Abstract
Milk derivative bovine Lactoferrin (bLf), a multifunctional glycoprotein available in large quantities and recognized as safe, possesses high homology and identical functions with human Lactoferrin. There are numerous food supplements containing bLf which, however, can vary in its purity, integrity and, consequently, functionality. Here, we report on a clinical trial where bLf (100 mg two times/day) was orally administered before (Arm A) or during meals (Arm B) to pregnant women with hereditary thrombophilia and suffering from anemia of inflammation. A significant increase of the number of red blood cells (RBCs), hemoglobin (Hb), total serum iron (TSI) and serum ferritin (sFtn) levels, along with a significant decrease of interleukin-6 were detected after 30 days in Arm A, but not in Arm B, thus letting us to hypothesize that bLf inefficacy could be related to its degradation by digestive proteases. To verify this hypothesis, bLf was incubated in gastric juice collected before or after meals. An undigested or a digested profile was observed when bLf was incubated in gastric juice sampled before or after meals, respectively. These results can explain the beneficial effect observed when bLf is administered under fasting conditions, i.e. in the absence of active proteases.
References
Baker EN, Baker HM (2005) Molecular structure, binding properties and dynamics of lactoferrin. Cell Mol Life Sci 62:2531–2539. https://doi.org/10.1007/s00018-005-5368-9
Baldi A, Ioannis P, Chiara P, Eleonora F, Roubini C, Vittorio D (2005) Biological effects of milk proteins and their peptides with emphasis on those related to the gastrointestinal ecosystem. J Dairy Res 72:66–72. https://doi.org/10.1017/s002202990500110x
Brines RD, Brock JH (1983) The effect of trypsin and chymotrypsin on the in vitro antimicrobial and iron-binding properties of lactoferrin in human milk and bovine colostrum. Unusual resistance of human apolactoferrin to proteolytic digestion. Biochim Biophys Acta 759:229–235. https://doi.org/10.1016/0304-4165(83)90317-3
Cutone A, Frioni A, Berlutti F, Valenti P, Musci G, Bonaccorsi di Patti MC (2014) Lactoferrin prevents LPS-induced decrease of the iron exporter ferroportin in human monocytes/macrophages. Biometals 27:807–813. https://doi.org/10.1007/s10534-014-9742-7
Cutone A, Rosa L, Lepanto MS, Scotti MJ, Berlutti F, Bonaccorsi di Patti MC, Musci G, Valenti P (2017) Lactoferrin efficiently counteracts the inflammation-induced changes of the iron homeostasis system in macrophages. Front Immunol 8:705. https://doi.org/10.3389/fimmu.2017.00705
Cutone A, Lepanto MS, Rosa L, Scotti MJ, Rossi A, Ranucci S, De Fino I, Bragonzi A, Valenti P, Musci G, Berlutti F (2019) Aerosolized bovine lactoferrin counteracts infection, inflammation and iron dysbalance in A cystic fibrosis mouse model of Pseudomonas aeruginosa chronic lung infection. Int J Mol Sci 30:20. https://doi.org/10.3390/ijms20092128
Cutone A, Colella B, Pagliaro A, Rosa L, Lepanto MS, Bonaccorsi di Patti MC, Valenti P, Di Bartolomeo S, Musci G (2020) Native and iron-saturated bovine lactoferrin differently hinder migration in a model of human glioblastoma by reverting epithelial-to-mesenchymal transition-like process and inhibiting interleukin-6/STAT3 axis. Cell Signal 65:109461. https://doi.org/10.1016/j.cellsig.2019.109461
Dekkers BL, Kolodziejczyk E, Acquistapace S, Engmann J, Wooster TJ (2016) Impact of gastric pH profiles on the proteolytic digestion of mixed βlg-Xanthan biopolymer gels. Food Funct 7:58–68. https://doi.org/10.1039/c5fo01085c
Elbarbary HA, Abdou AM, Park EY, Nakamura Y, Mohamed HA, Sato K (2010) Novel antibacterial lactoferrin peptides generated by rennet digestion and autofocusing technique. Int Dairy J 20:646–651
Frioni A, Conte MP, Cutone A, Longhi C, Musci G, Bonaccorsi di Patti MC, Natalizi T, Marazzato M, Lepanto MS, Puddu P, Paesano R, Valenti P, Berlutti F (2014) Lactoferrin differently modulates the inflammatory response in epithelial models mimicking human inflammatory and infectious diseases. Biometals 27:843–856. https://doi.org/10.1007/s10534-014-9740-9
Gonzalez-Chavez SA, Arevalo-Gallegos S, Rascon-Cruz Q (2009) Lactoferrin: Structure, function and applications. Int J Antimicrob Agents 33:301. https://doi.org/10.1016/j.ijantimicag.2008.07.020
Groves ML (1960) The isolation of a red protein from milk. J Am Chem Soc 82:3345–3350
Hunter HN, Fulton DB, Ganz T, Vogel HJ (2002) The solution structure of human hepcidin, a peptide hormone with antimicrobial activity that is involved in iron uptake and hereditary hemochromatosis. J Biol Chem 277:37597–37603. https://doi.org/10.1074/jbc.M205305200
Jackson R, Holmes K, Phansalkar A, Rodgers GM (2008) Testing for hereditary thrombophilia: a retrospective analysis of testing referred to a national laboratory. BMC Clin Pathol 8:1–7. https://doi.org/10.1186/1472-6890-8-3
Johansson B (1960) Isolation of an iron-containing red protein from human milk. Acta Chem Scand 14:510–512
Kaandorp S, Di Nisio M, Goddijn M, Middeldorp S (2009) Aspirin or anticoagulants for treating recurrent miscarriage in women without antiphospholipid syndrome. Cochrane Database Syst Re. https://doi.org/10.1002/14651858
Krause A, Neitz S, Magert HJ, Schulz A, Forssmann WG, Schulz-Knappe P, Adermann K (2000) LEAP-1, a novel highly disulfidebonded human peptide, exhibits antimicrobial activity. FEBS Lett 480:147–150. https://doi.org/10.1016/s0014-5793(00)01920-7
Lepanto MS, Rosa L, Cutone A, Conte MP, Paesano R, Valenti P (2018) Efficacy of lactoferrin oral administration in the treatment of anemia and anemia of inflammation in pregnant and non-pregnant women: an interventional study. Front Immunol 9:2123. https://doi.org/10.3389/fimmu.2018.02123
Lepanto MS, Rosa L, Cutone A, Scotti MJ, Conte AL, Marazzato M, Zagaglia C, Longhi C, Berlutti F, Musci G, Valenti P, Conte MP (2019a) Bovine Lactoferrin pre-treatment induces intracellular killing of AIEC LF82 and reduces bacteria-induced DNA damage in differentiated human enterocytes. Int J Mol Sci 12:20. https://doi.org/10.3390/ijms20225666
Lepanto MS, Rosa L, Paesano R, Valenti P, Cutone A (2019b) Lactoferrin in aseptic and septic inflammation. Molecules 3:24. https://doi.org/10.3390/molecules24071323
Masson PL, Heremans JF, Dive C (1966) An iron-binding protein common to many external secretions. Clin Chim Acta 14:735–739
Masson PL, Heremans JF, Schonne E (1969) Lactoferrin, an iron-binding protein in neutrophilic leukocytes. J Exp Med 130:643–658
Masson PL, Heremans JF (1971) Lactoferrin in milk from different species. Comp Biochem Physiol B 39:119–129
Miralles B, Del Barrio R, Cueva C, Recio I, Amigo L (2018) Dynamic gastric digestion of a commercial whey protein concentrate†. J Sci Food Agric 98:1873–1879. https://doi.org/10.1002/jsfa.8668
Montreuil J, Tonnelat J, Mullet S (1960) Preparation and properties of lactosiderophilin (lactotransferrin) of human milk. Biochim Biophys Acta 45:413–421
Moore SA, Anderson BF, Groom CR, Haridas M, Baker EN (1997) Three-dimensional structure of diferric bovine lactoferrin at 2.8 A resolution. J Mol Biol 274:222–236. https://doi.org/10.1006/jmbi.1997.1386
Nemeth E, Rivera S, Gabayan V, Keller C, Taudorf S, Pedersen BK, Ganz T (2004) IL-6 mediates hypoferremia of inflammation by inducing the synthesis of the iron regulatory hormone hepcidin. J Clin Investig 113:1271–1276. https://doi.org/10.1172/JCI20945
Paesano R, Natalizi T, Berlutti F, Valenti P (2012a) Body iron delocalization: The serious drawback in iron disorders in both developing and developed countries. Pathog Glob Health 106:200–216. https://doi.org/10.1179/2047773212Y.0000000043
Paesano R, Pietropaoli M, Berlutti F, Valenti P (2012b) Bovine lactoferrin in preventing preterm delivery associated with sterile inflammation. Biochem Cell Biol 90:468–475. https://doi.org/10.1139/o11-060
Paesano R, Pacifici E, Benedetti S, Berlutti F, Frioni A, Polimeni A, Valenti P (2014) Safety and efficacy of lactoferrin versus ferrous sulphate in curing iron deficiency and iron deficiency anaemia in hereditary thrombophilia pregnant women: An interventional study. Biometals 27:999–1006. https://doi.org/10.1007/s10534-014-9723-x
Park CH, Valore EV, Waring AJ, Ganz T (2001) Hepcidin, a urinary antimicrobial peptide synthesized in the liver. J Biol Chem 276:7806–7810. https://doi.org/10.1074/jbc.M008922200
Piper DW, Fenton BH (1965) pH stability and activity curves of pepsin with special reference to their clinical importance. Gut 6:506–508. https://doi.org/10.1136/gut.6.5.506
Puddu P, Latorre D, Carollo M, Catizone A, Ricci G, Valenti P, Gessani S (2011) Bovine lactoferrin counteracts Toll-like receptor mediated activation signals in antigen presenting cells. PLoS ONE 6:e22504. https://doi.org/10.1371/journal.pone.0022504
Qiao B, Sugianto P, Fung E, Del-Castillo-Rueda A, Moran-Jimenez MJ, Ganz T, Nemeth E (2012) Hepcidin-induced endocytosis of ferroportin is dependent on ferroportin ubiquitination. Cell Metab 15:918–924. https://doi.org/10.1016/j.cmet.2012.03.018
Rosa L, Cutone A, Lepanto MS, Paesano R, Valenti P (2017) Lactoferrin: a natural glycoprotein involved in iron and inflammatory homeostasis. Int J Mol Sci 15:18. https://doi.org/10.3390/ijms18091985
Rosa L, Cutone A, Lepanto MS, Scotti MJ, Conte MP, Paesano R, Valenti P (2018) Physico-chemical properties influence the functions and efficacy of commercial bovine lactoferrins. Biometals 31:301–312. https://doi.org/10.1007/s10534-018-0092-8
Schubert ML, Makhlouf GM (1992) Neural, hormonal, and paracrine regulation of gastrin and acid secretion. Yale J Biol Med 65:553–560
Suzuki YA, Wong H, Ashida KY, Schryvers AB, Lönnerdal B (2008) The N1 domain of human lactoferrin is required for internalization by caco-2 cells and targeting to the nucleus. Biochemistry 47:10915–10920. https://doi.org/10.1021/bi8012164
Tang J, Pepsin A, in Neil D, Rawlings ND, Salvesen G (2013) (eds) Handbook of proteolytic enzymes, 3rd edition. Elsevier Ltd, Amsterdam, pp 27–35. ISBN 978-0-12-382219-2
Tomita MW, Bellamy M, Takase K, Yamauchi K, Wakabayashi H, Kawase K (1991) Potent antibacterial peptides generated by pepsin digestion of bovine lactoferrin. J Dairy Sci 74:4137–4142. https://doi.org/10.3168/jds.S0022-0302(91)78608-6
Valenti P, Antonini G (2005) Lactoferrin: An important host defence against microbial and viral attack. Cell Mol Life Sci 62:2576–2587. https://doi.org/10.1007/s00018-005-5372-0
Wrighting DM, Andrews NC (2006) Interleukin-6 induces hepcidin expression through STAT3. Blood 108:3204–3209. https://doi.org/10.1182/blood-2006-06-027631
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This work was granted by University of Rome La Sapienza Funds to P.V.
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PV and RP conceived and designed the clinical trial. PV, GM and LR wrote the first draft of manuscript. LR, MSL, AC, RC and RAS performed the laboratory analysis, analyzed the data, edited the manuscript, and prepared the figures and tables. All authors read and approved the final version.
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Rosa, L., Lepanto, M.S., Cutone, A. et al. Influence of oral administration mode on the efficacy of commercial bovine Lactoferrin against iron and inflammatory homeostasis disorders. Biometals 33, 159–168 (2020). https://doi.org/10.1007/s10534-020-00236-2
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DOI: https://doi.org/10.1007/s10534-020-00236-2