{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,8]],"date-time":"2026-09-08T03:31:32Z","timestamp":1788838292548,"version":"build-2803163510"},"reference-count":38,"publisher":"PeerJ","license":[{"start":{"date-parts":[[2024,5,7]],"date-time":"2024-05-07T00:00:00Z","timestamp":1715040000000},"content-version":"unspecified","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Research Project of Health and Family Planning Commission of Heilongjiang Province","award":["20221212070999"],"award-info":[{"award-number":["20221212070999"]}]},{"name":"Ruikangyuan Ltd. Co.","award":["sky20200921"],"award-info":[{"award-number":["sky20200921"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"abstract":"<jats:sec>\n               <jats:title>Background<\/jats:title>\n               <jats:p>Hepatitis B virus (HBV) infection poses a major public health problem worldwide. Bovine lactoferrin (bLf) is a natural product that can inhibit HBV, but the effect of iron saturation on its resistance to HBV is unknown.<\/jats:p>\n            <\/jats:sec>\n            <jats:sec>\n               <jats:title>Aims<\/jats:title>\n               <jats:p>The purpose of this study is to investigate the impact of iron saturation of bLf against HBV.<\/jats:p>\n            <\/jats:sec>\n            <jats:sec>\n               <jats:title>Methods<\/jats:title>\n               <jats:p>HepG2 cells were cultured in DMEM high glucose containing 10% inactivated fetal calf serum, at 37\u00a0\u00b0C, in 5% CO<jats:sub>2<\/jats:sub>. MTT method was used to detect the cytotoxicity of bLf to HepG2 cells. Apo-bLf and holo-bLf were prepared from bLf. Iron saturation of these proteins was determined by atomic absorption spectrophotometry. Non-cytotoxic concentrations of candidate proteins were used in anti-HBV tests. Fluorescent quantitative polymerase chain reaction was used to detect HBV-DNA.<\/jats:p>\n            <\/jats:sec>\n            <jats:sec>\n               <jats:title>Results<\/jats:title>\n               <jats:p>The TC<jats:sub>50<\/jats:sub> and TC<jats:sub>0<\/jats:sub>of bLf were 54.570\u00a0mg\/ml and 1.997 mg\/ml, respectively. The iron saturation of bLf, apo-bLf and holo-bLf were 10.29%, 8.42% and 85.32%, respectively. In this study, four non-cytotoxic concentrations of candidate proteins (1.5, 1.0, 0.5, and 0.1 mg\/ml, respectively) were used to inhibit HBV in HepG2 cells. The results showed that 1.5 mg\/ml bLf and 0.1 mg\/ml holo-bLf effectively impaired the HBV-DNA amplification in HBV-infected HepG2 cells (<jats:italic>P<\/jats:italic> &lt; 0.05). However, apo-bLf, and Fe<jats:sup>3+<\/jats:sup> did not show the anti-HBV effects.<\/jats:p>\n            <\/jats:sec>\n            <jats:sec>\n               <jats:title>Conclusion<\/jats:title>\n               <jats:p>A total of 1.5 mg\/ml bLf and 0.1 mg\/ml holo-bLf could inhibit HBV-DNA in HepG2 cells. Complete bLf structure, appropriate concentration and iron saturation of bLf are necessary conditions for anti-HBV effects.<\/jats:p>\n            <\/jats:sec>","DOI":"10.7717\/peerj.17302","type":"journal-article","created":{"date-parts":[[2024,5,7]],"date-time":"2024-05-07T07:08:20Z","timestamp":1715065700000},"page":"e17302","source":"Crossref","is-referenced-by-count":6,"title":["Effect of iron saturation of bovine lactoferrin on the inhibition of hepatitis B virus <i>in vitro<\/i>"],"prefix":"10.7717","volume":"12","author":[{"given":"Haibo","family":"Zhou","sequence":"first","affiliation":[{"name":"College of Food Science, Northeast Agricultural University, Harbin, Heilongjiang Province, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yiwei","family":"Zhu","sequence":"additional","affiliation":[{"name":"Chongqing Food Industry Research Institute Co., Ltd., Chongqing, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ning","family":"Liu","sequence":"additional","affiliation":[{"name":"College of Food Science, Northeast Agricultural University, Harbin, Heilongjiang Province, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wencui","family":"Zhang","sequence":"additional","affiliation":[{"name":"Institute of Endemic Diseases, Harbin Center for Disease Control and Prevention, Harbin,  Heilongjiang Province, China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jianchun","family":"Han","sequence":"additional","affiliation":[{"name":"College of Food Science, Northeast Agricultural University, Harbin, Heilongjiang Province, China"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"4443","published-online":{"date-parts":[[2024,5,7]]},"reference":[{"issue":"6268","key":"10.7717\/peerj.17302\/ref-1","doi-asserted-by":"publisher","first-page":"784","DOI":"10.1038\/344784a0","article-title":"Apolactoferrin structure demonstrates ligand-induced conformational change in transferrins","volume":"344","author":"Anderson","year":"1990","journal-title":"Nature"},{"issue":"72","key":"10.7717\/peerj.17302\/ref-2","first-page":"919","article-title":"Factors influencing outcome of lamivudine in anti-HBe-positive chronic hepatitis B","volume":"53","author":"Ascione","year":"2006","journal-title":"Hepatogastroenterology"},{"issue":"6","key":"10.7717\/peerj.17302\/ref-3","doi-asserted-by":"publisher","first-page":"692","DOI":"10.1007\/s007750000157","article-title":"Metal substitution in transferrins: specific binding of cerium(IV) revealed by the crystal structure of cerium-substituted human lactoferrin","volume":"5","author":"Baker","year":"2000","journal-title":"Journal of Biological Inorganic Chemistry"},{"issue":"3","key":"10.7717\/peerj.17302\/ref-4","doi-asserted-by":"publisher","first-page":"320","DOI":"10.1139\/o11-071","article-title":"A structural perspective on lactoferrin function","volume":"90","author":"Baker","year":"2012","journal-title":"Biochemistry and Cell Biology"},{"key":"10.7717\/peerj.17302\/ref-5","doi-asserted-by":"publisher","first-page":"297","DOI":"10.1016\/j.virol.2014.01.022","article-title":"Inhibition of Mayaro virus infection by bovine lactoferrin","volume":"452\u2013453","author":"Carvalho","year":"2014","journal-title":"Virology"},{"issue":"1","key":"10.7717\/peerj.17302\/ref-6","doi-asserted-by":"publisher","first-page":"143","DOI":"10.1016\/j.virol.2008.06.017","article-title":"Bovine lactoferrin inhibits Japanese encephalitis virus by binding to heparan sulfate and receptor for low density lipoprotein","volume":"379","author":"Chien","year":"2008","journal-title":"Virology"},{"issue":"4","key":"10.7717\/peerj.17302\/ref-7","doi-asserted-by":"publisher","first-page":"1082","DOI":"10.1002\/hep.510300427","article-title":"Extended lamivudine retreatment for chronic hepatitis B: maintenance of viral suppression after discontinuation of therapy","volume":"30","author":"Dienstag","year":"1999","journal-title":"Hepatology"},{"issue":"5","key":"10.7717\/peerj.17302\/ref-8","doi-asserted-by":"publisher","first-page":"780","DOI":"10.1002\/jmv.23549","article-title":"Characterization of the anti-HBV activity of HLP1-23, a human lactoferrin-derived peptide","volume":"85","author":"Florian","year":"2013","journal-title":"Journal of Medical Virology"},{"issue":"2","key":"10.7717\/peerj.17302\/ref-9","doi-asserted-by":"publisher","first-page":"357","DOI":"10.1006\/jmbi.1993.1592","article-title":"Domain closure in lactoferrin. Two hinges produce a see-saw motion between alternative close-packed interfaces","volume":"234","author":"Gerstein","year":"1993","journal-title":"Journal of Molecular Biology"},{"issue":"24","key":"10.7717\/peerj.17302\/ref-10","doi-asserted-by":"publisher","first-page":"15655","DOI":"10.1073\/pnas.232137699","article-title":"Infection of a human hepatoma cell line by hepatitis B virus","volume":"99","author":"Gripon","year":"2002","journal-title":"Proceedings of the National Academy of Sciences of the United States of America"},{"issue":"2","key":"10.7717\/peerj.17302\/ref-11","doi-asserted-by":"publisher","first-page":"163","DOI":"10.1159\/000014021","article-title":"Protective function of proteins and lipids in human milk","volume":"74","author":"Hamosh","year":"1998","journal-title":"Biology of the Neonate"},{"issue":"2","key":"10.7717\/peerj.17302\/ref-12","doi-asserted-by":"publisher","first-page":"139","DOI":"10.2174\/1389203719666180514150921","article-title":"Lactoferrin: major physiological functions and applications","volume":"20","author":"Hao","year":"2019","journal-title":"Current Protein & Peptide Science"},{"issue":"3","key":"10.7717\/peerj.17302\/ref-13","doi-asserted-by":"publisher","first-page":"228","DOI":"10.1016\/s1386-6346(02)00088-8","article-title":"Lactoferrin inhibits hepatitis B virus infection in cultured human hepatocytes","volume":"24","author":"Hara","year":"2002","journal-title":"Hepatology Research"},{"issue":"9","key":"10.7717\/peerj.17302\/ref-14","doi-asserted-by":"publisher","first-page":"7947","DOI":"10.3390\/ijms24097947","article-title":"Iron saturation drives lactoferrin effects on oxidative stress and neurotoxicity induced by HIV-1 Tat","volume":"24","author":"Ianiro","year":"2023","journal-title":"International Journal of Molecular Sciences"},{"key":"10.7717\/peerj.17302\/ref-15","doi-asserted-by":"publisher","DOI":"10.1007\/s10534-014-9746-3","article-title":"Transcriptomic profiling of intestinal epithelial cells in response to human, bovine and commercial bovine lactoferrins","author":"Jiang","year":"2014","journal-title":"Biometals"},{"issue":"2","key":"10.7717\/peerj.17302\/ref-16","doi-asserted-by":"publisher","first-page":"61","DOI":"10.1056\/NEJM199807093390201","article-title":"A one-year trial of lamivudine for chronic hepatitis B. Asia Hepatitis Lamivudine Study Group","volume":"339","author":"Lai","year":"1998","journal-title":"New England Journal of Medicine"},{"key":"10.7717\/peerj.17302\/ref-17","doi-asserted-by":"publisher","first-page":"163","DOI":"10.1007\/978-0-387-74087-4_6","article-title":"Lactoferrin structure and functions","volume":"606","author":"Legrand","year":"2008","journal-title":"Advances in Experimental Medicine and Biology"},{"issue":"1","key":"10.7717\/peerj.17302\/ref-18","doi-asserted-by":"publisher","first-page":"19","DOI":"10.1007\/s00430-008-0100-7","article-title":"Inhibition of HBV infection by bovine lactoferrin and iron-zinc-saturated lactoferrin","volume":"198","author":"Li","year":"2009","journal-title":"Medical Microbiology and Immunology"},{"issue":"Pt 10","key":"10.7717\/peerj.17302\/ref-19","doi-asserted-by":"publisher","first-page":"2681","DOI":"10.1099\/0022-1317-75-10-2681","article-title":"In vitro infection of human hepatoma cells (HepG2) with hepatitis B virus (HBV): spontaneous selection of a stable HBV surface antigen-producing HepG2 cell line containing integrated HBV DNA sequences","volume":"75","author":"Mabit","year":"1994","journal-title":"Journal of General Virology"},{"issue":"2-3","key":"10.7717\/peerj.17302\/ref-20","doi-asserted-by":"publisher","first-page":"221","DOI":"10.1016\/0166-3542(95)00840-3","article-title":"Lactoferrin inhibits herpes simplex virus type 1 adsorption to Vero cells","volume":"29","author":"Marchetti","year":"1996","journal-title":"Antiviral Research"},{"issue":"2","key":"10.7717\/peerj.17302\/ref-21","doi-asserted-by":"publisher","first-page":"89","DOI":"10.1023\/a:1009217709851","article-title":"Metal complexes of bovine lactoferrin inhibit in vitro replication of herpes simplex virus type 1 and 2","volume":"11","author":"Marchetti","year":"1998","journal-title":"Biometals"},{"issue":"4","key":"10.7717\/peerj.17302\/ref-22","doi-asserted-by":"publisher","first-page":"199","DOI":"10.1007\/s004300050093","article-title":"Inhibition of poliovirus type 1 infection by iron-manganese- and zinc-saturated lactoferrin","volume":"187","author":"Marchetti","year":"1999","journal-title":"Medical Microbiology and Immunology"},{"issue":"1","key":"10.7717\/peerj.17302\/ref-23","doi-asserted-by":"publisher","first-page":"160","DOI":"10.1016\/j.biochi.2008.05.016","article-title":"Bovine lactoferrin and lactoferricin interfere with intracellular trafficking of Herpes simplex virus-1","volume":"91","author":"Marr","year":"2009","journal-title":"Biochimie"},{"issue":"4","key":"10.7717\/peerj.17302\/ref-24","doi-asserted-by":"publisher","first-page":"579","DOI":"10.1111\/j.1432-1033.1968.tb00484.x","article-title":"Metal-combining properties of human lactoferrin (red milk protein). 1. The involvement of bicarbonate in the reaction","volume":"6","author":"Masson","year":"1968","journal-title":"European Journal of Biochemistry"},{"key":"10.7717\/peerj.17302\/ref-25","doi-asserted-by":"publisher","first-page":"113","DOI":"10.1007\/978-1-4757-9068-9_14","article-title":"Identification and analysis of a CA(2+)-dependent lactoferrin receptor in rat liver, Lactoferrin binds to the asialoglycoprotein receptor in a galactose-independent manner","volume":"443","author":"McAbee","year":"1998","journal-title":"Advances in Experimental Medicine and Biology"},{"issue":"5","key":"10.7717\/peerj.17302\/ref-26","doi-asserted-by":"publisher","first-page":"1026","DOI":"10.1046\/j.1365-2672.2003.02071.x","article-title":"The effect of bovine lactoferrin and lactoferricin B on the ability of feline calicivirus (a norovirus surrogate) and poliovirus to infect cell cultures","volume":"95","author":"McCann","year":"2003","journal-title":"Journal of Applied Microbiology"},{"issue":"3","key":"10.7717\/peerj.17302\/ref-27","doi-asserted-by":"publisher","first-page":"258","DOI":"10.1016\/j.antiviral.2007.03.012","article-title":"The anti-papillomavirus activity of human and bovine lactoferricin","volume":"75","author":"Mistry","year":"2007","journal-title":"Antiviral Research"},{"issue":"9","key":"10.7717\/peerj.17302\/ref-28","doi-asserted-by":"publisher","first-page":"1055","DOI":"10.1016\/s1357-2725(98)00066-1","article-title":"Antiviral effect of bovine lactoferrin saturated with metal ions on early steps of human immunodeficiency virus type 1 infection","volume":"30","author":"Puddu","year":"1998","journal-title":"The International Journal of Biochemistry & Cell Biology"},{"issue":"2\u20133","key":"10.7717\/peerj.17302\/ref-29","doi-asserted-by":"publisher","first-page":"83","DOI":"10.1007\/s004300050049","article-title":"Antirotaviral activity of milk proteins: lactoferrin prevents rotavirus infection in the enterocyte-like cell line HT-29","volume":"186","author":"Superti","year":"1997","journal-title":"Medical Microbiology and Immunology"},{"issue":"4","key":"10.7717\/peerj.17302\/ref-30","doi-asserted-by":"publisher","first-page":"565","DOI":"10.1007\/s10534-017-0027-9","article-title":"Chromium(III) and iron(III) inhibits replication of DNA and RNA viruses","volume":"30","author":"Terpilowska","year":"2017","journal-title":"Biometals"},{"issue":"10170","key":"10.7717\/peerj.17302\/ref-31","doi-asserted-by":"publisher","first-page":"423","DOI":"10.1016\/S0140-6736(18)32221-9","article-title":"Enteral lactoferrin supplementation for very preterm infants: a randomised placebo-controlled trial","volume":"393","author":"The ELFIN trial investigators group","year":"2019","journal-title":"Lancet"},{"issue":"3","key":"10.7717\/peerj.17302\/ref-32","doi-asserted-by":"publisher","first-page":"493","DOI":"10.1007\/s007050050095","article-title":"Receptor-mediated entry of hepatitis B virus particles into liver cells","volume":"142","author":"Treichel","year":"1997","journal-title":"Archives of Virology"},{"issue":"8","key":"10.7717\/peerj.17302\/ref-33","doi-asserted-by":"publisher","first-page":"2101","DOI":"10.1093\/jn\/131.8.2101","article-title":"Gastric digestion of bovine lactoferrin in vivo in adults","volume":"131","author":"Troost","year":"2001","journal-title":"The Journal of Nutrition"},{"issue":"4","key":"10.7717\/peerj.17302\/ref-34","doi-asserted-by":"publisher","first-page":"1077","DOI":"10.14670\/HH-17.1077","article-title":"Immunohistochemical evidence of lactoferrin in hepatic biopsies of patients with viral or cryptogenetic chronic liver disease","volume":"17","author":"Tuccari","year":"2002","journal-title":"Histology & Histopathology"},{"issue":"3","key":"10.7717\/peerj.17302\/ref-35","doi-asserted-by":"publisher","first-page":"225","DOI":"10.1016\/s0166-3542(01)00195-4","article-title":"Antiviral activities of lactoferrin","volume":"52","author":"van der Strate","year":"2001","journal-title":"Antiviral Research"},{"key":"10.7717\/peerj.17302\/ref-36","author":"WHO","year":"2017","journal-title":"Global hepatitis report, 2017"},{"issue":"11","key":"10.7717\/peerj.17302\/ref-37","doi-asserted-by":"publisher","first-page":"2862","DOI":"10.1016\/j.bbamem.2007.07.024","article-title":"The gamma-core motif correlates with antimicrobial activity in cysteine-containing kaliocin-1 originating from transferrins","volume":"1768","author":"Yount","year":"2007","journal-title":"Biochimica et Biophysica Acta"},{"issue":"5","key":"10.7717\/peerj.17302\/ref-38","first-page":"586","article-title":"Study of inhibition effect of zinc-iron- and manganese-saturated bovine lactoferrin on hepatitis B virus DNA in vitro","volume":"37","author":"Zhou","year":"2008","journal-title":"Wei Sheng Yan Jiu"}],"container-title":["PeerJ"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/peerj.com\/articles\/17302.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/peerj.com\/articles\/17302.xml","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/peerj.com\/articles\/17302.html","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/peerj.com\/articles\/17302.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,5,7]],"date-time":"2024-05-07T07:08:31Z","timestamp":1715065711000},"score":1,"resource":{"primary":{"URL":"https:\/\/peerj.com\/articles\/17302"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,5,7]]},"references-count":38,"alternative-id":["10.7717\/peerj.17302"],"URL":"https:\/\/doi.org\/10.7717\/peerj.17302","archive":["CLOCKSS","LOCKSS","Portico"],"relation":{},"ISSN":["2167-8359"],"issn-type":[{"value":"2167-8359","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,5,7]]},"article-number":"e17302"}}