{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,15]],"date-time":"2026-05-15T13:51:28Z","timestamp":1778853088867,"version":"3.51.4"},"reference-count":57,"publisher":"Public Library of Science (PLoS)","issue":"5","license":[{"start":{"date-parts":[[2025,5,29]],"date-time":"2025-05-29T00:00:00Z","timestamp":1748476800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS One"],"abstract":"<jats:p>Decreased nitric oxide (NO) production from the vascular endothelium is a major factor for vascular aging. Because vascular aging has few specific subjective symptoms, assessing the susceptibility to vascular aging is beneficial for its early detection and improvement. Therefore, this study evaluated the associations between single nucleotide polymorphisms (SNPs) on L-citrulline (Cit) pathways essential for NO production and the health characteristics involved in vascular aging using a candidate gene approach. Associations with a significance level included those between the <jats:italic>KCNMB4<\/jats:italic> rs17108108 C allele and tendency to gain weight, the <jats:italic>ADCY8<\/jats:italic> rs6470860 G allele and numbness of limbs, the <jats:italic>NOS1<\/jats:italic> rs2271987 T allele and lower back pain, and the <jats:italic>PDE9A<\/jats:italic> rs2284972 G allele and body pain with negative mood states. A genome-wide association study was also conducted to analyze SNPs more extensively across genes related to Cit and NO metabolism, which revealed genome-wide significant associations between the <jats:italic>PRMT6<\/jats:italic> rs12028323 C allele and mood disturbance. These significant associations could be explained by the change in downstream NO signaling, supporting the relationship between the investigated traits and vascular function. These traits can be manifested as subjective symptoms of vascular aging. Therefore, the identified SNPs could predict susceptibility to the subjective symptoms of vascular aging, which could lead to genotype-based personalized interventions of Cit for an efficient improvement of vascular health.<\/jats:p>","DOI":"10.1371\/journal.pone.0323778","type":"journal-article","created":{"date-parts":[[2025,5,29]],"date-time":"2025-05-29T17:25:01Z","timestamp":1748539501000},"page":"e0323778","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":3,"title":["SNP associations in the L-citrulline metabolic pathway and vascular aging in the Japanese population"],"prefix":"10.1371","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0009-0008-8477-288X","authenticated-orcid":true,"given":"Dai","family":"Nogimura","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kazuki","family":"Moriyasu","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6463-3059","authenticated-orcid":true,"given":"Sachiko","family":"Ishida","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Masakazu","family":"Kohda","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takayuki","family":"Yazawa","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0009-0007-9394-6135","authenticated-orcid":true,"given":"Masahiko","family":"Morita","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2025,5,29]]},"reference":[{"issue":"4","key":"pone.0323778.ref001","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1007\/s00109-015-1265-3","article-title":"Role of sGC-dependent NO signalling and myocardial infarction risk","volume":"93","author":"J Wobst","year":"2015","journal-title":"J Mol Med (Berl)"},{"issue":"8","key":"pone.0323778.ref002","doi-asserted-by":"crossref","first-page":"1949","DOI":"10.1016\/j.bbadis.2018.11.007","article-title":"Functions and dysfunctions of nitric oxide in brain","volume":"1865","author":"P Pic\u00f3n-Pag\u00e8s","year":"2019","journal-title":"Biochim Biophys Acta Mol Basis Dis"},{"issue":"5","key":"pone.0323778.ref003","doi-asserted-by":"crossref","first-page":"1652","DOI":"10.1152\/japplphysiol.90549.2008","article-title":"Effects of central arterial aging on the structure and function of the peripheral vasculature: implications for end-organ damage","volume":"105","author":"GF Mitchell","year":"2008","journal-title":"J Appl Physiol (1985)"},{"issue":"1","key":"pone.0323778.ref004","doi-asserted-by":"crossref","first-page":"113","DOI":"10.1053\/j.gastro.2005.04.014","article-title":"The natural history of nonalcoholic fatty liver disease: a population-based cohort study","volume":"129","author":"LA Adams","year":"2005","journal-title":"Gastroenterology"},{"issue":"20","key":"pone.0323778.ref005","doi-asserted-by":"crossref","first-page":"10666","DOI":"10.3390\/ijerph182010666","article-title":"Vascular ageing and aerobic exercise","volume":"18","author":"M Kozakova","year":"2021","journal-title":"Int J Environ Res Public Health"},{"issue":"2","key":"pone.0323778.ref006","first-page":"20","article-title":"cGMP signaling and vascular smooth muscle cell plasticity","volume":"5","author":"M Lehners","year":"2018","journal-title":"J Cardiovasc Dev Dis"},{"issue":"5","key":"pone.0323778.ref007","doi-asserted-by":"crossref","first-page":"1268","DOI":"10.3390\/nu15051268","article-title":"Dietary arginine and citrulline supplements for cardiovascular health and athletic performance: a narrative review","volume":"15","author":"H-Y Park","year":"2023","journal-title":"Nutrients"},{"issue":"3","key":"pone.0323778.ref008","doi-asserted-by":"crossref","first-page":"214","DOI":"10.2174\/18715222113139990008","article-title":"Effects of oral L-citrulline supplementation on lipoprotein oxidation and endothelial dysfunction in humans with vasospastic angina","volume":"13","author":"M Morita","year":"2013","journal-title":"Immunol Endocr Metab Agents Med Chem"},{"issue":"2","key":"pone.0323778.ref009","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1016\/j.ijcard.2010.10.004","article-title":"Short-term effects of L-citrulline supplementation on arterial stiffness in middle-aged men","volume":"155","author":"M Ochiai","year":"2012","journal-title":"Int J Cardiol"},{"issue":"1","key":"pone.0323778.ref010","doi-asserted-by":"crossref","first-page":"352","DOI":"10.1093\/nar\/28.1.352","article-title":"dbSNP: a database of single nucleotide polymorphisms","volume":"28","author":"EM Smigielski","year":"2000","journal-title":"Nucleic Acids Res"},{"issue":"3","key":"pone.0323778.ref011","doi-asserted-by":"crossref","first-page":"297","DOI":"10.1101\/pdb.top068163","article-title":"Introduction to genetic association studies","volume":"2012","author":"CM Lewis","year":"2012","journal-title":"Cold Spring Harb Protoc"},{"issue":"2","key":"pone.0323778.ref012","doi-asserted-by":"crossref","first-page":"195","DOI":"10.1016\/j.ajhg.2022.12.017","article-title":"Using genetic association data to guide drug discovery and development: review of methods and applications","volume":"110","author":"S Burgess","year":"2023","journal-title":"Am J Hum Genet"},{"key":"pone.0323778.ref013","article-title":"Personalized nutrition: a review of genotype-based nutritional supplementation","volume":"9","author":"F Wang","year":"2022","journal-title":"Front Nutr"},{"issue":"9791","key":"pone.0323778.ref014","doi-asserted-by":"crossref","first-page":"584","DOI":"10.1016\/S0140-6736(11)60872-6","article-title":"Effect modification by population dietary folate on the association between MTHFR genotype, homocysteine, and stroke risk: a meta-analysis of genetic studies and randomised trials","volume":"378","author":"MV Holmes","year":"2011","journal-title":"Lancet"},{"key":"pone.0323778.ref015","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1146\/annurev-nutr-082018-124250","article-title":"FADS1 and FADS2 polymorphisms modulate fatty acid metabolism and dietary impact on health","volume":"39","author":"B Koletzko","year":"2019","journal-title":"Annu Rev Nutr"},{"issue":"10","key":"pone.0323778.ref016","doi-asserted-by":"crossref","first-page":"1162","DOI":"10.1038\/ng.3660","article-title":"Meta-analysis identifies common and rare variants influencing blood pressure and overlapping with metabolic trait loci","volume":"48","author":"C Liu","year":"2016","journal-title":"Nat Genet"},{"issue":"2","key":"pone.0323778.ref017","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1161\/HYPERTENSIONAHA.119.13513","article-title":"Influence of genetic variation in PDE3A on endothelial function and stroke","volume":"75","author":"M Traylor","year":"2020","journal-title":"Hypertension"},{"issue":"7","key":"pone.0323778.ref018","doi-asserted-by":"crossref","first-page":"420","DOI":"10.7150\/ijbs.3.420","article-title":"Candidate gene identification approach: progress and challenges","volume":"3","author":"M Zhu","year":"2007","journal-title":"Int J Biol Sci"},{"issue":"1","key":"pone.0323778.ref019","doi-asserted-by":"crossref","DOI":"10.1038\/s43586-021-00056-9","article-title":"Genome-wide association studies","volume":"1","author":"E Uffelmann","year":"2021","journal-title":"Nat Rev Methods Primers"},{"issue":"3","key":"pone.0323778.ref020","doi-asserted-by":"crossref","first-page":"559","DOI":"10.1086\/519795","article-title":"PLINK: a tool set for whole-genome association and population-based linkage analyses","volume":"81","author":"S Purcell","year":"2007","journal-title":"Am J Hum Genet"},{"issue":"11","key":"pone.0323778.ref021","doi-asserted-by":"crossref","first-page":"1443","DOI":"10.1038\/ng.3679","article-title":"Reference-based phasing using the haplotype reference consortium panel","volume":"48","author":"P-R Loh","year":"2016","journal-title":"Nat Genet"},{"issue":"10","key":"pone.0323778.ref022","doi-asserted-by":"crossref","first-page":"1284","DOI":"10.1038\/ng.3656","article-title":"Next-generation genotype imputation service and methods","volume":"48","author":"S Das","year":"2016","journal-title":"Nat Genet"},{"issue":"7571","key":"pone.0323778.ref023","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1038\/nature15393","article-title":"A global reference for human genetic variation","volume":"526","author":"1000 Genomes Project Consortium","year":"2015","journal-title":"Nature"},{"issue":"6","key":"pone.0323778.ref024","doi-asserted-by":"crossref","first-page":"603","DOI":"10.1080\/10615806.2011.582948","article-title":"Transient state-dependent fluctuations in anxiety measured using STAI, POMS, PANAS or VAS: a comparative review","volume":"25","author":"V Rossi","year":"2012","journal-title":"Anxiety Stress Coping"},{"issue":"11","key":"pone.0323778.ref025","doi-asserted-by":"crossref","first-page":"1037","DOI":"10.1016\/S0895-4356(98)00095-X","article-title":"Translation, adaptation, and validation of the SF-36 health survey for use in Japan","volume":"51","author":"S Fukuhara","year":"1998","journal-title":"J Clin Epidemiol"},{"issue":"2","key":"pone.0323778.ref026","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1097\/PSY.0b013e3181cdbfc0","article-title":"Adverse impact of mood on flow-mediated dilation","volume":"72","author":"DC Cooper","year":"2010","journal-title":"Psychosom Med"},{"issue":"3","key":"pone.0323778.ref027","doi-asserted-by":"crossref","first-page":"254","DOI":"10.1016\/j.amjms.2018.06.014","article-title":"Efficacy and safety of policosanol plus fenofibrate combination therapy in elderly patients with mixed dyslipidemia: a randomized, controlled clinical study","volume":"356","author":"H-Y Wang","year":"2018","journal-title":"Am J Med Sci"},{"issue":"7889","key":"pone.0323778.ref028","doi-asserted-by":"crossref","first-page":"1127","DOI":"10.1016\/S0140-6736(74)90884-8","article-title":"Measurement of pain","volume":"2","author":"EC Huskisson","year":"1974","journal-title":"Lancet Lond Engl"},{"issue":"9","key":"pone.0323778.ref029","doi-asserted-by":"crossref","first-page":"2411","DOI":"10.1007\/s00421-013-2678-8","article-title":"The contribution of blood flow to the skin temperature responses during a cold sensitivity test","volume":"113","author":"M Davey","year":"2013","journal-title":"Eur J Appl Physiol"},{"key":"pone.0323778.ref030","doi-asserted-by":"crossref","first-page":"642710","DOI":"10.3389\/fmed.2021.642710","article-title":"Reduced endothelial function in myalgic encephalomyelitis\/chronic fatigue syndrome-results from open-label cyclophosphamide intervention study","volume":"8","author":"K S\u00f8rland","year":"2021","journal-title":"Front Med (Lausanne)"},{"issue":"3","key":"pone.0323778.ref031","doi-asserted-by":"crossref","first-page":"267","DOI":"10.5935\/1984-0063.20220050","article-title":"Association of sleep-wake rhythm and sleep quality with endothelial function in young adults","volume":"15","author":"H Nakashima","year":"2022","journal-title":"Sleep Sci"},{"issue":"5","key":"pone.0323778.ref032","doi-asserted-by":"crossref","first-page":"791","DOI":"10.1177\/0271678X221148979","article-title":"Depression, stress and regional cerebral blood flow","volume":"43","author":"J Chiappelli","year":"2023","journal-title":"J Cereb Blood Flow Metab"},{"issue":"293","key":"pone.0323778.ref033","doi-asserted-by":"crossref","first-page":"52","DOI":"10.1080\/01621459.1961.10482090","article-title":"Multiple comparisons among means","volume":"56","author":"OJ Dunn","year":"1961","journal-title":"J Am Stat Assoc"},{"issue":"2","key":"pone.0323778.ref034","doi-asserted-by":"crossref","first-page":"80","DOI":"10.4161\/fly.19695","article-title":"A program for annotating and predicting the effects of single nucleotide polymorphisms, SnpEff: SNPs in the genome of Drosophila melanogaster strain w1118; iso-2; iso-3","volume":"6","author":"P Cingolani","year":"2012","journal-title":"Fly (Austin)"},{"issue":"7","key":"pone.0323778.ref035","doi-asserted-by":"crossref","DOI":"10.1371\/journal.pbio.1001091","article-title":"Modernizing reference genome assemblies","volume":"9","author":"DM Church","year":"2011","journal-title":"PLoS Biol"},{"issue":"6509","key":"pone.0323778.ref036","doi-asserted-by":"crossref","first-page":"1318","DOI":"10.1126\/science.aaz1776","article-title":"The GTEx consortium atlas of genetic regulatory effects across human tissues","volume":"369","author":"GTEx Consortium","year":"2020","journal-title":"Science"},{"issue":"10","key":"pone.0323778.ref037","doi-asserted-by":"crossref","first-page":"5562","DOI":"10.1073\/pnas.100118597","article-title":"A neuronal beta subunit (KCNMB4) makes the large conductance, voltage- and Ca2+-activated K+ channel resistant to charybdotoxin and iberiotoxin","volume":"97","author":"P Meera","year":"2000","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"42","key":"pone.0323778.ref038","doi-asserted-by":"crossref","first-page":"29261","DOI":"10.1074\/jbc.M114.604306","article-title":"Large conductance voltage- and Ca2+-gated potassium (BK) channel \u03b24 subunit influences sensitivity and tolerance to alcohol by altering its response to kinases","volume":"289","author":"C Vel\u00e1zquez-Marrero","year":"2014","journal-title":"J Biol Chem"},{"issue":"12","key":"pone.0323778.ref039","doi-asserted-by":"crossref","first-page":"496","DOI":"10.1016\/S0165-6147(98)01273-5","article-title":"Glucose action \u201cbeyond ionic events\u201d in the pancreatic beta cell","volume":"19","author":"T Aizawa","year":"1998","journal-title":"Trends Pharmacol Sci"},{"key":"pone.0323778.ref040","doi-asserted-by":"crossref","first-page":"1283","DOI":"10.1016\/0002-8703(91)90434-J","article-title":"Insulin resistance and compensatory hyperinsulinemia: role in hypertension, dyslipidemia, and coronary heart disease","volume":"121","author":"GM Reaven","year":"1991","journal-title":"Am Heart J"},{"key":"pone.0323778.ref041","article-title":"Phosphorylation by protein kinase A enhances delayed rectifier K+ current in rabbit vascular smooth muscle cells","volume":"268","author":"EA Aiello","year":"1995","journal-title":"Am J Physiol"},{"issue":"8","key":"pone.0323778.ref042","doi-asserted-by":"crossref","first-page":"1258","DOI":"10.1161\/CIRCRESAHA.114.303167","article-title":"Protein kinase A-phosphorylated KV1 channels in PSD95 signaling complex contribute to the resting membrane potential and diameter of cerebral arteries","volume":"114","author":"CL Moore","year":"2014","journal-title":"Circ Res"},{"issue":"1","key":"pone.0323778.ref043","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1042\/BST20130253","article-title":"cAMP signalling in the vasculature: the role of Epac (exchange protein directly activated by cAMP)","volume":"42","author":"OL Roberts","year":"2014","journal-title":"Biochem Soc Trans"},{"issue":"10","key":"pone.0323778.ref044","doi-asserted-by":"crossref","first-page":"831","DOI":"10.2174\/1567205018666211022164025","article-title":"Functional roles of neuronal nitric oxide synthase in neurodegenerative diseases and mood disorders","volume":"18","author":"CH Poon","year":"2021","journal-title":"Curr Alzheimer Res"},{"issue":"1","key":"pone.0323778.ref045","doi-asserted-by":"crossref","first-page":"101","DOI":"10.1038\/nm.3019","article-title":"Activation of calcium signaling through Trpv1 by nNOS and peroxynitrite as a key trigger of skeletal muscle hypertrophy","volume":"19","author":"N Ito","year":"2013","journal-title":"Nat Med"},{"key":"pone.0323778.ref046","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/j.brainres.2009.05.012","article-title":"PDE9A, PDE10A, and PDE11A expression in rat trigeminovascular pain signalling system","volume":"1281","author":"LS Kruse","year":"2009","journal-title":"Brain Res"},{"key":"pone.0323778.ref047","doi-asserted-by":"crossref","first-page":"174987","DOI":"10.1016\/j.ejphar.2022.174987","article-title":"A novel phosphodiesterase 9A inhibitor LW33 protects against ischemic stroke through the cGMP\/PKG\/CREB pathway","volume":"925","author":"J-Y You","year":"2022","journal-title":"Eur J Pharmacol"},{"issue":"3","key":"pone.0323778.ref048","doi-asserted-by":"crossref","first-page":"105","DOI":"10.1007\/s10194-005-0165-2","article-title":"The pathophysiology of migraine: year 2005","volume":"6","author":"MG Buzzi","year":"2005","journal-title":"J Headache Pain"},{"key":"pone.0323778.ref049","article-title":"Migraine pathophysiology","author":"PJ Goadsby","year":"2005","journal-title":"Headache"},{"issue":"3","key":"pone.0323778.ref050","doi-asserted-by":"crossref","first-page":"438","DOI":"10.1016\/j.brainresrev.2004.09.007","article-title":"Neurobiology in primary headaches","volume":"48","author":"L Edvinsson","year":"2005","journal-title":"Brain Res Brain Res Rev"},{"issue":"5","key":"pone.0323778.ref051","doi-asserted-by":"crossref","first-page":"386","DOI":"10.1038\/nrn1102","article-title":"Neurobiology of migraine","volume":"4","author":"D Pietrobon","year":"2003","journal-title":"Nat Rev Neurosci"},{"issue":"6","key":"pone.0323778.ref052","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1016\/j.phrs.2009.07.016","article-title":"Role of the PRMT-DDAH-ADMA axis in the regulation of endothelial nitric oxide production","volume":"60","author":"AJ Pope","year":"2009","journal-title":"Pharmacol Res"},{"key":"pone.0323778.ref053","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1016\/j.atherosclerosis.2015.10.101","article-title":"ADMA and arginine derivatives in relation to non-invasive vascular function in the general population","volume":"244","author":"C Baum","year":"2016","journal-title":"Atherosclerosis"},{"issue":"5","key":"pone.0323778.ref054","doi-asserted-by":"crossref","first-page":"485","DOI":"10.1097\/FJC.0000000000000218","article-title":"Tissue-specific gene expression in rat hearts and aortas in a model of vascular nitrate tolerance","volume":"65","author":"T Csont","year":"2015","journal-title":"J Cardiovasc Pharmacol"},{"issue":"3","key":"pone.0323778.ref055","doi-asserted-by":"crossref","first-page":"589","DOI":"10.1016\/j.cell.2019.08.051","article-title":"Genome-wide association studies in ancestrally diverse populations: opportunities, methods, pitfalls, and recommendations","volume":"179","author":"RE Peterson","year":"2019","journal-title":"Cell"},{"key":"pone.0323778.ref056","doi-asserted-by":"crossref","first-page":"10987","DOI":"10.1038\/srep10987","article-title":"L-citrulline for protection of endothelial function from ADMA-induced injury in porcine coronary artery","volume":"5","author":"C Xuan","year":"2015","journal-title":"Sci Rep"},{"issue":"1","key":"pone.0323778.ref057","doi-asserted-by":"crossref","first-page":"186","DOI":"10.1038\/sj.bjp.0702051","article-title":"Relaxant effects of L-citrulline in rabbit vascular smooth muscle","volume":"125","author":"E Ruiz","year":"1998","journal-title":"Br J Pharmacol"}],"container-title":["PLOS One"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0323778","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,5,29]],"date-time":"2025-05-29T17:25:15Z","timestamp":1748539515000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0323778"}},"subtitle":[],"editor":[{"given":"Asif","family":"Jan","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2025,5,29]]},"references-count":57,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2025,5,29]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0323778","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,5,29]]}}}