Nucleo CMP Forte<sup>®</sup> is a nucleotide-based drug consisting of cytidinemonophosphate, uridinemonophosphate, uridin-ediphosphate and uridinetriphosphate. It has been prescribed for peripheral nervous system disorders, such as lum-bosciatalgia, diabetic or alcoholic polyneuropathy, or trigeminal neuralgia. Its effects on brain pathologies has re-ceived little attention. We examined its neuroprotective effects on cell toxicity induced by glutamate excitotoxicity or by 1-methyl-4-phenyl-pyridinium (MPP<sup>+</sup>), an in vitro cell model of Parkinson’s disease. We used the human dopaminergic cell line SH-SY5Y and a primary culture of rat cortical cells pre-treated with the drug for 24 hours and then exposed to MPP<sup>+</sup> or glutamate at a range of concentrations. Cell viability was measured at different times. Nucleo CMP Forte<sup>®</sup> pre-treatment significantly increased the rate of cell division in SH-SY5Y cells, as well as the synthesis of triglycerides and phospholipids. More interestingly, drug pre-treatment significantly reduced MPP<sup>+</sup>- and glutamate-induced cell death in SH-SY5Y cells and in rat cortical cells. These results indicate that the nucleotides included in Nucleo CMP Forte<sup>®</sup> are promising therapeutic molecules for the prevention of neuronal death in brain caused by focal ischemia, Parkinson’s disease or other neurodegenerative pathologies.
D.W. Choi, S.M. Rothman, “The role of glutamate neurotoxicity in hypoxic-ischemic neuronal death” Neurosci, Vol. 13, 1990, pp. 171-182.
R.L. Hayes, L.W. Jenkins, BG. Lyeth, “Neurotransmitter-mediated mechanisms of traumatic brain injury: acetylcholine and excitatory amino acids” J TraumaSuppl, Vol. 1, 1992, pp. S173-S187.
S.M. Rothman, J.W. Olney, “Excitotoxicity and the NMDA receptor--still lethal after eight years” Trends Neurosci, Vol. 10,1995, pp. 299-302.
B.B. Meldrum, J. Garthwaita, “Excitatory amino acid neurotoxicity and neurodegenerative disease” Trends PharmacolSci, Vol. 11, No. 9,1990, pp. 379-387.
D.W. Choi, “Enhancement of outward potassium current may participate in beta-amyloid peptide-induced cortical neuronal death” Neuron, Vol. 1, 1998, pp. 623-634.
Y. Zhang, B.R. Bhavnani, “Glutamate-induced apoptosis in neuronal cells is mediated via caspase-dependent and independent mechanisms involving calpain and caspase-3 proteases as well as apoptosis inducing factor (AIF) and this process is inhibited by equine estrogens” BMC Neurosci, Vol.7, 2006,pp. 49.
M.J. Zigmond, E.M. Stricker, “Animal models of parkinsonism using selective neurotoxins: clinical and basic implications” Int Rev Neurobiol, Vol. 31,1989, pp. 1-79.
O. Eberhardt, J.B. Schulz, “Apoptotic mechanisms and antiapoptotic therapy in the MPTP model of Parkinson's disease” ToxicolLett, Vol. 139,2003,pp. 135-151.
V. Gallai, G. Mazzotta, S. Montesi, P. Sarchelli, F. Del Gatto, “Effects of uridine in the treatment of diabetic neuropathy: an electrophysiological study”ActaNeurol Scand, Vol. 86, No.1, 1992, pp. 3-7.
L. Cartier, J.L. Castillo, R. Verdugo, “Effect of the Nucleus CMP forte in 46 patients with progressive spastic paraparesis. Randomized and blind study” Rev Med Chil, Vol. 124, No. 5,1996, pp. 583-587.
C. Kretschmar, S. Kaumeier, W. Hasse, “Medicamentous therapy of alcoholic polyneuropathy. Randomized double-blind study comparing 2 vitamin B preparations and a nucleotide preparation” Fortschr Med, Vol. 114, No. 32, 1996,pp.439-443.
B. Watting, G. Schalow, F. Heydenreich, R. Warzok, J. Cervós-Navarro “Enhancement of nerve fibre regeneration by nucleotides after peripheral nerve crush damage” Electrophysiologic and morphometric investigations. Arzeinmittelforshung, Vol. 42, 1992, pp.1075-1078.
W. Araki and R.J. Wurtman, “How is membrane phospholipid biosynthesis controlled in neural tissues?”J Neurosci Res, Vol. 51, No. 6,1998, pp. 667-674.
J.R. Marszalek, H.F. Lodish, “Docosahexaenoic acid, fatty acid-interacting proteins, and neuronal function: breastmilk and fish are good for you” Annu Rev Cell Dev Biol, Vol. 21, 2005, pp. 633-657.
S.I. Rapoport, “In vivo fatty acid incorporation into brain phosholipids in relation to plasma availability, signal transduction and membrane remodeling” J Mol Neurosci, Vol. 16, 2001, pp. 234-261.
E.S. Haugaard, K.B. Frantz, N. Haugaard, “Effect of uridine on cellular UTP and glycogen synthesis in skeletal muscle: stimulation of UTP formation by insulin” Proc NatlAcadSci USA, Vol. 74, No. 6,1977, pp. 2339-2342.
N.N. Suzuki, K. Koizumi, M. Fukushima, A. Matasuda, F. Inagaki, “Structural basis for the specificity, catalysis, and regulation of human uridine-cytidinekinase”Structure, Vol. 12,2004, pp. 751-764.
A. Gella, J. Ponce, R. Cussó, N. Durany, “Effect of the nucleotides CMP and UMP on exhaustion in exercise rats”J Physiol Biochem, Vol. 64, No.1,2008, pp. 9-17.
M. Okada, T. Nakagawa, M. Minami, M. Satoh, “Analgesic effects of intrathecal administration of P2Y nucleotide receptor agonists UTP and UDP in normal and neuropathic pain model rats”J Pharmacol Exp Ther, Vol. 303, No. 1,2002, pp. 66-73.
LA. Teather, RJ. Wurtman, “Chronic administration of UMP ameliorates the impairment of hippocampal-dependent memory in impoverished rats”J Nutr,Vol.136, No.11,2006, pp. 2834-2837.
T. Sakamoto, M. Cansev, RJ. Wurtman, “Oral supplementation with docosahexaenoic acid and uridine-5'-monophosphate increases dendritic spine density in adult gerbil hippocampus”Brain Res, Vol. 28, 2007, pp.50-59.
L. Wang, MA. Albrecht, R.J. Wurtman, “Dietary supplementation with uridine-5'-monophosphate (UMP), a membrane phosphatide precursor, increases acetylcholine level and release in striatum of aged rat”Brain Res, Vol. 1133, No. 1,2007, pp. 42-48.
M.L. Tian, Z. Zou, H.B. Yuan, CC. Wang, Q.F. Zhu, H.T. Xu, X. Gao, XY. Shi, “Uridine 5'-triphosphate (UTP) protects against cerebral ischemia reperfusion injury in rats”Neurosci Lett, Vol. 465, No. 1,2009, pp. 55-60.
M.B Hansen, SE. Nielsen, K. Berg, “Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill” J Immunol Methods, Vol. 119, No. 2,1989, pp. 203-210.
D. Lecca, S. Ceruti, “Uracil nucleotides: from metabolic intermediates to neuroprotection and neuroinflammation” Biochemical Pharmacology, Vol. 75,2008, pp. 1869-1881.
W. Fischer, U. Krügel, “P2Y receptors: focus on structural, pharmacological and functional aspects in the brain”Curr Med Chem, Vol. 14, No. 23,2007, pp. 2429-1455.
A. Brunschweiger, CE. Müller, “P2 receptors activated by uracil nucleotides--an update.” Curr Med Chem, Vol.13, No. 3, 2006, pp. 289-312.
H.R. Xie, LS. Hu, G.Y. Li, “SH-SY5Y human neuroblastoma cell line: in vitro cell model of dopaminergic neurons in Parkinson's disease”Chin Med J, Vol. 123, No. 8, 2010, pp. 1086-1092.
A. Cuppelo, A. Dierich, P. Mandel, M. Wintzerith, “In vitro protein synthesis activity of poly(A) RNA from neuroblastoma cells” Neurochem Res, Vol. 5,1980, pp. 271-279.
T. Ming-Tzeu, L. Shue-Feno, W. Chuan-Chawng, C. Chin-Sung, C.H. Chi-Tso, L. Chih-Chung, Y. Chuen-Mang, “P2Y2 receptor mediated proliferation of C6 glioma cells via activation of Ras/Raf/MEK/MAPK pathway” Br J Pharmacol, Vol. 129, No. 7,2000, pp. 1481-1489.
J.T. Nearg, Y. Kang, Y. Bu, E. Yu, K. Akong, C.M. Peters, “ Mitogenic signaling by ATP/P2Y purinergic receptors in astrocytes: involvement of a calcium-independent protein kinase C, extracellular signal-regulated protein kinase pathway distinct from the phosphatidylinositol-specific phospholipase C/calcium pathway” JNeuroscience, Vol. 19, No. 11, 1999, pp. 4211-4220.
F. Cavaliere, V. Nestola, S. Amadio, N. D'Ambrosi, DF. Angelini, G. Sancesario, G. Bernardi, C. Volonté, “The metabotropic P2Y4 receptor participates in the commitment to differentiation and cell death of human neuroblastoma SH-SY5Y cells”Neurobiol Dis., Vol. 18, No. 1,2005,pp. 100-109.
J. Milosevic, A. Brandt, U. Roemuss, A. Arnold, F. Wegner, S.C. Schwarz, A. Storch, E. Zimmermann, J. Schwarz, “Uracil nucleotides stimulate human neural precursor cell proliferation and dopaminergic differentiation: involvement of MEK/ERK signaling” J Neurochem, Vol.99, No. 3,2006, pp. 913-923.
S. Yitzhaki, E. Hochhauser, E. Porat, A. Shainberg, “Uridine-5’-triphosphate (UTP) maintains cardiac mitochondrial function following chemical and hypoxic stress” J Mol Cell Cardiol, Vol. 43, No. 5,2007, pp. 653-662.
H.Y. Cheng, M.T. Hsieh, C.R. Wu, FH. Tsai, T.C. Lu, C.C. Hsieh, W.C. Li, Y.T. Lin, W.H. Peng, “Schizandrin protects primary cultures of rat cortical cells from glutamate-induced excitotoxicity” Journal of Pharmacological Science, Vol. 107, No. 1,2008, pp. 21-31.