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27 pages, 9805 KiB

Open AccessArticle

Molecular Docking of Lactoferrin with Apoptosis-Related Proteins Insights into Its Anticancer Mechanism

by Lidia Esmeralda Angel-Lerma, Javier Carrillo-Campos, Luis Ignacio Siañez-Estrada, Tania Samanta Siqueiros-Cendón, Dyada Blanca León-Flores, Edward Alexander Espinoza-Sánchez, Sigifredo Arévalo-Gallegos, Blanca Flor Iglesias-Figueroa and Quintín Rascón-Cruz

Int. J. Mol. Sci. 2025, 26(5), 2023; https://doi.org/10.3390/ijms26052023 - 26 Feb 2025

Cited by 1 | Viewed by 582

Abstract

Human Lactoferrin (hLf), a multifunctional glycoprotein, has been analyzed through molecular docking to evaluate its role in apoptosis regulation and its potential as an anticancer agent. The docking results highlight XIAP (X-linked Inhibitor of Apoptosis Protein) and Caspase-3 as the most reliable targets, [...] Read more.

Human Lactoferrin (hLf), a multifunctional glycoprotein, has been analyzed through molecular docking to evaluate its role in apoptosis regulation and its potential as an anticancer agent. The docking results highlight XIAP (X-linked Inhibitor of Apoptosis Protein) and Caspase-3 as the most reliable targets, where hLf disrupts XIAP’s inhibition of Caspase-3 and Caspase-9, potentially restoring apoptotic signaling; hLf also stabilizes Caspase-3, enhancing its activation in intrinsic and extrinsic pathways. Weaker interactions were observed with Fas, Bcl-2, and Akt. hLf’s role in Fas signaling is likely due to expression upregulation rather than direct binding. In contrast, its binding to Bcl-2 may disrupt anti-apoptotic function, and its interaction with Akt suggests interference with pro-survival signaling. These findings suggest that hLf may promote apoptosis by enhancing caspase activation and modulating key apoptotic regulators, supporting its potential use in cancer treatment. However, further experimental validation is needed to confirm these interactions and their therapeutic implications. Full article

(This article belongs to the Special Issue New Insights into Lactoferrin)

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15 pages, 9972 KiB

Open AccessArticle

Lactoferrin Stimulates Chondrogenesis and Promotes Healing of the Auricular Elastic Cartilage

by Anastasiia D. Kurenkova, Natalia B. Serejnikova, Sofia A. Sheleg, Alexey L. Fayzullin, Nikolai E. Denisov, Alexandra V. Igrunkova, Elena R. Sadchikova, Artem A. Antoshin and Peter S. Timashev

Int. J. Mol. Sci. 2025, 26(5), 1956; https://doi.org/10.3390/ijms26051956 - 24 Feb 2025

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Abstract

Ear reconstruction surgeries for congenital deformities and trauma are common, highlighting the need for improved cartilage regeneration. Lactoferrin (LF), a natural and cost-effective protein, is promising due to its anti-inflammatory, antimicrobial, and prochondrogenic properties. This study investigates the effects of LF on the [...] Read more.

Ear reconstruction surgeries for congenital deformities and trauma are common, highlighting the need for improved cartilage regeneration. Lactoferrin (LF), a natural and cost-effective protein, is promising due to its anti-inflammatory, antimicrobial, and prochondrogenic properties. This study investigates the effects of LF on the viability, proliferation, and chondrogenesis of rabbit auricular chondrocytes. For in vitro studies, auricular chondrocytes were cultured for three passages, after which 3D pellets were formed. LF significantly increased chondrocyte metabolic activity by 1.5 times at doses of 10 and 500 μg/mL. At passage 3, LF at concentrations of 10 and 100 μg/mL increased cell proliferation rates by 2- and 1.5-fold, respectively. Immunohistochemical staining of the pellets demonstrated that LF at 10 μg/mL increased the amount of sex-determining region Y-Box Transcription Factor 9 (Sox9)+ cells by 30%, while at 100 μg/mL, it doubled the type II collagen deposits. For in vivo studies, a rabbit ear defect model was utilized. On post-operative day 60, the LF-treated group exhibited more mature cartilage regeneration, with a higher density of elastic fibers. By day 90 post-surgery, LF application led to the restoration of normal elastic cartilage throughout the defect. These findings suggest that LF promotes auricular chondrocytes chondrogenesis and could be beneficial for tissue engineering of the elastic cartilage. Full article

(This article belongs to the Special Issue New Insights into Lactoferrin)

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12 pages, 1482 KiB

Open AccessArticle

Bovine Lactoferrin Enhances Toll-like Receptor 7 Response in Plasmacytoid Dendritic Cells and Modulates Cellular Immunity

by Takumi Yago, Asuka Tada, Shutaro Kubo, Hirotsugu Oda, Sadahiro Iwabuchi, Miyuki Tanaka and Shinichi Hashimoto

Int. J. Mol. Sci. 2024, 25(24), 13369; https://doi.org/10.3390/ijms252413369 - 13 Dec 2024

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Abstract

Plasmacytoid dendritic cells (pDCs) express Toll-like receptor 7 (TLR7) in the endosomes, recognize viral single-stranded RNA (ssRNA), and produce significant amounts of interferon (IFN)-α. Bovine lactoferrin (LF) enhances the response of IFN regulatory factors followed by the activation of IFN-sensitive response elements located [...] Read more.

Plasmacytoid dendritic cells (pDCs) express Toll-like receptor 7 (TLR7) in the endosomes, recognize viral single-stranded RNA (ssRNA), and produce significant amounts of interferon (IFN)-α. Bovine lactoferrin (LF) enhances the response of IFN regulatory factors followed by the activation of IFN-sensitive response elements located in the promoter regions of the IFN-α gene and IFN-stimulated genes in the TLR7 reporter THP-1 cells in the presence of R-848, a TLR7 agonist. In ex vivo experiments using human peripheral blood mononuclear cells, LF enhances IFN-α levels in the supernatant in the presence of R-848. Additionally, it increases the expression of IFN-α, human leukocyte antigen (HLA)-DR, and CD86 in pDCs; HLA-DR and CD86 in myeloid dendritic cells; CD69 in CD56 dim natural killer and T killer cells; and IFN-γ in T helper type 1 and B cells in the presence of R-848. The inhibition of phagocytosis or neutralization of nucleolin, a receptor of LF, suppresses LF incorporation into pDCs. These results suggest that pDCs incorporate LF through phagocytosis or nucleolin-mediated endocytosis, and LF enhances TLR7 response in the endosome and subsequent IFN signaling pathway and activates innate and adaptive immune cells. We anticipate that LF modulates antiviral immunity against environmental ssRNA viruses and contributes to homeostasis. Full article

(This article belongs to the Special Issue New Insights into Lactoferrin)

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15 pages, 6620 KiB

Open AccessArticle

Lactoferrin Affects the Viability of Bacteria in a Biofilm and the Formation of a New Biofilm Cycle of Mannheimia haemolytica A2

by Lucero Ruiz-Mazón, Gerardo Ramírez-Rico and Mireya de la Garza

Int. J. Mol. Sci. 2024, 25(16), 8718; https://doi.org/10.3390/ijms25168718 - 9 Aug 2024

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Abstract

Respiratory diseases in ruminants are responsible for enormous economic losses for the dairy and meat industry. The main causative bacterial agent of pneumonia in ovine is Mannheimia haemolytica A2. Due to the impact of this disease, the effect of the antimicrobial protein, bovine [...] Read more.

Respiratory diseases in ruminants are responsible for enormous economic losses for the dairy and meat industry. The main causative bacterial agent of pneumonia in ovine is Mannheimia haemolytica A2. Due to the impact of this disease, the effect of the antimicrobial protein, bovine lactoferrin (bLf), against virulence factors of this bacterium has been studied. However, its effect on biofilm formation has not been reported. In this work, we evaluated the effect on different stages of the biofilm. Our results reveal a decrease in biofilm formation when bacteria were pre-incubated with bLf. However, when bLf was added at the start of biofilm formation and on mature biofilm, an increase was observed, which was visualized by greater bacterial aggregation and secretion of biofilm matrix components. Additionally, through SDS-PAGE, a remarkable band of ~80 kDa was observed when bLf was added to biofilms. Therefore, the presence of bLf on the biofilm was determined through the Western blot and Microscopy techniques. Finally, by using Live/Dead staining, we observed that most of the bacteria in a biofilm with bLf were not viable. In addition, bLf affects the formation of a new biofilm cycle. In conclusion, bLf binds to the biofilm of M. haemolytica A2 and affects the viability of bacteria and the formation a new biofilm cycle. Full article

(This article belongs to the Special Issue New Insights into Lactoferrin)

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17 pages, 3567 KiB

Open AccessArticle

Valpalf^®: A New Nutraceutical Formulation Containing Bovine Lactoferrin That Exhibits Potentiated Biological Activity

by Luigi Rosa, Giusi Ianiro, Antonella Niro, Giovanni Musci, Rosalba Paesano, Antimo Cutone and Piera Valenti

Int. J. Mol. Sci. 2024, 25(16), 8559; https://doi.org/10.3390/ijms25168559 - 6 Aug 2024

Viewed by 1503

Abstract

As a nutraceutical, bovine lactoferrin (bLf), an iron-binding glycoprotein involved in innate immunity, is gaining elevated attention for its ability to exert pleiotropic functions and to be exceptionally tolerated even at high dosages. Some of bLf’s activities, including its anti-inflammatory and antioxidant, are [...] Read more.

As a nutraceutical, bovine lactoferrin (bLf), an iron-binding glycoprotein involved in innate immunity, is gaining elevated attention for its ability to exert pleiotropic functions and to be exceptionally tolerated even at high dosages. Some of bLf’s activities, including its anti-inflammatory and antioxidant, are tightly linked to its ability to both chelate iron and enter inside the cell nucleus. Here, we present data about Valpalf^®, a new formulation containing bLf, sodium citrate, and sodium bicarbonate at a molar ratio of 10⁻³. In the present study, Valpalf^® exhibits superior iron-binding capacity, resistance to tryptic digestion, and a greater capacity to accumulate into the nucleus over time when compared to the native bLf alone. In agreement, Valpalf^® effectively reduces interleukin(IL)-6 levels in lipopolysaccharide-stimulated macrophages and modulates the expression of antioxidant enzymes, such as superoxide dismutase 1 and 2, in phorbol-12-myristate-13-acetate-stimulated monocytes. Of note, this potentiated bioactivity was corroborated in a retrospective study on the treatment of anemia of inflammation in hereditary thrombophilic pregnant and non-pregnant women, demonstrating that Valpalf^® improves hematological parameters and reduces serum IL-6 levels to a higher extent than bLf alone. Full article

(This article belongs to the Special Issue New Insights into Lactoferrin)

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13 pages, 4383 KiB

Open AccessArticle

Apo-Lactoferrin Inhibits the Proteolytic Activity of the 110 kDa Zn Metalloprotease Produced by Mannheimia haemolytica A2

by Gerardo Ramírez-Rico, Lucero Ruiz-Mazón, Magda Reyes-López, Lina Rivillas Acevedo, Jesús Serrano-Luna and Mireya de la Garza

Int. J. Mol. Sci. 2024, 25(15), 8232; https://doi.org/10.3390/ijms25158232 - 28 Jul 2024

Cited by 1 | Viewed by 1370

Abstract

Mannheimia haemolytica is the main etiological bacterial agent in ruminant respiratory disease. M. haemolytica secretes leukotoxin, lipopolysaccharides, and proteases, which may be targeted to treat infections. We recently reported the purification and in vivo detection of a 110 kDa Zn metalloprotease with collagenase [...] Read more.

Mannheimia haemolytica is the main etiological bacterial agent in ruminant respiratory disease. M. haemolytica secretes leukotoxin, lipopolysaccharides, and proteases, which may be targeted to treat infections. We recently reported the purification and in vivo detection of a 110 kDa Zn metalloprotease with collagenase activity (110-Mh metalloprotease) in a sheep with mannheimiosis, and this protease may be an important virulence factor. Due to the increase in the number of multidrug-resistant strains of M. haemolytica, new alternatives to antibiotics are being explored; one option is lactoferrin (Lf), which is a multifunctional iron-binding glycoprotein from the innate immune system of mammals. Bovine apo-lactoferrin (apo-bLf) possesses many properties, and its bactericidal and bacteriostatic effects have been highlighted. The present study was conducted to investigate whether apo-bLf inhibits the secretion and proteolytic activity of the 110-Mh metalloprotease. This enzyme was purified and sublethal doses of apo-bLf were added to cultures of M. haemolytica or co-incubated with the 110-Mh metalloprotease. The collagenase activity was evaluated using zymography and azocoll assays. Our results showed that apo-bLf inhibited the secretion and activity of the 110-Mh metalloprotease. Molecular docking and overlay assays showed that apo-bLf bound near the active site of the 110-Mh metalloprotease, which affected its enzymatic activity. Full article

(This article belongs to the Special Issue New Insights into Lactoferrin)

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15 pages, 3990 KiB

Open AccessArticle

Production of Bioactive Porcine Lactoferrin through a Novel Glucose-Inducible Expression System in Pichia pastoris: Unveiling Antimicrobial and Anticancer Functionalities

by Chih-Ching Yen, Pei-Ying Wu, Huan Ou-Yang, Hsiao-Ling Chen, Kowit-Yu Chong, Ro-Lin Chang and Chuan-Mu Chen

Int. J. Mol. Sci. 2024, 25(3), 1818; https://doi.org/10.3390/ijms25031818 - 2 Feb 2024

Cited by 5 | Viewed by 3630

Abstract

Lactoferrin (LF) stands as one of the extensively investigated iron-binding glycoproteins within milk, exhibiting diverse biological functionalities. The global demand for LF has experienced consistent growth. Biotechnological strategies aimed at enhancing LF productivity through microbial expression systems offer substantial cost-effective advantages and exhibit [...] Read more.

Lactoferrin (LF) stands as one of the extensively investigated iron-binding glycoproteins within milk, exhibiting diverse biological functionalities. The global demand for LF has experienced consistent growth. Biotechnological strategies aimed at enhancing LF productivity through microbial expression systems offer substantial cost-effective advantages and exhibit fewer constraints compared to traditional animal bioreactor technologies. This study devised a novel recombinant plasmid, wherein the AOX1 promoter was replaced with a glucose-inducible G1 promoter (P_G1) to govern the expression of recombinant porcine LF (rpLF) in Pichia pastoris GS115. High-copy-number P_G1-rpLF yeast clones were meticulously selected, and subsequent induction with 0.05 g/L glucose demonstrated robust secretion of rpLF. Scaling up production transpired in a 5 L fermenter, yielding an estimated rpLF productivity of approximately 2.8 g/L by the conclusion of glycerol-fed fermentation. A three-step purification process involving tangential-flow ultrafiltration yielded approximately 6.55 g of rpLF crude (approximately 85% purity). Notably, exceptional purity of rpLF was achieved through sequential heparin and size-exclusion column purification. Comparatively, the present glucose-inducible system outperformed our previous methanol-induced system, which yielded a level of 87 mg/L of extracellular rpLF secretion. Furthermore, yeast-produced rpLF demonstrated affinity for ferric ions (Fe³⁺) and exhibited growth inhibition against various pathogenic microbes (E. coli, S. aureus, and C. albicans) and human cancer cells (A549, MDA-MB-231, and Hep3B), similar to commercial bovine LF (bLF). Intriguingly, the hydrolysate of rpLF (rpLFH) manifested heightened antimicrobial and anticancer effects compared to its intact form. In conclusion, this study presents an efficient glucose-inducible yeast expression system for large-scale production and purification of active rpLF protein with the potential for veterinary or medical applications. Full article

(This article belongs to the Special Issue New Insights into Lactoferrin)

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Review

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17 pages, 1496 KiB

Open AccessReview

Antioxidant Potential of Lactoferrin and Its Protective Effect on Health: An Overview

by Quintín Rascón-Cruz, Tania Samanta Siqueiros-Cendón, Luis Ignacio Siañez-Estrada, Celina María Villaseñor-Rivera, Lidia Esmeralda Ángel-Lerma, Joel Arturo Olivas-Espino, Dyada Blanca León-Flores, Edward Alexander Espinoza-Sánchez, Sigifredo Arévalo-Gallegos and Blanca Flor Iglesias-Figueroa

Int. J. Mol. Sci. 2025, 26(1), 125; https://doi.org/10.3390/ijms26010125 - 26 Dec 2024

Cited by 7 | Viewed by 1566

Abstract

Chronic diseases, including cardiovascular and neurodegenerative diseases and cancer, are significant global health challenges. Oxidative stress, characterized by an imbalance between reactive oxygen species (ROS) production and antioxidant defenses, is a critical factor in the progression of these pathologies. Lactoferrin (Lf), a multifunctional [...] Read more.

Chronic diseases, including cardiovascular and neurodegenerative diseases and cancer, are significant global health challenges. Oxidative stress, characterized by an imbalance between reactive oxygen species (ROS) production and antioxidant defenses, is a critical factor in the progression of these pathologies. Lactoferrin (Lf), a multifunctional iron-binding glycoprotein, has emerged as a promising therapeutic agent due to its potent antioxidant, anti-inflammatory, and iron-regulating properties. Lf plays a pivotal role in iron homeostasis by chelating iron, modulating its cellular uptake, and reducing ROS production, thereby mitigating oxidative stress-related tissue damage. Lf also demonstrates neuroprotective potential in diseases like Parkinson’s and Alzheimer’s, where it alleviates oxidative damage, regulates iron metabolism, and enhances antioxidant defenses. Furthermore, its ability to enhance endogenous antioxidant mechanisms, such as superoxide dismutase and glutathione peroxidase, underscores its systemic protective effects. Lf’s anti-inflammatory and antimicrobial activities also contribute to its broad-spectrum protective role in chronic diseases. This review consolidates evidence of Lf’s mechanisms in mitigating oxidative stress and highlights its therapeutic potential as a versatile molecule for preventing and managing chronic conditions linked to oxidative damage. Full article

(This article belongs to the Special Issue New Insights into Lactoferrin)

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