Autophagy in homocysteine‑induced HUVEC senescence

Zhang,Yexi; Ouyang,Juyan; Zhan,Liu; Li,Yu; Li,Shaoshan; He,Yi; Wang,Hong; Zhang,Xiangyang

doi:10.3892/etm.2023.12053

Autophagy in homocysteine‑induced HUVEC senescence

Authors:
- Yexi Zhang
- Juyan Ouyang
- Liu Zhan
- Yu Li
- Shaoshan Li
- Yi He
- Hong Wang
- Xiangyang Zhang
View Affiliations

Affiliations: Department of Functional Examination, Northern Jiangsu People's Hospital, The Affiliated Hospital of Yangzhou University, Yangzhou, Jiangsu 225000, P.R. China, Department of Geriatrics, The First Affiliated Hospital of Xinjiang Medical University, Urumchi, Xinjiang Uyghur Autonomous Region 830000, P.R. China, Department of Functional Science, Karamay College of Xinjiang Medical University, Karamay, Xinjiang Uyghur Autonomous Region 834000, P.R. China, Department of Clinical Teaching, Karamay College of Xinjiang Medical University, Karamay, Xinjiang Uyghur Autonomous Region 834000, P.R. China, Department of Morphology, Karamay College of Xinjiang Medical University, Karamay, Xinjiang Uyghur Autonomous Region 834000, P.R. China, Department of Cardiology, The First Affiliated Hospital of Xinjiang Medical University, Urumchi, Xinjiang Uyghur Autonomous Region 830000, P.R. China
Published online on: May 31, 2023 https://doi.org/10.3892/etm.2023.12053
Article Number: 354
Copyright: © Zhang 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

The senescence of vascular endothelial cells (VECs) drives the occurrence and development of cardiovascular disease (CVD). Homocysteine (HCY) is a general risk factor for age‑associated CVDs. Autophagy, an evolutionarily conserved lysosomal protein degradation pathway, serves a part in VEC senescence. The purpose of this study was to investigate the role of autophagy in HCY‑induced endothelial cell senescence and explore novel mechanisms and therapeutic approaches for related CVDs. Human umbilical vein endothelial cells (HUVECs) were isolated from fresh umbilical cords of healthy pregnancies. Cell Counting Kit‑8, flow cytometry and senescence‑associated (SA) β‑galactosidase (Gal) staining demonstrated that HCY induced HUVEC senescence by decreasing cell proliferation, arresting cell cycle and increasing the number of SA‑β‑Gal‑positive cells. Stub‑RFP‑Sens‑GFP‑LC3 autophagy‑related double fluorescence lentivirus revealed that HCY increased autophagic flux. Further, inhibition of autophagy using 3‑methyladenine increased HCY‑induced HUVEC senescence. By contrast, the induction of autophagy via rapamycin alleviated HCY‑induced HUVEC senescence. Finally, the detection of reactive oxygen species (ROS) with ROS kit showed that HCY increased intracellular ROS, whereas induction of autophagy reduced intracellular ROS. In conclusion, HCY increased HUVEC senescence and upregulated autophagy; moderate autophagy could reverse HCY‑induced cell senescence. Autophagy may alleviate HCY‑induced cell senescence by decreasing intracellular ROS. This provides insight into the underlying mechanism of HCY‑induced VEC senescence and potential treatments for age‑associated CVDs.

View Figures

View References

1	Donato AJ, Machin DR and Lesniewski LA: Mechanisms of dysfunction in the aging vasculature and role in age-related disease. Circ Res. 123:825–848. 2018.PubMed/NCBI View Article : Google Scholar
2	Ashapkin VV, Kutueva LI and Vanyushin BF: Aging as an epigenetic phenomenon. Curr Genomics. 18:385–407. 2017.PubMed/NCBI View Article : Google Scholar
3	Lakatta EG and Levy D: Arterial and cardiac aging: Major shareholders in cardiovascular disease enterprises: Part I: Aging arteries: A ‘set up’ for vascular disease. Circulation. 107:139–146. 2003.PubMed/NCBI View Article : Google Scholar
4	Sun C, Diao Q, Lu J, Zhang Z, Wu D, Wang X, Xie J, Zheng G, Shan Q, Fan S, et al: Purple sweet potato color attenuated NLRP3 inflammasome by inducing autophagy to delay endothelial senescence. J Cell Physiol. 234:5926–5939. 2019.PubMed/NCBI View Article : Google Scholar
5	Wang Y, Boerma M and Zhou D: Ionizing radiation-induced endothelial cell senescence and cardiovascular diseases. Radiat Res. 186:153–161. 2016.PubMed/NCBI View Article : Google Scholar
6	Moselhy SS and Demerdash SH: Plasma homocysteine and oxidative stress in cardiovascular disease. Dis Markers. 19:27–31. 2003.PubMed/NCBI View Article : Google Scholar
7	Scalera F, Martens-Lobenhoffer J, Täger M, Bukowska A, Lendeckel U and Bode-Böger SM: Effect of L-arginine on asymmetric dimethylarginine (ADMA) or homocysteine-accelerated endothelial cell aging. Biochem Biophys Res Commun. 345:1075–1082. 2006.PubMed/NCBI View Article : Google Scholar
8	Zhang D, Sun X, Liu J, Xie X, Cui W and Zhu Y: Homocysteine accelerates senescence of endothelial cells via DNA hypomethylation of human telomerase reverse transcriptase. Arterioscler Thromb Vasc Biol. 35:71–78. 2015.PubMed/NCBI View Article : Google Scholar
9	Wang CJ, Hu CP, Xu KP, Tan GS and Li YJ: Effects of selaginellin on homocysteine-induced senescence in human umbilical vein endothelial cells. J Cardiovasc Pharmacol. 55:560–566. 2010.PubMed/NCBI View Article : Google Scholar
10	Sun T, Ghosh AK, Eren M, Miyata T and Vaughan DE: PAI-1 contributes to homocysteine-induced cellular senescence. Cell Signal. 64(109394)2019.PubMed/NCBI View Article : Google Scholar
11	Abdellatif M, Sedej S, Carmona-Gutierrez D, Madeo F and Kroemer G: Autophagy in cardiovascular aging. Circ Res. 123:803–824. 2018.PubMed/NCBI View Article : Google Scholar
12	Boya P, Reggiori F and Codogno P: Emerging regulation and functions of autophagy. Nat Cell Biol. 15:713–720. 2013.PubMed/NCBI View Article : Google Scholar
13	Vion AC, Kheloufi M, Hammoutene A, Poisson J, Lasselin J, Devue C, Pic I, Dupont N, Busse J, Stark K, et al: Autophagy is required for endothelial cell alignment and atheroprotection under physiological blood flow. Proc Natl Acad Sci USA. 114:E8675–E8684. 2017.PubMed/NCBI View Article : Google Scholar
14	Song S, Wu S, Wang Y, Wang Z, Ye C, Song R, Song D and Ruan Y: 17β-estradiol inhibits human umbilical vascular endothelial cell senescence by regulating autophagy via p53. Exp Gerontol. 114:57–66. 2018.PubMed/NCBI View Article : Google Scholar
15	Zhang JW, Yan R, Tang YS, Guo YZ, Chang Y, Jing L, Wang YL and Zhang JZ: Hyperhomocysteinemia-induced autophagy and apoptosis with downregulation of hairy enhancer of split 1/5 in cortical neurons in mice. Int J Immunopathol Pharmacol. 30:371–382. 2017.PubMed/NCBI View Article : Google Scholar
16	Wang K, Peng S, Xiong S, Niu A, Xia M, Xiong X, Zeng G and Huang Q: Naringin inhibits autophagy mediated by PI3K-Akt-mTOR pathway to ameliorate endothelial cell dysfunction induced by high glucose/high fat stress. Eur J Pharmacol. 874(173003)2020.PubMed/NCBI View Article : Google Scholar
17	Jiang F: Autophagy in vascular endothelial cells. Clin Exp Pharmacol Physiol. 43:1021–1028. 2016.PubMed/NCBI View Article : Google Scholar
18	Guo ZX, Zhou FZ, Song W, Yu LL, Yan WJ, Yin LH, Sang H and Zhang HY: Suppression of microRNA-101 attenuates hypoxia-induced myocardial H9c2 cell injury by targeting DIMT1-Sp1/survivin pathway. Eur Rev Med Pharmacol Sci. 22:6965–6976. 2018.PubMed/NCBI View Article : Google Scholar
19	Khayati K, Antikainen H, Bonder EM, Weber GF, Kruger WD, Jakubowski H and Dobrowolski R: The amino acid metabolite homocysteine activates mTORC1 to inhibit autophagy and form abnormal proteins in human neurons and mice. FASEB J. 31:598–609. 2017.PubMed/NCBI View Article : Google Scholar
20	Nakano E, Taiwo FA, Nugent D, Griffiths HR, Aldred S, Paisi M, Kwok M, Bhatt P, Hill MH, Moat S and Powers HJ: Downstream effects on human low density lipoprotein of homocysteine exported from endothelial cells in an in vitro system. J Lipid Res. 46:484–493. 2005.PubMed/NCBI View Article : Google Scholar
21	Zhou C, Zhong W, Zhou J, Sheng F, Fang Z, Wei Y, Chen Y, Deng X, Xia B and Lin J: Monitoring autophagic flux by an improved tandem fluorescent-tagged LC3 (mTagRFP-mWasabi-LC3) reveals that high-dose rapamycin impairs autophagic flux in cancer cells. Autophagy. 8:1215–1226. 2012.PubMed/NCBI View Article : Google Scholar
22	Saxena S, Vekaria H, Sullivan PG and Seifert AW: Connective tissue fibroblasts from highly regenerative mammals are refractory to ROS-induced cellular senescence. Nat Commun. 10(4400)2019.PubMed/NCBI View Article : Google Scholar
23	Li L, Tan J, Miao Y, Lei P and Zhang Q: ROS and autophagy: Interactions and molecular regulatory mechanisms. Cell Mol Neurobiol. 35:615–621. 2015.PubMed/NCBI View Article : Google Scholar
24	Clarke R, Daly L, Robinson K, Naughten E, Cahalane S, Fowler B and Graham I: Hyperhomocysteinemia: An independent risk factor for vascular disease. N Engl J Med. 324:1149–1155. 1991.PubMed/NCBI View Article : Google Scholar
25	Esse R, Barroso M, Tavares de Almeida I and Castro R: The contribution of homocysteine metabolism disruption to endothelial dysfunction: State-of-the-art. Int J Mol Sci. 20(867)2019.PubMed/NCBI View Article : Google Scholar
26	Kwon Y, Kim JW, Jeoung JA, Kim MS and Kang C: Autophagy is pro-senescence when seen in close-up, but anti-senescence in long-shot. Mol Cells. 40:607–612. 2017.PubMed/NCBI View Article : Google Scholar
27	Tripathi M, Singh BK, Zhou J, Tikno K, Widjaja A, Sandireddy R, Arul K, Abdul Ghani SAB, Bee GGB, Wong KA, et al: Vitamin B(12) and folate decrease inflammation and fibrosis in NASH by preventing syntaxin 17 homocysteinylation. J Hepatol. 77:1246–1255. 2022.PubMed/NCBI View Article : Google Scholar
28	Wang M, Liang X, Cheng M, Yang L, Liu H, Wang X, Sai N and Zhang X: Homocysteine enhances neural stem cell autophagy in in vivo and in vitro model of ischemic stroke. Cell Death Dis. 10(561)2019.PubMed/NCBI View Article : Google Scholar
29	Zhang Y, Zhang Y, Tang J, Zhao S, Li C, Huang YP and Yi M: Oxymatrine inhibits homocysteine-mediated autophagy via MIF/mTOR signaling in human umbilical vein endothelial cells. Cell Physiol Biochem. 45:1893–1903. 2018.PubMed/NCBI View Article : Google Scholar