{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,30]],"date-time":"2026-09-30T04:24:51Z","timestamp":1790742291623,"version":"4.1.0"},"reference-count":40,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2019,12,19]],"date-time":"2019-12-19T00:00:00Z","timestamp":1576713600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2019,12,19]],"date-time":"2019-12-19T00:00:00Z","timestamp":1576713600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>There has been no report about <jats:italic>in vivo<\/jats:italic> active cholesterol-lowering dipeptide in any protein origin, despite their potential health benefits. Cattle heart protein hydrolysate ultra-filtrate (HPHU, molecular weight\u2009&lt;\u2009ca. 1,000\u2009Da peptide mixture) exhibits cholesterol-lowering activity in hypercholesterolemic rats, but the active peptide in HPHU that lowers serum cholesterol levels and its molecular mechanism are unknown. In this study, we separated and purified HPHU to identify a novel cholesterol-lowering dipeptide (phenylalanine-proline, FP) and characterized the mechanism underlying its effects <jats:italic>in vivo<\/jats:italic> and <jats:italic>in vitro<\/jats:italic>. We identified FP as an active peptide from HPHU by MALDI-TOF mass spectrometry. FP significantly decreased serum total and non-HDL cholesterol and hepatic cholesterol levels in rats. FP significantly increased serum HDL cholesterol, accompanied by a significant decrease in the atherogenic index. FP also significantly increased fecal cholesterol and acidic steroid excretion. Moreover, FP significantly decreased ATP-binding cassette transporter A1 (ABCA1) expression in the rat jejunum and reduced cholesterol absorption in Caco-2 cells. We found a novel cholesterol-lowering dipeptide FP that could improve cholesterol metabolism via the down-regulation of intestinal ABCA1. The cholesterol-lowering action induced by FP was disappeared in <jats:italic>PepT1<\/jats:italic>KO mice. FP-induced cholesterol-lowering action is mediated via PepT1 in mice.<\/jats:p>","DOI":"10.1038\/s41598-019-56031-8","type":"journal-article","created":{"date-parts":[[2019,12,19]],"date-time":"2019-12-19T11:02:52Z","timestamp":1576753372000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":43,"title":["Identification of a novel cholesterol-lowering dipeptide, phenylalanine-proline (FP), and its down-regulation of intestinal ABCA1 in hypercholesterolemic rats and Caco-2 cells"],"prefix":"10.1038","volume":"9","author":[{"given":"Arata","family":"Banno","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jilite","family":"Wang","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kenji","family":"Okada","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ryosuke","family":"Mori","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Maihemuti","family":"Mijiti","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5348-3187","authenticated-orcid":false,"given":"Satoshi","family":"Nagaoka","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2019,12,19]]},"reference":[{"key":"56031_CR1","doi-asserted-by":"publisher","first-page":"216","DOI":"10.1016\/j.atherosclerosis.2006.01.024","volume":"190","author":"T Okamura","year":"2007","unstructured":"Okamura, T. et al. The relationship between serum total cholesterol and all-cause or cause-specific mortality in a 17.3-year study of a Japanese cohort. Atherosclerosis. 190, 216\u2013223 (2007).","journal-title":"Atherosclerosis."},{"key":"56031_CR2","doi-asserted-by":"publisher","first-page":"18C","DOI":"10.1016\/0002-9149(86)91022-2","volume":"57","author":"MH Criqui","year":"1986","unstructured":"Criqui, M. H. Epidemiology of atherosclerosis: an updated overview. Am. J. Cardiol. 57, 18C\u201323C (1986).","journal-title":"Am. J. Cardiol."},{"key":"56031_CR3","doi-asserted-by":"publisher","first-page":"13","DOI":"10.1079\/BJN2003878","volume":"90","author":"W Ni","year":"2003","unstructured":"Ni, W. et al. Anti-atherogenic effect of soya and rice protein isolate, compared with casein, in apolipoprotein E-deficient mice. Br. J. Nutr. 90, 13\u201320 (2003).","journal-title":"Br. J. Nutr."},{"key":"56031_CR4","doi-asserted-by":"publisher","first-page":"836","DOI":"10.3109\/09637486.2013.804038","volume":"64","author":"L Yang","year":"2013","unstructured":"Yang, L. et al. Rice protein exerts a hypocholesterolemic effect through regulating cholesterol metabolism-related gene expression and enzyme activity in adult rats fed a cholesterol-enriched diet. Int. J. Food Sci. Nutr. 64, 836\u2013842 (2013).","journal-title":"Int. J. Food Sci. Nutr."},{"key":"56031_CR5","doi-asserted-by":"publisher","first-page":"1396","DOI":"10.1002\/jsfa.7236","volume":"96","author":"LT Tong","year":"2016","unstructured":"Tong, L. T. et al. Effects of dietary oat proteins on cholesterol metabolism of hypercholesterolaemic hamsters. J. Sci. Food Agric. 96, 1396\u20131401 (2016).","journal-title":"J. Sci. Food Agric."},{"key":"56031_CR6","first-page":"813","volume":"55","author":"S Nagaoka","year":"1991","unstructured":"Nagaoka, S., Kanamaru, Y. & Kuzuya, Y. Effects of whey protein and casein on the plasma and liver lipids in rats. Biosci. Biotechnol. Biochem. 55, 813\u2013818 (1991).","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"56031_CR7","doi-asserted-by":"publisher","first-page":"11","DOI":"10.1006\/bbrc.2001.4298","volume":"281","author":"S Nagaoka","year":"2001","unstructured":"Nagaoka, S. et al. Identification of novel hypocholesterolemic peptides derived from bovine milk \u03b2-lactoglobulin. Biochem. Biophys. Res. Commun. 281, 11\u201317 (2001).","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"56031_CR8","doi-asserted-by":"publisher","first-page":"267","DOI":"10.1007\/s11745-002-0890-6","volume":"37","author":"S Nagaoka","year":"2002","unstructured":"Nagaoka, S., Masaoka, M., Zhang, Q., Hasegawa, M. & Watanabe, K. Egg ovomucin attenuates hypercholesterolemia in rats and inhibits cholesterol absorption in Caco-2 cells. Lipids. 37, 267\u2013272 (2002).","journal-title":"Lipids."},{"key":"56031_CR9","doi-asserted-by":"publisher","first-page":"87","DOI":"10.3177\/jnsv.42.87","volume":"42","author":"L Asato","year":"1996","unstructured":"Asato, L. et al. Effects of egg white on serum cholesterol concentration in young women. J. Nutr. Sci. Vitaminol. 42, 87\u201396 (1996).","journal-title":"J. Nutr. Sci. Vitaminol."},{"key":"56031_CR10","doi-asserted-by":"publisher","first-page":"357","DOI":"10.1016\/S0021-9150(00)00456-1","volume":"151","author":"E Ros","year":"2000","unstructured":"Ros, E. Intestinal absorption of triglyceride and cholesterol. Dietary and pharmacological inhibition to reduce cardiovascular risk. Atherosclerosis. 151, 357\u2013379 (2000).","journal-title":"Atherosclerosis."},{"key":"56031_CR11","doi-asserted-by":"publisher","first-page":"1725","DOI":"10.1093\/jn\/129.9.1725","volume":"129","author":"S Nagaoka","year":"1999","unstructured":"Nagaoka, S. et al. Soy protein peptic hydrolysate with bound phospholipids decrease micellar solubility and cholesterol absorption in rats and Caco-2 cells. J. Nutr. 129, 1725\u20131730 (1999).","journal-title":"J. Nutr."},{"key":"56031_CR12","doi-asserted-by":"publisher","first-page":"977","DOI":"10.1093\/jn\/120.9.977","volume":"120","author":"M Sugano","year":"1990","unstructured":"Sugano, M. et al. Cholesterol-lowering activity of various undigested fractions of soybean protein in rats. J. Nutr. 120, 977\u2013985 (1990).","journal-title":"J. Nutr."},{"key":"56031_CR13","doi-asserted-by":"publisher","first-page":"86","DOI":"10.1007\/s11130-009-0108-1","volume":"64","author":"C Meg\u00edas","year":"2009","unstructured":"Meg\u00edas, C. et al. Sunflower protein hydrolysates reduce cholesterol micellar solubility. Plant Foods Hum Nutr. 64, 86\u201393 (2009).","journal-title":"Plant Foods Hum Nutr."},{"key":"56031_CR14","doi-asserted-by":"publisher","first-page":"607","DOI":"10.1271\/bbb.80687","volume":"73","author":"K Nakade","year":"2009","unstructured":"Nakade, K. et al. A cattle heart protein hydrolysate ameliorates hypercholesterolemia accompanied by suppression of the cholesterol absorption in rats and Caco-2 cells. Biosci. Biotechnol. Biochem. 73, 607\u2013612 (2009).","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"56031_CR15","doi-asserted-by":"publisher","first-page":"1122","DOI":"10.1021\/mp8001655","volume":"5","author":"Y Hu","year":"2008","unstructured":"Hu, Y. et al. Targeted disruption of peptide transporter Pept1 gene in mice significantly reduces dipeptide absorption in intestine. Mol. Pharm. 5, 1122\u20131130 (2008).","journal-title":"Mol. Pharm."},{"key":"56031_CR16","doi-asserted-by":"publisher","first-page":"232","DOI":"10.1271\/bbb.90730","volume":"74","author":"M Shimizu","year":"2010","unstructured":"Shimizu, M. Interaction between food substances and the intestinal epithelium. Biosci. Biotechnol. Biochem. 74, 232\u2013241 (2010).","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"56031_CR17","doi-asserted-by":"publisher","first-page":"e105073","DOI":"10.1371\/journal.pone.0105073","volume":"9","author":"Y Kashima","year":"2014","unstructured":"Kashima, Y. et al. Identification of a novel hypocholesterolemic protein, major royal jelly protein 1, derived from royal jelly. PLoS One. 9, e105073 (2014).","journal-title":"PLoS One."},{"key":"56031_CR18","doi-asserted-by":"publisher","first-page":"1738","DOI":"10.1271\/bbb.100338","volume":"74","author":"S Nagaoka","year":"2010","unstructured":"Nagaoka, S., Nakamura, S., Shibata, H. & Kanamaru, Y. Soystatin (VAWWMY), a novel bile acid-binding peptide, decreased micellar solubility and inhibited cholesterol absorption in rats. Biosci. Biotechnol. Biochem. 74, 1738\u20131741 (2010).","journal-title":"Biosci. Biotechnol. Biochem."},{"key":"56031_CR19","doi-asserted-by":"publisher","first-page":"425","DOI":"10.1038\/343425a0","volume":"343","author":"JL Goldstein","year":"1990","unstructured":"Goldstein, J. L. & Brown, M. S. Regulation of the mevalonate pathway. Nature 343, 425\u2013430 (1990).","journal-title":"Nature"},{"key":"56031_CR20","doi-asserted-by":"publisher","first-page":"239","DOI":"10.1146\/annurev-physiol-012110-142233","volume":"73,","author":"L Jia","year":"2011","unstructured":"Jia, L., Betters, J. L. & Yu, L. Niemann-pick C1-like 1 (NPC1L1) protein in intestinal and hepatic cholesterol transport. Annu. Rev. Physiol. 73,, 239\u2013259 (2011).","journal-title":"Annu. Rev. Physiol."},{"key":"56031_CR21","doi-asserted-by":"publisher","first-page":"2740","DOI":"10.1016\/j.febslet.2010.03.030","volume":"584","author":"JL Betters","year":"2010","unstructured":"Betters, J. L. & Yu, L. NPC1L1 and cholesterol transport. FEBS Lett. 584, 2740\u20132747 (2010).","journal-title":"FEBS Lett."},{"key":"56031_CR22","doi-asserted-by":"publisher","first-page":"1052","DOI":"10.1172\/JCI27352","volume":"116","author":"LR Brunham","year":"2006","unstructured":"Brunham, L. R. et al. Intestinal ABCA1 directly contributes to HDL biogenesis in vivo. J Clin Invest. 116, 1052\u20131062 (2006).","journal-title":"J Clin Invest."},{"key":"56031_CR23","doi-asserted-by":"publisher","first-page":"30432","DOI":"10.1074\/jbc.M113.501247","volume":"288","author":"J Iqbal","year":"2013","unstructured":"Iqbal, J., Parks, J. S. & Hussain, M. M. Lipid absorption defects in intestine-specific microsomal triglyceride transfer protein and ATP-binding cassette transporter A1-deficient mice. J. Biol. Chem. 288, 30432\u201330444 (2013).","journal-title":"J. Biol. Chem."},{"key":"56031_CR24","doi-asserted-by":"crossref","first-page":"28240","DOI":"10.1074\/jbc.M003337200","volume":"75","author":"P Costet","year":"2000","unstructured":"Costet, P., Luo, Y., Wang, N. & Tall, A. R. Sterol-dependent transactivation of the ABC1 promoter by the liver X receptor\/retinoid X receptor. J. Biol. Chem. 75, 28240\u201328245 (2000).","journal-title":"J. Biol. Chem."},{"key":"56031_CR25","doi-asserted-by":"publisher","first-page":"12097","DOI":"10.1073\/pnas.200367697","volume":"97","author":"A Venkateswaran","year":"2000","unstructured":"Venkateswaran, A. et al. Control of cellular cholesterol efflux by the nuclear oxysterol receptor LXR alpha. Proc. Natl. Acad. Sci. USA 97, 12097\u201312102 (2000).","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"56031_CR26","doi-asserted-by":"publisher","first-page":"1570","DOI":"10.1126\/science.1189862","volume":"328","author":"KJ Rayner","year":"2010","unstructured":"Rayner, K. J. et al. MiR-33 contributes to the regulation of cholesterol homeostasis. Science. 328, 1570\u20131573 (2010).","journal-title":"Science."},{"key":"56031_CR27","doi-asserted-by":"publisher","first-page":"12228","DOI":"10.1073\/pnas.1005191107","volume":"107","author":"TJ Marquart","year":"2010","unstructured":"Marquart, T. J., Allen, R. M., Ory, D. S. & Bald\u00e1n, A. miR-33 links SREBP-2 induction to repression of sterol transporters. Proc. Natl. Acad. Sci. USA 107, 12228\u201312232 (2010).","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"56031_CR28","doi-asserted-by":"publisher","first-page":"1566","DOI":"10.1126\/science.1189123","volume":"328","author":"SH Najafi-Shoushtari","year":"2010","unstructured":"Najafi-Shoushtari, S. H. et al. MicroRNA-33 and the SREBP host genes cooperate to control cholesterol homeostasis. Science. 328, 1566\u20131569 (2010).","journal-title":"Science."},{"key":"56031_CR29","doi-asserted-by":"publisher","first-page":"2707","DOI":"10.1161\/ATVBAHA.111.232066","volume":"31","author":"CM Ramirez","year":"2011","unstructured":"Ramirez, C. M. et al. MicroRNA-758 regulates cholesterol efflux through posttranscriptional repression of ATP-binding cassette transporter A1. Arterioscler. Thromb. Vasc. Biol. 31, 2707\u20132714 (2011).","journal-title":"Arterioscler. Thromb. Vasc. Biol."},{"key":"56031_CR30","doi-asserted-by":"publisher","first-page":"1592","DOI":"10.1161\/CIRCRESAHA.112.300626","volume":"112","author":"CM Ramirez","year":"2013","unstructured":"Ramirez, C. M. et al. Control of cholesterol metabolism and plasma high-density lipoprotein levels by microRNA-144. Circ. Res. 112, 1592\u20131601 (2013).","journal-title":"Circ. Res."},{"key":"56031_CR31","doi-asserted-by":"publisher","first-page":"1066","DOI":"10.1194\/jlr.M044925","volume":"55","author":"CM Ramirez","year":"2014","unstructured":"Ramirez, C. M. et al. RNA binding protein HuR regulates the expression of ABCA1. J. Lipid Res. 55, 1066\u20131076 (2014).","journal-title":"J. Lipid Res."},{"key":"56031_CR32","doi-asserted-by":"publisher","first-page":"2075","DOI":"10.1194\/jlr.M085985","volume":"59","author":"GM Anatharamaia","year":"2018","unstructured":"Anatharamaia, G. M. et al. Novel fatty acyl apoE mimetic peptides have increased potency to reduce plasma cholesterol in mice and macaques. J. Lipid Res. 59, 2075\u20132083 (2018).","journal-title":"J. Lipid Res."},{"key":"56031_CR33","doi-asserted-by":"publisher","first-page":"697","DOI":"10.1016\/j.bbrc.2006.11.090","volume":"352","author":"K Morikawa","year":"2007","unstructured":"Morikawa, K. et al. A novel regulatory pathway for cholesterol degradation via lactostatin. Biochem. Biophys. Res. Commun. 352, 697\u2013702 (2007).","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"56031_CR34","doi-asserted-by":"crossref","unstructured":"Nagaoka, S. Structure-function properties of hypolipidemic peptides. J. Food Biochem., e-12539 (2018).","DOI":"10.1111\/jfbc.12539"},{"key":"56031_CR35","doi-asserted-by":"publisher","first-page":"3993","DOI":"10.1021\/acs.jafc.8b01025","volume":"66","author":"S Nagaoka","year":"2018","unstructured":"Nagaoka, S. Mystery of cholesterol lowering peptide, lactostatin and soystatin. J. Agric. Food Chem. 66, 3993\u20133994 (2018).","journal-title":"J. Agric. Food Chem."},{"key":"56031_CR36","doi-asserted-by":"publisher","first-page":"469","DOI":"10.1016\/j.jff.2015.02.021","volume":"14","author":"C Lammi","year":"2015","unstructured":"Lammi, C., Zanoni, C. & Arnoldi, A. IAVPGEVA, IAVPTGVA, and LPYP, three peptides from soy glycinin, modulate cholesterol metabolism in HepG2 cells through the activation of the LDLR-SREBP2 pathway. J. Funct. Foods 14, 469\u2013478 (2015).","journal-title":"J. Funct. Foods"},{"key":"56031_CR37","doi-asserted-by":"crossref","first-page":"1583","DOI":"10.1016\/S0022-2275(20)38409-1","volume":"29","author":"I Ikeda","year":"1988","unstructured":"Ikeda, I. et al. Discrimination between cholesterol and sitosterol for absorption in rats. J. Lipid Res. 29, 1583\u20131591 (1988).","journal-title":"J. Lipid Res."},{"key":"56031_CR38","doi-asserted-by":"publisher","first-page":"28","DOI":"10.1016\/0955-2863(90)90095-3","volume":"1","author":"P Borel","year":"1990","unstructured":"Borel, P. et al. Effect of wheat bran and wheat germ on the intestinal uptake of oleic acid, monoolein, and cholesterol in the rat. J. Nutr. Biochem. 1, 28\u201333 (1990).","journal-title":"J. Nutr. Biochem."},{"key":"56031_CR39","unstructured":"Snedecor, G. W & Cochran, W. G. In: Statistical Methods, 6th ed. Ames: The Iowa State University Press. (1967)."},{"key":"56031_CR40","doi-asserted-by":"publisher","first-page":"61","DOI":"10.1214\/aos\/1176346392","volume":"12","author":"AJ Hayter","year":"1984","unstructured":"Hayter, A. J. A proof of the conjecture that the Tukey-Kramer multiple comparisons procedure is conservative. Ann. Statist. 12, 61\u201375 (1984).","journal-title":"Ann. Statist."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-56031-8.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-56031-8","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-56031-8.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,17]],"date-time":"2022-12-17T17:42:36Z","timestamp":1671298956000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-019-56031-8"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,12,19]]},"references-count":40,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2019,12]]}},"alternative-id":["56031"],"URL":"https:\/\/doi.org\/10.1038\/s41598-019-56031-8","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,12,19]]},"assertion":[{"value":"11 July 2019","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"4 December 2019","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"19 December 2019","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"19416"}}