{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,6,29]],"date-time":"2026-06-29T20:58:02Z","timestamp":1782766682573,"version":"3.54.5"},"reference-count":52,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2017,5,5]],"date-time":"2017-05-05T00:00:00Z","timestamp":1493942400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81373002"],"award-info":[{"award-number":["81373002"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>The pathogenesis of atherosclerosis has been partly acknowledged to result from aberrant epigenetic mechanisms. Accordingly, low folate levels are considered to be a contributing factor to promoting vascular disease because of deregulation of DNA methylation. We hypothesized that increasing the levels of folic acid may act via an epigenetic gene silencing mechanism to ameliorate atherosclerosis. Here, we investigated the atheroprotective effects of folic acid and the resultant methylation status in high-fat diet-fed ApoE knockout mice and in oxidized low-density lipoprotein-treated human umbilical vein endothelial cells. We analyzed atherosclerotic lesion histology, folate concentration, homocysteine concentration, S-adenosylmethionine (SAM) and S-adenosylhomocysteine (SAH), and DNA methyltransferase activity, as well as monocyte chemotactic protein-1 (MCP1) and vascular endothelial growth factor (VEGF) expression and promoter methylation. Folic acid reduced atherosclerotic lesion size in ApoE knockout mice. The underlying folic acid protective mechanism appears to operate through regulating the normal homocysteine state, upregulating the SAM: SAH ratio, elevating DNA methyltransferase activity and expression, altering MCP1 and VEGF promoter methylation, and inhibiting MCP1 and VEGF expression. We conclude that folic acid supplementation effectively prevented atherosclerosis by modifying DNA methylation through the methionine cycle, improving DNA methyltransferase activity and expression, and thus changing the expression of atherosclerosis-related genes.<\/jats:p>","DOI":"10.3390\/ijms18050990","type":"journal-article","created":{"date-parts":[[2017,5,5]],"date-time":"2017-05-05T10:31:08Z","timestamp":1493980268000},"page":"990","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":47,"title":["Folic Acid Supplementation Delays Atherosclerotic Lesion Development by Modulating MCP1 and VEGF DNA Methylation Levels In Vivo and In Vitro"],"prefix":"10.3390","volume":"18","author":[{"given":"Shanshan","family":"Cui","sequence":"first","affiliation":[{"name":"Department of Nutrition and Food Science, School of Public Health, Tianjin Medical University, 22 Qixiangtai Road, Heping District, Tianjin 300070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wen","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Nutrition and Food Science, School of Public Health, Tianjin Medical University, 22 Qixiangtai Road, Heping District, Tianjin 300070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xin","family":"Lv","sequence":"additional","affiliation":[{"name":"Department of Nutrition and Food Science, School of Public Health, Tianjin Medical University, 22 Qixiangtai Road, Heping District, Tianjin 300070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pengyan","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Nutrition and Food Science, School of Public Health, Tianjin Medical University, 22 Qixiangtai Road, Heping District, Tianjin 300070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yuxia","family":"Gao","sequence":"additional","affiliation":[{"name":"Department of Cardiology, General Hospital of Tianjin Medical University, Tianjin 300052, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guowei","family":"Huang","sequence":"additional","affiliation":[{"name":"Department of Nutrition and Food Science, School of Public Health, Tianjin Medical University, 22 Qixiangtai Road, Heping District, Tianjin 300070, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2017,5,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"743","DOI":"10.1016\/S0140-6736(15)60692-4","article-title":"Global, regional, and national incidence, prevalence, and years lived with disability for 301 acute and chronic diseases and injuries in 188 countries, 1990\u20132013: A systematic analysis for the global burden of disease study 2013","volume":"386","author":"Vos","year":"2015","journal-title":"Lancet"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"868","DOI":"10.1038\/nature01323","article-title":"Inflammation in atherosclerosis","volume":"420","author":"Libby","year":"2002","journal-title":"Nature"},{"key":"ref_3","first-page":"1229","article-title":"Cytokine-activated human endothelial cells synthesize and secrete a monocyte chemoattractant, MCP-1\/JE","volume":"136","author":"Rollins","year":"1990","journal-title":"Am. J. Pathol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"e002851","DOI":"10.1161\/JAHA.115.002851","article-title":"Associations between macrophage colony-stimulating factor and monocyte chemotactic protein 1 in plasma and first-time coronary events: A nested case-control study","volume":"5","author":"Schiopu","year":"2016","journal-title":"J. Am. Heart Assoc."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"692","DOI":"10.1161\/CIRCGENETICS.113.000441","article-title":"DNA methylation map of human atherosclerosis","volume":"7","author":"Zaina","year":"2014","journal-title":"Circ. Cardiovasc. Genet."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"883","DOI":"10.1016\/j.mam.2012.08.001","article-title":"Cardiovascular epigenetics: From DNA methylation to micrornas","volume":"34","author":"Udali","year":"2013","journal-title":"Mol. Asp. Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"226","DOI":"10.1016\/j.tips.2015.02.005","article-title":"Epigenetic-related therapeutic challenges in cardiovascular disease","volume":"36","author":"Schiano","year":"2015","journal-title":"Trends Pharmacol. Sci."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1016\/S0092-8674(00)81656-6","article-title":"DNA methyltransferases DNMT3A and DNMT3B are essential for de novo methylation and mammalian development","volume":"99","author":"Okano","year":"1999","journal-title":"Cell"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"118","DOI":"10.1089\/biores.2012.0300","article-title":"Hyperhomocysteinemia-induced monocyte chemoattractant protein-1 promoter DNA methylation by nuclear factor-\u03baB\/DNA methyltransferase 1 in apolipoprotein E-deficient mice","volume":"2","author":"Wang","year":"2013","journal-title":"BioRes. Open Access"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"313","DOI":"10.4161\/epi.4.5.9160","article-title":"The expression of VEGF receptor genes is concurrently influenced by epigenetic gene silencing of the genes and VEGF activation","volume":"4","author":"Kim","year":"2009","journal-title":"Epigenetics"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"745","DOI":"10.1016\/j.ejim.2012.07.004","article-title":"Efficacy of folic acid supplementation in cardiovascular disease prevention: An updated meta-analysis of randomized controlled trials","volume":"23","author":"Yang","year":"2012","journal-title":"Eur. J. Intern. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1007\/s00394-014-0729-5","article-title":"Low-dose B vitamins supplementation ameliorates cardiovascular risk: A double-blind randomized controlled trial in healthy Chinese elderly","volume":"54","author":"Wang","year":"2015","journal-title":"Eur. J. Nutr."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"844","DOI":"10.1093\/molehr\/gav045","article-title":"Folate deficiency impairs decidualization and alters methylation patterns of the genome in mice","volume":"21","author":"Geng","year":"2015","journal-title":"Mol. Hum. Reprod."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"21","DOI":"10.3945\/an.111.000992","article-title":"Folate and DNA methylation: A review of molecular mechanisms and the evidence for folate\u2019s role","volume":"3","author":"Crider","year":"2012","journal-title":"Adv. Nutr."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"243","DOI":"10.1016\/j.ymgme.2014.10.006","article-title":"Homocysteine and DNA methylation: A review of animal and human literature","volume":"113","author":"Mandaviya","year":"2014","journal-title":"Mol. Genet. Metab."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"483","DOI":"10.1016\/j.numecd.2013.10.026","article-title":"B-vitamins intake, DNA-methylation of one carbon metabolism and homocysteine pathway genes and myocardial infarction risk: The epicor study","volume":"24","author":"Fiorito","year":"2014","journal-title":"Nutr. Metab. Cardiovasc. Dis."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"1335","DOI":"10.1161\/CIRCULATIONAHA.105.570846","article-title":"Improvement in stroke mortality in canada and the united states, 1990 to 2002","volume":"113","author":"Yang","year":"2006","journal-title":"Circulation"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"455","DOI":"10.1017\/S1368980008002565","article-title":"A cost-effectiveness analysis of folic acid fortification policy in the United States","volume":"12","author":"Bentley","year":"2009","journal-title":"Public Health Nutr."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"733","DOI":"10.12659\/MSM.889087","article-title":"Association between low-dose folic acid supplementation and blood lipids concentrations in male and female subjects with atherosclerosis risk factors","volume":"19","author":"Mierzecki","year":"2013","journal-title":"Med. Sci. Monit."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2674","DOI":"10.1161\/01.CIR.103.22.2674","article-title":"Kuopio Ischemic Heart Disease Risk Factor Study. Low dietary folate intake is associated with an excess incidence of acute coronary events: The kuopio ischemic heart disease risk factor study","volume":"103","author":"Voutilainen","year":"2001","journal-title":"Circulation"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1001\/jama.279.5.359","article-title":"Folate and vitamin B6 from diet and supplements in relation to risk of coronary heart disease among women","volume":"279","author":"Rimm","year":"1998","journal-title":"JAMA"},{"key":"ref_22","doi-asserted-by":"crossref","unstructured":"Zhou, Y.H., Tang, J.Y., Wu, M.J., Lu, J., Wei, X., Qin, Y.Y., Wang, C., Xu, J.F., and He, J. (2011). Effect of folic acid supplementation on cardiovascular outcomes: A systematic review and meta-analysis. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0025142"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1205","DOI":"10.1161\/STROKEAHA.109.573410","article-title":"Efficacy of homocysteine-lowering therapy with folic acid in stroke prevention: A meta-analysis","volume":"41","author":"Lee","year":"2010","journal-title":"Stroke"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"4580","DOI":"10.2174\/1381612820666140522115801","article-title":"Impact of MCP-1 in atherosclerosis","volume":"20","author":"Lin","year":"2014","journal-title":"Curr. Pharm. Des."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"894","DOI":"10.1038\/29788","article-title":"Decreased lesion formation in CCR2\u2212\/\u2212 mice reveals a role for chemokines in the initiation of atherosclerosis","volume":"394","author":"Boring","year":"1998","journal-title":"Nature"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"817","DOI":"10.1007\/s00011-011-0337-2","article-title":"Folic acid inhibits lipopolysaccharide-induced inflammatory response in raw264.7 macrophages by suppressing MAPKs and NK-\u03baB activation","volume":"60","author":"Feng","year":"2011","journal-title":"Inflamm. Res."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"627","DOI":"10.1007\/s10620-008-0398-4","article-title":"Increased folate intake with no changes in serum homocysteine and decreased levels of C-reactive protein in patients with inflammatory bowel diseases","volume":"54","author":"Chiarello","year":"2009","journal-title":"Dig. Dis. Sci."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"269","DOI":"10.5543\/tkda.2011.01290","article-title":"The relation of serum monocyte chemoattractant protein-1 level with coronary atherosclerotic burden and collateral degree in stable coronary artery disease","volume":"39","author":"Sahinarslan","year":"2011","journal-title":"Turk Kardiyol. Dern. Arsivi"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"29044","DOI":"10.1074\/jbc.M114.555235","article-title":"Genome-wide approaches reveal functional vascular endothelial growth factor (VEGF)-inducible nuclear factor of activated T cells (NFAT) c1 binding to angiogenesis-related genes in the endothelium","volume":"289","author":"Suehiro","year":"2014","journal-title":"J. Biol. Chem."},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"Liu, X.L., Zhang, P.F., Ding, S.F., Wang, Y., Zhang, M., Zhao, Y.X., Ni, M., and Zhang, Y. (2012). Local gene silencing of monocyte chemoattractant protein-1 prevents vulnerable plaque disruption in apolipoprotein E-knockout mice. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0033497"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1517\/14712598.2012.641011","article-title":"Pro- and anti-angiogenic therapy and atherosclerosis with special emphasis on vascular endothelial growth factors","volume":"12","author":"Vuorio","year":"2012","journal-title":"Expert Opin. Biol. Ther."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1002\/biof.1306","article-title":"Modulation of the vascular endothelium functioning by dietary components, the role of epigenetics","volume":"43","author":"Wojtala","year":"2017","journal-title":"Biofactors"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"14","DOI":"10.1186\/s13148-015-0046-8","article-title":"Vitamin B12 insufficiency induces cholesterol biosynthesis by limiting S-adenosylmethionine and modulating the methylation of SREBF1 and LDLR genes","volume":"7","author":"Adaikalakoteswari","year":"2015","journal-title":"Clin. Epigenetics"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/j.cca.2016.02.026","article-title":"Aberrant DNA methylation in the pathogenesis of atherosclerosis","volume":"456","author":"Hai","year":"2016","journal-title":"Clin. Chim. Acta"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"172","DOI":"10.1002\/mnfr.201200636","article-title":"Methyl nutrients, DNA methylation, and cardiovascular disease","volume":"58","author":"Glier","year":"2014","journal-title":"Mol. Nutr. Food Res."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"2811","DOI":"10.1093\/jn\/131.11.2811","article-title":"Intracellular S-adenosylhomocysteine concentrations predict global DNA hypomethylation in tissues of methyl-deficient cystathionine \u03b2-synthase heterozygous mice","volume":"131","author":"Caudill","year":"2001","journal-title":"J. Nutr."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"158","DOI":"10.1016\/j.biocel.2015.06.015","article-title":"Role of S-adenosylhomocysteine in cardiovascular disease and its potential epigenetic mechanism","volume":"67","author":"Xiao","year":"2015","journal-title":"Int. J. Biochem. Cell Biol."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"92","DOI":"10.2174\/092986708783330700","article-title":"Mammalian cytosine DNA methyltransferase DNMT1: Enzymatic mechanism, novel mechanism-based inhibitors, and RNA-directed DNA methylation","volume":"15","author":"Svedruzic","year":"2008","journal-title":"Curr. Med. Chem."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"576","DOI":"10.1002\/mrd.21344","article-title":"Improved in vitro development of cloned bovine embryos using S-adenosylhomocysteine, a non-toxic epigenetic modifying reagent","volume":"78","author":"Jafari","year":"2011","journal-title":"Mol. Reprod. Dev."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"883","DOI":"10.1016\/j.jnutbio.2015.03.009","article-title":"Folic acid administration inhibits amyloid \u03b2-peptide accumulation in APP\/PS1 transgenic mice","volume":"26","author":"Li","year":"2015","journal-title":"J. Nutr. Biochem."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"730","DOI":"10.1161\/STROKEAHA.108.526798","article-title":"High-dose B vitamin supplementation and progression of subclinical atherosclerosis: A randomized controlled trial","volume":"40","author":"Hodis","year":"2009","journal-title":"Stroke"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"2142","DOI":"10.1093\/eurheartj\/ehm272","article-title":"Contribution of changes in incidence and mortality to trends in the prevalence of coronary heart disease in the UK: 1996\u20132005","volume":"28","author":"Davies","year":"2007","journal-title":"Eur. Heart J."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1019","DOI":"10.1161\/01.HYP.0000223064.62762.0b","article-title":"Gender differences of cardiovascular disease: New perspectives for estrogen receptor signaling","volume":"47","author":"Meyer","year":"2006","journal-title":"Hypertension"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"136","DOI":"10.1007\/s00392-006-0351-5","article-title":"Gender differences in the metabolic syndrome and their role for cardiovascular disease","volume":"95","author":"Lehmkuhl","year":"2006","journal-title":"Clin. Res. Cardiol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/0091-7435(91)90006-P","article-title":"Estrogen replacement therapy and coronary heart disease: A quantitative assessment of the epidemiologic evidence","volume":"20","author":"Stampfer","year":"1991","journal-title":"Prev. Med."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"301","DOI":"10.1016\/S0021-9150(99)00081-7","article-title":"Effects of sex and age on atherosclerosis and autoimmunity in ApoE-deficient mice","volume":"145","author":"Caligiuri","year":"1999","journal-title":"Atherosclerosis"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"16457","DOI":"10.1073\/pnas.1104533108","article-title":"Myeloid-specific estrogen receptor \u03b1 deficiency impairs metabolic homeostasis and accelerates atherosclerotic lesion development","volume":"108","author":"Ribas","year":"2011","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_48","first-page":"5","article-title":"Animal models for the study of the relationships between diet and obesity: A focus on dietary protein and estrogen deficiency","volume":"4","author":"Blachier","year":"2017","journal-title":"Front. Nutr."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Han, J.M., Li, H., Cho, M.H., Baek, S.H., Lee, C.H., Park, H.Y., and Jeong, T.S. (2017). Soy-leaf extract exerts atheroprotective effects via modulation of kruppel-like factor 2 and adhesion molecules. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18020373"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"76423","DOI":"10.18632\/oncotarget.13121","article-title":"Tet methylcytosine dioxygenase 2 inhibits atherosclerosis via upregulation of autophagy in ApoE\u2212\/\u2212 mice","volume":"7","author":"Peng","year":"2016","journal-title":"Oncotarget"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1693","DOI":"10.1016\/j.ajpath.2011.06.037","article-title":"Paigen diet-fed apolipoprotein E knockout mice develop severe pulmonary hypertension in an interleukin-1-dependent manner","volume":"179","author":"Lawrie","year":"2011","journal-title":"Am. J. Pathol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1007\/978-1-4939-2929-0_5","article-title":"Oil red O and hematoxylin and eosin staining for quantification of atherosclerosis burden in mouse aorta and aortic root","volume":"1339","author":"Andres","year":"2015","journal-title":"Methods Mol. Biol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/18\/5\/990\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T18:34:46Z","timestamp":1760207686000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/18\/5\/990"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,5,5]]},"references-count":52,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2017,5]]}},"alternative-id":["ijms18050990"],"URL":"https:\/\/doi.org\/10.3390\/ijms18050990","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,5,5]]}}}