PMID: 

Biochem Biophys Res Commun. 1997 Aug 28 ;237(3):624-8. PMID: 9299415

Abstract Title: 

Apoptosis in human leukemic cells induced by lactoferricin, a bovine milk protein-derived peptide: involvement of reactive oxygen species.

Abstract: 

We examined the activity of bovine lactoferricin (Lfcin-B), a peptide derived from a bovine milk protein lactoferrin (LF-B), to induce apoptosis in THP-1 human monocytic leukemic cells. Treatment with Lfcin-B at up to 50 micrograms/ml induced cell death in THP-1 cells in dose- and time-dependent manner, showing apparent morphological changes, hypodiploid forms of genomic DNA and apoptotic DNA fragmentation, whereas LF-B was inactive even at a high dose (500 micrograms/ml). The apoptosis-inducing effect of Lfcin-B increased with reduction of serum concentration, but was inhibited by addition of Zn2+, a inhibitor of Ca2+/Mg(2+)-dependent endonucleases in a dose-dependent manner. Furthermore, Lfcin-B-induced apoptosis in THP-1 cells was completely abolished by addition of antioxidants such as N-acetyl-L-cysteine (NAC) and glutathione (GSH), but not by various cytokines and mitogen which can activate monocytic cells. In addition, THP-1 cells treated with Lfcin-B, but not LF-B, showed high levels of intracellular reactive oxygen species (ROS) from the early period (20 min) of Lfcin-B treatment. And the production of ROS by Lfcin-B was dependent upon the dose of Lfcin-B added. These results suggested that Lfcin-B, a LF-B-derived peptide, but not LF-B itself, is able to induce apoptosis in THP-1 human monocytic tumor cells, and that its apoptosis-inducing activity is related to the pathway mediated by production of the intracellular ROS and activation of Ca2+/Mg(2+)-dependent endonucleases.

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PMID: 

Biofactors. 2000 ;12(1-4):83-8. PMID: 11216510

Abstract Title: 

Prevention of colon carcinogenesis and carcinoma metastasis by orally administered bovine lactoferrin in animals.

Abstract: 

Bovine lactoferrin (bLF), a milk protein known to have bacteriostatic properties was examined for its preventive effects on colon and other organ carcinogenesis and experimental metastasis. (Experiment 1) The influence on colon carcinogenesis was investigated in male rats treated with azoxymethane (AOM), then received 2 or 0.2% bLF for 36 weeks. Significant reduction in the incidence (27% and 46% of the control, respectively) and number of adenocarcinomas of the large intestine was observed. (Experiment 2) In BALB/c mice bearing subcutaneous (s.c.) implants of colon carcinoma 26 (Co 26Lu). bLF demonstrated significant inhibition of spontaneous lung metastasis (approximately 43% of the control). Number of cytotoxic asialoGM1+ and CD8+ cells in white blood cells increased (171% and 122% of control, respectively) after treatment. Results of those experiments indicate that bLF remarkably prevents colon carcinogenesis and lung metastasis of colon carcinoma cells, possibly due to increasing cytotoxic cells in the peripheral blood.

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PMID: 

Int J Cancer. 2001 Jan 15 ;91(2):236-40. PMID: 11146451

Abstract Title: 

Orally administered bovine lactoferrin systemically inhibits VEGF(165)-mediated angiogenesis in the rat.

Abstract: 

Lactoferrin (Lf) systemically suppresses tumor growth and metastasis by unknown mechanisms. We have studied the effect of orally administered iron-unsaturated bovine Lf on angiogenesis induced by VEGF(165) and IL-1-alpha in adult rats using the mesenteric-window angiogenesis assay. VEGF(165) is a major angiogenic factor in most, if not all, tumors and other angiogenesis diseases of clinical relevance. A number of objective angiogenesis variables were analyzed using microscopic morphometry and image analysis. Lf treatment significantly inhibited the VEGF(165)-mediated response in terms of microvessel spatial extension, overall vascularity and incidence of crossover. The response to IL-1-alpha decreased significantly only in terms of microvessel crossover. In vitro, Lf exerted an antiproliferative effect on endothelial cells. To our knowledge, Lf is the first endogenous protein that has been shown to be antiangiogenic following oral administration. The oral administration of Lf thus appears to be of potential interest as an antiangiogenesis treatment modality in the clinical setting. Since tumor growth is angiogenesis dependent, the extensive therapeutic potential warrants further study to elucidate the mechanisms responsible for the angiostatic effect of Lf.

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PMID: 

Anticancer Res. 2002 May-Jun;22(3):1897-901. PMID: 12168890

Abstract Title: 

Lactoferrin-induced up-regulation of zeta (zeta) chain expression in peripheral blood T lymphocytes from cervical cancer patients.

Abstract: 

Alteration of T-cell-associated signal transduction molecules has recently been implicated in immune suppression in tumour-bearing hosts. Here we report the immunoregulatory effects of human lactoferrin (LF) on zeta-chain expression in peripheral blood T lymphocytes from cervical cancer patients and healthy donors. By quantitative flow cytometry analysis, we demonstrated that the mean zeta-chain expression was significantly higher in freshly-isolated T lymphocytes from healthy donors (69%), compared with the patients (38%). Following 3-day culture under standard conditions, zeta-chain expression in T lymphocytes from the patients increased significantly, whereas it dropped in the cells from healthy donors. Anti-CD3 MoAb as well as LF, significantly increased expression of zeta-chain in T cells both from patients and control subjects. The addition of LF to the anti-CD3 MoAb cell cultures resulted in an even higher stimulation of the zeta-chain expression. The results suggest that, in patients with cervical cancer, zeta-chain defects could be corrected by the therapeutic application of LF.

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PMID: 

Zhonghua Kou Qiang Yi Xue Za Zhi. 2010 Jan ;45(1):28-30. PMID: 20368037

Abstract Title: 

[Effects of lactoferrin on vascular endothelial growth factor and basic fibroblast growth factor expression in human Tca8113 cell line].

Abstract: 

OBJECTIVE: To investigate the effect of lactoferrin on vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) expression.METHODS: Lactoferrin at concentration of 0.006, 0.013, 0.025, 0.050 g/L and blank control groups were included in this study. The gene and protein expression of VEGF, bFGF were examined by RT-PCR and Western blotting.RESULTS: The RT-PCR and Western blotting assay showed that VEGF mRNA(0.31 +/- 0.08) and protein (0.68 +/- 0.11) in lactoferrin (0.050 g/L) group were significantly lower than in the control group (P

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PMID: 

J Pept Sci. 2008 Sep ;14(9):1032-8. PMID: 18425992

Abstract Title: 

A lactoferrin-derived peptide with cationic residues concentrated in a region of its helical structure induces necrotic cell death in a leukemic cell line (HL-60).

Abstract: 

Model studies have shown that peptides derived from the N-terminal region of bovine lactoferrin (Lf-B) exhibit antitumor activity against certain cell lines. This activity is due primarily to the peptides' apoptogenic effect. Several reports indicate that cationic residues clustered in two regions of the peptide sequence can be shuffled into one region and thereby increase cytotoxic activity, although the mechanism of this enhanced cytotoxic effect has not been clarified. In this paper, we considered several parameters that determine the mode of cell death after exposure to a native Lf-B derived peptide (Pep1, residues 17-34), and a modified peptide (mPep1) wherein the cationic residues of Pep1 are clustered in a single region of its helical structure. We found that the cytotoxic activity of mPep1 was about 9.6 fold-higher than that of Pep1 against HL-60 cells, as determined by the 3-(4,5-dimethylthiazol-2-yl)-5(3-carboxymethonyphenol)-2-(4-sulfophenyl)-2 H-tetrazolium (MTS) assay. In investigating the expression of phosphatidylserine, we observed that the native peptide (Pep1) caused both apoptotic cell death and necrotic cell death, depending on the concentration of the peptide. In contrast, the action of mPep1 was exclusively characteristic of necrotic cell death. This observation was further confirmed by agarose gel electrophoresis, in which clear ladder-like DNA bands were observed from cells exposed to Pep1, whereas DNA from cells treated with mPep1 produced a smeared pattern. We extended the study by investigating the release of mitochondrial cytochrome c into the cytosol, and the activation of caspase-3; both peptides caused the release of cytochrome c into the cytosol, and the activation of caspase-3.These results suggest that Pep1 may kill cancer cells by activating an apoptosis-inducing pathway, whereas mPep1 causes necrotic cell death by destroying cellular membrane structure notwithstanding sharing some cellular events with apoptotic cell death.

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PMID: 

J Cardiovasc Pharmacol. 2019 Dec 31. Epub 2019 Dec 31. PMID: 31895870

Abstract Title: 

Artemisinin attenuated atherosclerosis in high-fat diet-fed ApoE-/- mice by promoting macrophage autophagy via AMPK/mTOR/ULK1 pathway.

Abstract: 

Artemisinin is an endoperoxide sesquiterpene lactone from Artemisia annua L with multiple beneficial effects, including anti-inflammation, anti-oxidant and vascular protection. Recent studies have found that inflammation along with autophagy deficiency in macrophages are the possible reasons for foam cell accumulation in the intima, which leads to atherosclerotic plaque formation. The primary aim of this study was to explore the inhibiting effect of artemisinin on atherosclerosis in high-fat diet (HFD)-fed ApoE mice and investigate the probable mechanism. Artemisinin (50, 100 mg/kg, intragastric administration) treatment effectively inhibited foamy macrophage transformation and decreased atherosclerotic plaque formation in atherosclerotic mice. Moreover, artemisinin promoted AMP activated protein kinase (AMPK) activation, inhibited mammalian target of rapamycin (mTOR) and uncoordinated-51-like kinases 1 (ULK1) phosphorylation, increased LC-3II accumulation and P62 degradation, and thereby enhancing macrophage autophagy. Besides, the inhibiting effect of artemisinin on mTOR and ULK1 phosphorylation could be abrogated by AMPK knockdown, suggesting AMPK was the essential target of artemisinin on promoting macrophage autophagy. Our study indicated that artemisinin alleviated atherosclerotic lesions by accelerating macrophage autophagy via AMPK/mTOR/ULK1 pathway.

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PMID: 

Biotech Histochem. 2020 Feb ;95(2):121-128. Epub 2019 Sep 6. PMID: 32064961

Abstract Title: 

Artemisinin attenuates doxorubicin induced cardiotoxicity and hepatotoxicity in rats.

Abstract: 

We investigated the effects of artemisinin on doxorubicin (Dox) induced heart and liver pathology in rats. We divided 49 male rats into seven groups: group 1 was the untreated control. Dox was administered intraperitoneally to groups 2, 3 and 4 on day 1. Artemisinin was administered by gavage to groups 3 and 6 at a dose of 7 mg/kg, and to groups 4 and 7 at a dose of 35 mg/kg for 14 days. Group 5 was given only 0.9% NaCl orally for 14 days. At the end of the study, heart and liver samples were collected for histopathology and immunohistochemistry. Hyperemia and slight hemorrhages were observed in both livers and hearts of rats treated with Dox only. Significant increases incaspase-3, TNF-α, iNOS and NF-κB expression were observed in the myocardial cells and hepatocytes of group 2. Significant reductions in caspase-3, TNF-α, iNOS and NF-κB expression were observed in groups 3 and 4 following artemisinin treatment compared to group 2. Artemisinin may exert protective effects against Dox induced cardiotoxicity and hepatotoxicity in rats.

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PMID: 

Biometals. 2010 Jun ;23(3):507-14. Epub 2010 Apr 22. PMID: 20411301

Abstract Title: 

E2F1-directed activation of Bcl-2 is correlated with lactoferrin-induced apoptosis in Jurkat leukemia T lymphocytes.

Abstract: 

Lactoferrin (Lf) has been shown to control the proliferation of a variety of mammalian cells. Recently, we reported that human Lf induces apoptosis via a c-Jun N-terminal kinases (JNK)-associated Bcl-2 pathway that stimulates programmed cell death. In order to gain insight into the mechanism underlying Lf-triggered apoptotic features, we attempted to determine the mechanisms whereby the Lf-induced Bcl-2 family proteins exert their pro- or anti-apoptotic effects in Jurkat leukemia T lymphocytes. Treatment of the cells with high concentrations of Lf resulted in a significant reduction in in vitro growth and cell viability. As the levels of Lf increased, greater quantities of CDK6 and hyper-phosphorylated retinoblastoma protein were produced, resulting in the induction of E2F1-dependent apoptosis. Simultaneously, PARP and caspases were efficiently cleaved during Lf-induced apoptosis. The E2F1-induced apoptotic process occurred preferentially in p53-deficient Jurkat leukemia cells. Therefore, we attempted to determine whether E2F1-regulated Bcl-2 family proteins involved in the apoptotic process were relevant to Lf-induced apoptosis. We found that Lf increased the interaction of Bcl-2 with the pro-apoptotic protein Bad, whereas the total protein levels did not change significantly. Our results, collectively, suggest that Lf exploits the control mechanism of E2F1-regulated target genes or Bcl-2 family gene networks involved in the apoptotic process in Jurkat human leukemia T lymphocytes.

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PMID: 

Asian Pac J Cancer Prev. 2008 Apr-Jun;9(2):313-6. PMID: 18712982

Abstract Title: 

Lack of chronic oral toxicity of chemopreventive bovine lactoferrin in F344/DuCrj rats.

Abstract: 

Studies were undertaken to determine whether bovine lactoferrin (bLF) and related compounds, shown to prevent carcinogenesis in the colon and other organs in rats, have any toxic effects in long-term feeding studies. In experiment I, male F344/DuCrj rats received a basal diet containing 0.2% bLF for 40 weeks. No adverse findings were noted, furthermore, serum triglyceride level was significantly decreased to 72% of the control level, suggesting preventive effects against the metabolic syndrome. In experiment II, male and female F344/DuCrj rats were fed a basal diet containing 0.02, 0.2, 2.0 and 5.0% bLF, 2.0% bLF hydrolysate (bLF-H) or 0.1% lactoferricin (LFcin), a peptide derived from bLF, for 60 weeks in males and 65 weeks in females. No toxicological effects, including carcinogenicity, were evident in either sex. The results of the studies provide subjective support for safety of clinical studies of bLF for supplement use.

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