/NCBI
6	Karalezli A, Parlak ES, Kanbay A, Senturk A and Hasanoglu HC: Homocysteine and serum-lipid levels in pulmonary embolism. Clin Appl Thromb Hemost. 17:E186–E189. 2011. View Article : Google Scholar : PubMed/NCBI
7	Köktürk N, Kanbay A, Aydoğdu M, Özyılmaz E, Bukan N and Ekim N: Hyperhomocysteinemia prevalence among patients with venous thromboembolism. Clin Appl Thromb Hemost. 17:487–493. 2011. View Article : Google Scholar : PubMed/NCBI
8	Okumus G, Kiyan E, Arseven O, Tabak L, DizKucukkaya R, Unlucerci Y, Abaci N, Unaltuna NE and Issever H: Hereditary thrombophilic risk factors and venous thromboembolism in Istanbul, Turkey: The role in different clinical manifestations of venous thromboembolism. Clin Appl Thromb Hemost. 14:168–173. 2008. View Article : Google Scholar : PubMed/NCBI
9	SallesCrawley II, Monkman JH, Ahnström J, Lane DA and Crawley JT: Vessel wall BAMBI contributes to hemostasis and thrombus stability. Blood. 123:2873–2881. 2014. View Article : Google Scholar : PubMed/NCBI
10	Ren M, Li R, Luo M, Chen N, Deng X, Yan K, Zeng M and Wu J: Endothelial cells but not platelets are the major source of Toll-like receptor 4 in the arterial thrombosis and tissue factor expression in mice. Am J Physiol Regul Integr Comp Physiol. 307:R901–R907. 2014. View Article : Google Scholar : PubMed/NCBI
11	Mansoor MA, Bergmark C, Haswell SJ, Savage IF, Evans PH, Berge RK, Svardal AM and Kristensen O: Correlation between plasma total homocysteine and copper in patients with peripheral vascular disease. Clin Chem. 46:385–391. 2000.PubMed/NCBI
12	Shukla N, Angelini GD and Jeremy JY: Interactive effects of homocysteine and copper on angiogenesis in porcine isolated saphenous vein. Ann Thorac Surg. 84:43–49. 2007. View Article : Google Scholar : PubMed/NCBI
13	Garnier A, Fortin D, Deloménie C, Momken I, Veksler V and Ventura-Clapier R: Depressed mitochondrial transcription factors and oxidative capacity in rat failing cardiac and skeletal muscles. J Physiol. 551:491–501. 2003. View Article : Google Scholar : PubMed/NCBI
14	Cawthon D, McNew R, Beers KW and Bottje WG: Evidence of mitochondrial dysfunction in broilers with pulmonary hypertension syndrome (Ascites): Effect of t-butyl hydroperoxide on hepatic mitochondrial function, glutathione and related thiols. Poult Sci. 78:114–124. 1999. View Article : Google Scholar : PubMed/NCBI
15	Cawthon D, Beers K and Bottje WG: Electron transport chain defect and inefficient respiration may underlie pulmonary hypertension syndrome (ascites)-associated mitochondrial dysfunction in broilers. Poult Sci. 80:474–484. 2001. View Article : Google Scholar : PubMed/NCBI
16	Suter M, Remé C, Grimm C, Wenzel A, Jäättela M, Esser P, Kociok N, Leist M and Richter C: Age-related macular degeneration. The lipofusion component N-retinyl-N-retinylidene ethanolamine detaches proapoptotic proteins from mitochondria and induces apoptosis in mammalian retinal pigment epithelial cells. J Biol Chem. 275:39625–39630. 2000. View Article : Google Scholar : PubMed/NCBI
17	Srinivasan S and Avadhani NG: Cytochrome c oxidase dysfunction in oxidative stress. Free Radic Biol Med. 53:1252–1263. 2012. View Article : Google Scholar : PubMed/NCBI
18	Leadsham JE, Sanders G, Giannaki S, Bastow EL, Hutton R, Naeimi WR, Breitenbach M and Gourlay CW: Loss of cytochrome c oxidase promotes RAS-dependent ROS production from the ER resident NADPH oxidase, Yno1p, in yeast. Cell Metab. 18:279–286. 2013. View Article : Google Scholar : PubMed/NCBI
19	Zou H, Li Y, Liu X and Wang X: An APAF-1. cytochrome c multimeric complex is a functional apoptosome that activates procaspase-9. J Biol Chem. 274:11549–11556. 1999. View Article : Google Scholar : PubMed/NCBI
20	Hu Y, Benedict MA, Ding L and Núñez G: Role of cytochrome c and dATP/ATP hydrolysis in Apaf-1-mediated caspase-9 activation and apoptosis. EMBO J. 18:3586–3595. 1999. View Article : Google Scholar : PubMed/NCBI
21	Saleh A, Srinivasula SM, Acharya S, Fishel R and Alnemri ES: Cytochrome c and dATP-mediated oligomerization of Apaf-1 is a prerequisite for procaspase-9 activation. J Biol Chem. 274:17941–17945. 1999. View Article : Google Scholar : PubMed/NCBI
22	Hassanin IM, Shahin AY, Badawy MS and Karam K: D-dimer testing versus multislice computed tomography in the diagnosis of postpartum pulmonary embolism in symptomatic high-risk women. Int J Gynaecol Obstet. 115:200–201. 2011. View Article : Google Scholar : PubMed/NCBI
23	Glerum DM, Shtanko A and Tzagoloff A: Characterization of COX17, a yeast gene involved in copper metabolism and assembly of cytochrome oxidase. J Biol Chem. 271:14504–14509. 1996. View Article : Google Scholar : PubMed/NCBI
24	Takahashi Y, Kako K, Kashiwabara S, Takehara A, Inada Y, Arai H, Nakada K, Kodama H, Hayashi J, Baba T and Munekata E: Mammalian copper chaperone Cox17p has an essential role in activation of cytochrome C oxidase and embryonic development. Mol Cell Biol. 22:7614–7621. 2002. View Article : Google Scholar : PubMed/NCBI