In previous studies we showed that colostrinin (CLN), a complex of proline-rich polypeptides derived from ovine colostrum, induces mitogenic stimulation, as well as a variety of cytokines in human peripheral blood leukocytes, and possesses antioxidant activity in pheochromocytoma (PC12) cells. In this study we investigated the effects of CLN on 4-hydroxynonenal (4HNE)-mediated adduct formation, generation of reactive oxygen species (ROS), glutathione (GSH) metabolism, and the modification of signal transduction cascade that leads to activation of c-Jun N-terminal kinase (JNK) in PC12 cells. Here we demonstrate that CLN (1) reduced the abundance of 4HNE-protein adducts, as shown by fluorescent microscopy and Western blot analysis; (2) reduced intracellular levels of ROS, as shown by a decrease in 2′,7′-dichlorodihydro-fluorescein-mediated fluorescence; (3) inhibited 4HNE-mediated GSH depletion, as determined fluorimetrically; and (4) inhibited 4HNE-induced activation of JNKs. Together, these findings suggest that CLN appears to down-regulate 4HNE-mediated lipid peroxidation and its product-induced signaling that otherwise may lead to pathological changes at the cellular and organ level. These findings also suggest further that CLN could be useful in the treatment of diseases such as Alzheimer’s, as well as those in which ROS are implicated in pathogenesis.
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Boldogh I., Hughes T. K., Georgiades J. A., and Stanton J. (2000) Antioxidant and nerve differentiating activity of colostrinine and related peptides, in First International Symposium on Colostrinin Georgiades J., ed. Lodz, Poland.
Boldogh I., Hughes T. K., Georgiades J. A., and Stanton J. (2001) Antioxidant and cell-differentiating activity of colostrinin and its component peptides (CCP) in cell culture. Yearbook of Psychogeriatry 4, 57–65.
Brown A. J., Leong S. L., Dean R. T., and Jessup W. (1997) 7-Hydroperoxycholesterol and its products in oxidized low density lipoprotein and human atherosclerotic plaque. J. Lipid Res. 38, 1730–1745.
Bruce-Keller A. J., Li Y. J., Lovell M. A., Kraemer P. J., Gary D. S., Brown R. R., et al. (1998) 4-Hydroxynonenal, a product of lipid peroxidation, damages cholinergic neurons and impairs visuospatial memory in rats. J. Neuropathol. Exp. Neurol. 57, 257–267.
Buettner G. R. (1993) The pecking order of free radicals and antioxidants: lipid peroxidation, alpha-tocopherol, and ascorbate. Arch. Biochem. Biophys. 300, 535–543.
Cadenas E. and Davies K. J. (2000) Mitochondrial free radical generation, oxidative stress, and aging. Free Radic. Biol. Med. 29, 222–230.
Camandola S., Poli G., and Mattson M. P. (2000) The lipid peroxidation product 4-hydroxy-2,3-nonenal inhibits constitutive and inducible activity of nuclear factor kappa B in neurons. Brain Res. Mol. Brain Res. 85, 53–60.
Cheng J. Z., Singhal S. S., Saini M., Singhal J., Piper J. T., Van Kuijk F. J., et al. (1999) Effects of mGST A4 trans-fection on 4-hydroxynonenal-mediated apoptosis and differentiation of K562 human erythroleukemia cells. Arch. Biochem. Biophys. 372, 29–36.
Davies M. J. and Truscott R. J. (2001) Photo-oxidation of proteins and its role in cataractogenesis. J. Photochem. Photobiol. B 63, 114–125.
Davis W. Jr., Ronai Z., and Tew K. D. (2001) Cellular thiols and reactive oxygen species in drug-induced apoptosis. J. Pharmacol. Exp. Ther. 296, 1–6.
de Zwart L. L., Meerman J. H., Commandeur J. N., and Vermeulen N. P. (1999) Biomarkers of free radical damage applications in experimental animals and in humans. Free Radic. Biol. Med. 26, 202–226.
Esterbauer H., Schaur R. J., and Zollner H. (1991) Chemistry and biochemistry of 4-hydroxynonenal, malonaldehyde and related aldehydes. Free Radic. Biol. Med. 11, 81–128.
Evan G. and Littlewood T. (1998) A matter of life and cell death. Science 281, 1317–1322.
Finkel T. Holbrook N. J. (2000) Oxidants, oxidative stress and the biology of ageing. Nature 408, 239–247.
Friguet B., Bulteau A. L., Chondrogianni N., Conconi M., and Petropoulos I. (2000) Protein degradation by the proteasome and its implications in aging. Ann. NY Acad. Sci. 908, 143–154.
Gardner J. L. and Gallagher E. P. (2001) Development of a peptide antibody specific to human glutathione S-transferase alpha 4-4 (hGSTA4-4) reveals preferential localization in human liver mitochondria. Arch. Biochem. Biophys. 390, 19–27.
Hainaut P. and Milner J. (1993) Redox modulation of p53 conformation and sequence-specific DNA binding in vitro. Cancer Res. 53, 4469–4473.
Han S. I., Oh S. Y., Woo S. H., Kim K. H., Kim J. H., Kim H. D., and Kang H. S. (2001) Implication of a small GTPase Rac1 in the activation of c-Jun N-terminal kinase and heat shock factor in response to heat shock. J. Biol. Chem. 276, 1889–1895.
Hughes A. L., Gollapudi L., Sladek T. L., and Neet K. E. (2000) Mediation of nerve growth factor-driven cell cycle arrest in PC12 cells by p53. Simultaneous differentiation and proliferation subsequent to p53 functional inactivation. J. Biol. Chem. 275, 37829–37837.
Inglot A. D., Gelder F., and Georgiades J. A. (1998) Tumor-associated antigens are cytokine inducers and hypo-reactivity factors to the immune system. Biotherapy 11, 27–37.
Janusz, M. and Lisowski J. (1993) Proline-rich polypeptide (PRP)—an immunomodulatory peptide from ovine colostrum. Arch. Immunol. Ther. Exp. 41, 275–279.
Janusz M., Lisowski J., and Franek F. (1974) Isolation and characterization of a proline-rich polypeptide from ovine colostrum. FEBS Lett. 49, 276–279.
Janusz M., Staroscik K., Zimecki M., Wieczorek Z., and Lisowski J. (1981) Chemical and physical characterization of a proline-rich polypeptide from sheep colostrum. Biochem. J. 199, 9–15.
Keller J. N., Kindy M. S., Holtsberg F. W., St Clair D. K., Yen H. C., Germeyer A., S., et al. (1998) Mitochondrial managanese superoxide dismutase prevents neural apoptosis and reduces ischemic brain injury: suppression of peroxynitrite production, lipid peroxidation, and mitochondrial dysfunction. J. Neurosci. 18, 687–697.
Kong A. N., Yu R., Chen C., Mandlekar S., and Primiano T. (2000) Signal transduction events elicited by natural products: role of MAPK and caspase pathways in homeostatic response and induction of apoptosis. Arch. Pharmacol Res. 23, 1–16.
Kruman I., Bruce-Keller A. J., Bredesen D., Waeg G., and Mattson M. P. (1997) Evidence that 4-hydroxynonenal mediates oxidative stress-induced neuronal apoptosis. J. Neurosci. 17, 5089–5100.
Kruzel M., Janusz M., Lisowski J., Fischleigh, R., and Georgiades J. A. (2001) Towards an understanding of biological role of colostrinin peptides. J. Mol. Neurosci. 17, 115–125.
Lafon-Cazal M., Culcasi M., Gaven F., Pietri S., and Bockaert J. (1993) Nitric oxide, superoxide and peroxynitrite: putative mediators of NMDA-induced cell death in cerebellar granule cells. Neuropharmacology 32, 1259–1266.
LeBel C. P., Ischiropoulos H., and Bondy S. C. (1992) Evaluation of the probe 2′,7′-dichlorofluorescin as an indicator of reactive oxygen species formation and oxidative stress. Chem. Res. Toxicol. 5, 227–231.
Leonarduzzi G., Arkan M. C., Basaga H., Chiarpotto E., Sevanian A., and Poli G. (2000) Lipid oxidation products in cell signaling. Free Radic. Biol. Med. 28, 1370–1378.
Lovell M. A., Ehmann W. D., Mattson M. P., and Markesbery W. R. (1997) Elevated 4-hydroxynonenal in ventricular fluid in Alzheimer’s disease. Neurobiol. Aging 18, 457–461.
Markesbery W. R. (1997) Oxidative stress hypothesis in Alzheimer’s disease. Free Radic. Biol. Med. 23, 137–147.
Markesbery W. R. and Carney J. M. (1999) Oxidative alterations in Alzheimer’s disease. Brain Pathol. 9, 133–146.
Markesbery W. R. and Lovell M. A. (1998) Four-hydroxynonenal, a product of lipid peroxidation, is increased in the brain in Alzheimer’s disease. Neurobiol. Aging 19, 33–36.
Mattson M. P. and Furukawa K. (1997) Alzheimer’s disease. Short precursor shortens memory. Nature 387, 457–458.
Mecocci P., MacGarvey U., and Beal M. F. (1994) Oxidative damage to mitochondrial DNA is increased in Alzheimer’s disease. Ann. Neurol. 36, 747–751.
Montine T. J., Markesbery W. R., Morrow J. D., and Roberts L. J. II (1998) Cerebrospinal fluid F2-isoprostane levels are increased in Alzheimer’s disease. Ann. Neurol. 44, 410–413.
Nakamura H., Nakamura K., and Yodoi J. (1997) Redox regulation of cellular activation. Annu. Rev. Immunol. 15, 351–369.
Page S., Fischer C., Baumgartner B., Haas M., Kreusel U., Loidl G., et al. (1999) 4-Hydroxynonenal prevents NF-kappaB activation and tumor necrosis factor expression by inhibiting IkappaB phosphorylation and subsequent proteolysis. J. Biol. Chem. 274, 11611–11618.
Parola M., Robino G., Marra F., Pinzani M., Bellomo G., Leonarduzzi G., et al. (1998) HNE interacts directly with JNK isoforms in human hepatic stellate cells. J. Clin. Invest. 102, 1942–1950.
Perkins A. J., Hendrie H. C., Callahan C. M., Gao S., Unverzagt F. W., Xu Y., et al. (1999) Association of antioxidants with memory in a multiethnic elderly sample using the Third National Health and Nutrition Examination Survey. Am. J. Epidemiol. 150, 37–44.
Perrig W. J., Perrig P., and Stahelin H. B. (1997) The relation between antioxidants and memory performance in the old and very old. J. Am. Geriatr. Soc. 45, 718–724.
Poli G. and Schaur R. J. (2000) 4-Hydroxynonenal in the pathomechanisms of oxidative stress. International Union of Biochemistry and Molecular Biology Life 50, 315–321.
Prasad M. R., Lovell M. A., Yatin M., Dhillon H., and Markesbery W. R. (1998) Regional membrane phospholipid alterations in Alzheimer’s disease. Neurochem. Res. 23, 81–88.
Rivas-Arancibia S., Vazquez-Sandoval R., Gonzalez-Kladiano D., Schneider-Rivas S., and Lechuga-Guerrero A. (1998) Effects of ozone exposure in rats on memory and levels of brain and pulmonary superoxide dismutase. Environ. Res. 76, 33–39.
Ross J. S., Stagliano N. E., Donovan M. J., Breitbart R. E., and Ginsburg G. S. (2001) Atherosclerosis: a cancer of the blood vessels? Am. J. Clin. Pathol. 116 (Suppl.), S97–107.
Rusnak F. and Reiter T. (2000) Sensing electrons: protein phosphatase redox regulation. Trends Biochem. Sci. 25, 527–529.
Sano M., Ernesto C., Thomas R. G., Klauber M. R., Schafer K., Grundman M., (1997) A controlled trial of selegiline, alpha-tocopherol, or both as treatment for Alzheimer’s disease. The Alzheimer’s Disease Cooperative Study. N. Engl. J. Med. 336, 1216–1222.
Sayre L. M., Zelasko D. A., Harris P. L., Perry G., Salomon R. G., and Smith M. A. (1997) 4-Hydroxynonenal-derived advanced lipid peroxidation end products are increased in Alzheimer’s disease. J. Neurochem. 68, 2092–2097.
Senft A. P., Dalton T. P., and Shertzer H. G. (2000) Determining glutathione and glutathione disulfide using the fluorescence probe o-phthalaldehyde. Anal. Biochem. 280, 80–86.
Sinclair A. J., Bayer A. J., Johnston J., Warner C., and Maxwell S. R. (1998) Altered plasma antioxidant status in subjects with Alzheimer’s disease and vascular dementia. Int. J. Geriatr. Psychiatry 13, 840–845.
Stanton J., Boldogh I., Georgiades J. A., and Hughes T. K. (2001) Induction of proliferation and cytokines by colostrinin and component proline rich peptides in human peripheral blood leukocytes. Yearbook of Psychogeriatry 4, 67–75.
Uchida K. and Stadtman E. R. (1992) Modification of histidine residues in proteins by reaction with 4-hydroxynonenal. Proc. Natl. Acad. Sci. USA 89, 4544–4548.
Vaglini F., Pardini C., Viaggi C., and Corsini G. U. (2001) Cytochrome P450 and parkinsonism: protective role of CYP2E1. Funct. Neurol. 16, 107–112.
Woods D. B. and Vousden K. H. (2001) Regulation of p53 function. Exp. Cell Res. 264, 56–66.
Yoritaka A., Hattori N., Uchida K., Tanaka M., Stadtman E. R., and Mizuno Y. (1996) Immunohistochemical detection of 4-hydroxynonenal protein adducts in Parkinson disease. Proc. Natl. Acad. Sci. USA 93, 2696–2701.
Zimecki M. and Pierce A. (1984) Immunotropic properties of fractions isolated from human milk. Arch. Immunol. Ther. Exp. 32, 203–209.
Zimecki M., Lisowski J., Hraba T., Wieczorek Z., Janusz M., and Staroscik K. (1984a) The effect of a proline-rich polypeptide (PRP) on the humoral immune response. I. Distinct effect of PRP on the T cell properties of mouse glass-nonadherent (NAT) and glass-adherent (GAT) thymocytes in thymectomized mice. Arch. Immunol. Ther. Exp. 32, 191–196.
Zimecki M., Lisowski J., Hraba T., Wieczorek Z., Janusz M., and Staroscik K. (1984b) The effect of a proline-rich polypeptide (PRP) on the humoral immune response. II. PRP induces differentiation of helper cells from glassnonadherent thymocytes (NAT) and suppressor cells from glass-adherent thymocytes (GAT). Arch. Immunol. Ther. Exp. 32, 197–201.
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Boldogh, I., Liebenthal, D., Hughes, T.K. et al. Modulation of 4HNE-mediated signaling by proline-rich peptides from ovine colostrum. J Mol Neurosci 20, 125–134 (2003). https://doi.org/10.1385/JMN:20:2:125