MMP9 Gene Expression Variation by Ingesting Tart Cherry and P-Coumaric Acid During Remyelination in the Cuprizone Mouse Model

  • Newshan Behrangi Faculty of Basic Sciences and Advanced Technologies in Biology, University of Sciences and Culture, Tehran, Iran.
  • Nabiyollah Namvar Pasteur Institute, Production and Research Complex, Karaj, Iran.
  • Mitra Ataei National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
  • Sakineh Dizaji Pasteur Institute, Production and Research Complex, Karaj, Iran.
  • Golshid Javdani Pasteur Institute, Production and Research Complex, Karaj, Iran.
  • Mahommad Hossein Sanati National Institute of Genetic Engineering and Biotechnology, Tehran, Iran.
Keywords: Multiple sclerosis, MMP9, Cuprizone, P-coumaric acid, Tart cherry


Matrix metalloproteinase-9 (GELB) as a member of gelatinases plays key role in the destruction of blood-brain barrier (BBB), T cells migration into the CNS, and demyelination induction. Considering remyelination induction in response to tart cherry extract and pure p-coumaric acid ingestion via tracking MMP9 gene expression in the cuprizone mouse model. Firstly, predicting the chemical interaction between p-coumaric acid and MMP9 protein was conducted through PASS and Swiss dock web services. Next, the content of p-coumaric acid in the tart cherry extract was analyzed by HPLC. Later, mice (male, female) were categorized into two groups: standard, cuprizone. After the demyelination period, mice classified into four groups: standard, natural chow, tart cherry extract, p-coumaric acid. Finally, brains were extracted from the skull, and MMP9 gene expression was evaluated by real time RT-PCR. Bioinformatics analysis displayed p-coumaric acid has potent inhibitory effect on MMP9 gene expression (Pa=0.818) with estimated ΔG (kcal/mol) -8.10. In addition, during the demyelination period, MMP9 expression was increased significantly in the male group that is related to myelin destruction. However, MMP9 was declined throughout remyelination in both male and female. It’s remarkable that pure p-coumaric acid and tart cherry extract ingestion could decrease the gene expression ratio more than natural chow. According to the results, it’s deduced the male mouse is more appropriate gender for demyelination induction via cuprizone. In addition, tart cherry extract and pure p-coumaric acid ingestion could decrease MMP9 gene expression level considerably during remyelination.


Van der Valk, P. and C. De Groot, Staging of multiple sclerosis (MS) lesions: pathology of the time frame of MS. Neuropathology and applied neurobiology, 2000. 26(1): p. 2-10.

Kipp, M., et al., The cuprizone animal model: new insights into an old story. Acta neuropathologica, 2009. 118(6):723-736.

Borkakoti, N., Matrix metalloproteases: variations on a theme. Progress in biophysics and molecular biology, 1998. 70(1):73-94.

Lindberg, R.L., et al., The expression profile of matrix metalloproteinases (MMPs) and their inhibitors (TIMPs) in lesions and normal appearing white matter of multiple sclerosis. Brain, 2001. 124(9):1743-1753.

Rosenberg, G.A., Matrix metalloproteinases and neuroinflammation in multiple sclerosis. The Neuroscientist, 2002. 8(6):586-595.

Leppert, D., et al., Matrix metalloproteinase-9 (gelatinase B) is selectively elevated in CSF during relapses and stable phases of multiple sclerosis. Brain, 1998. 121(12):2327-2334.

Leco, K.J., et al., Matrix metalloproteinase‐9 maps to the distal end of chromosome 2 in the mouse. Developmental genetics, 1997. 21(1):55-60.

Ortega, N., et al., Galectin-3 is a downstream regulator of matrix metalloproteinase-9 function during endochondral bone formation. Molecular biology of the cell, 2005. 16(6):3028-3039.

Nagase, H. and J.F. Woessner, Matrix metalloproteinases. Journal of Biological Chemistry, 1999. 274(31):21491-21494.

Baeten, K.M. and K. Akassoglou, Extracellular matrix and matrix receptors in blood–brain barrier formation and stroke. Developmental neurobiology, 2011. 71(11):1018-1039.

Ramos-DeSimone, N., et al., Activation of matrix metalloproteinase-9 (MMP-9) via a converging plasmin/stromelysin-1 cascade enhances tumor cell invasion. Journal of Biological Chemistry, 1999. 274(19):13066-13076.

Cossins, J.A., et al., Enhanced expression of MMP-7 and MMP-9 in demyelinating multiple sclerosis lesions. Acta neuropathologica, 1997. 94(6):590-598.

Leppert, D., et al., T cell gelatinases mediate basement membrane transmigration in vitro. The Journal of Immunology, 1995. 154(9):4379-4389.

Lichtinghagen, R., et al., Expression of matrix metalloproteinase-9 and its inhibitors in mononuclear blood cells of patients with multiple sclerosis. Journal of neuroimmunology, 1999. 99(1):19-26.

Gijbels, K., et al., Gelatinase in the cerebrospinal fluid of patients with multiple sclerosis and other inflammatory neurological disorders. Journal of neuroimmunology, 1992. 41(1):29-34.

Boz, C., et al., Matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of matrix metalloproteinase (TIMP-1) in patients with relapsing–remitting multiple sclerosis treated with interferon beta. Clinical neurology and neurosurgery, 2006. 108(2):124-128.

Ferretti, G., et al., Cherry antioxidants: from farm to table. Molecules, 2010. 15(10):6993-7005.

Mulabagal, V., et al., Anthocyanin content, lipid peroxidation and cyclooxygenase enzyme inhibitory activities of sweet and sour cherries. Journal of agricultural and food chemistry, 2009. 57(4):1239-1246.

Wang, H., et al., Novel Antioxidant Compounds from Tart Cherries (Prunus c erasus). Journal of Natural Products, 1999. 62(1):86-88.

Haidari, F., et al., Inhibitory Effects of Tart Cherry (Prunus cerasus) Juice on Xanthine Oxidoreductase Activity and its Hypouricemic and Antioxidant Effects on Rats. Malaysian journal of nutrition, 2009. 15(1):53-64.

Šarić, A., et al., Improved antioxidant and anti-inflammatory potential in mice consuming sour cherry juice (Prunus Cerasus cv. Maraska). Plant foods for human nutrition, 2009. 64(4):231-237.

Tall, J.M. and S.N. Raja, Dietary constituents as novel therapies for pain. The Clinical journal of pain, 2004. 20(1):19-26.

Kuehl, K.S., et al., Research article Efficacy of tart cherry juice in reducing muscle pain during running: a randomized controlled trial. Journal of the International Society of Sports Nutrition, 2010. 7:17.

Kim, D.-O., et al., Sweet and sour cherry phenolics and their protective effects on neuronal cells. Journal of Agricultural and Food Chemistry, 2005. 53(26):9921-9927.

Heo, H.J., et al., Potent inhibitory effect of flavonoids in Scutellaria baicalensis on amyloid β protein-induced neurotoxicity. Journal of Agricultural and Food Chemistry, 2004. 52(13):4128-4132.

Mehrdad, H., Evaluating new targets of natural anticancer molecules through bioinformatics tools. Journal of Proteomics & Bioinformatics, 2012. 5(2) : 050-053

Behrangi, N., et al., Comparison among Cornelian Cherry and Prunus cerasus According to Phenolic Content and Antioxidant Capacity by Three Various Methods of Extraction. Food and Nutrition Sciences, 2015. 6(12):1166-1173.

Gudi, V., et al., Glial response during cuprizone-induced de-and remyelination in the CNS: lessons learned. Frontiers in cellular neuroscience, 2014. 8:73.

Herring, N.R. and C. Konradi, Myelin, copper, and the cuprizone model of schizophrenia. Frontiers in bioscience (Scholar edition), 2011. 3:23.

Gudi, V., et al., Regional differences between grey and white matter in cuprizone induced demyelination. Brain research, 2009. 1283:127-138.

Antonio, M., et al., A rational approach on the use of sex steroids in multiple sclerosis. Recent patents on CNS drug discovery, 2008. 3(1): 34-39.

Duquette, P. and M. Girard, Hormonal factors in susceptibility to multiple sclerosis. Current Opinion in Neurology, 1993. 6(2):195-201.

Harsan, L.-A., et al., Recovery from chronic demyelination by thyroid hormone therapy: myelinogenesis induction and assessment by diffusion tensor magnetic resonance imaging. The Journal of neuroscience, 2008. 28(52):14189-14201.

Taylor, L.C., et al., Cuprizone induces similar demyelination in male and female C57BL/6 mice and results in disruption of the estrous cycle. Journal of neuroscience research, 2010. 88(2):391-402.

Praet, J., et al., Cellular and molecular neuropathology of the cuprizone mouse model: clinical relevance for multiple sclerosis. Neuroscience & Biobehavioral Reviews, 2014. 47:485-505.

Romanic, A.M., et al., Matrix metalloproteinase expression increases after cerebral focal ischemia in rats inhibition of matrix metalloproteinase-9 reduces infarct size. Stroke, 1998. 29(5):1020-1030.

Rosenberg, G.A., et al., Injury-induced 92-kilodalton gelatinase and urokinase expression in rat brain. Laboratory investigation; a journal of technical methods and pathology, 1994. 71(3):417-422.

Chandler, S., et al., Matrix metalloproteinases degrade myelin basic protein. Neuroscience letters, 1995. 201(3):223-226.

Gijbels, K., et al., Gelatinase B is present in the cerebrospinal fluid during experimental autoimmune encephalomyelitis and cleaves myelin basic protein. Journal of neuroscience research, 1993. 36(4):432-440.

Proost, P., J. Vandamme, and G. Opdenakker, Leukocyte gelatinase B cleavage releases encephalitogens from human myelin basic protein. Biochemical and biophysical research communications, 1993. 192(3):1175-1181.

Larsen, P.H., et al., Myelin formation during development of the CNS is delayed in matrix metalloproteinase-9 and-12 null mice. The Journal of neuroscience, 2006. 26(8):2207-2214.

Larsen, P.H. and V.W. Yong, The expression of matrix metalloproteinase-12 by oligodendrocytes regulates their maturation and morphological differentiation. The Journal of neuroscience, 2004. 24(35):7597-7603.

Larsen, P.H., et al., Matrix metalloproteinase-9 facilitates remyelination in part by processing the inhibitory NG2 proteoglycan. The Journal of neuroscience, 2003. 23(35):11127-11135.

de Santana Nunes, A.K., et al., Phosphodiesterase-5 inhibition promotes remyelination by MCP-1/CCR-2 and MMP-9 regulation in a cuprizone-induced demyelination model. Experimental neurology, 2016. 275:143-153.

Saja, K., et al., Anti-inflammatory effect of curcumin involves downregulation of MMP-9 in blood mononuclear cells. International immunopharmacology, 2007. 7(13):1659-1667.

Pragasam, S.J., V. Venkatesan, and M. Rasool, Immunomodulatory and anti-inflammatory effect of p-coumaric acid, a common dietary polyphenol on experimental inflammation in rats. Inflammation, 2013. 36(1):169-176.

Jakobek, L., et al., Flavonols, phenolic acids and antioxidant activity of some red fruits. Deutsche Lebensmittel-Rundschau, 2007. 103(8):369-378.

Rocha, L.D., M.C. Monteiro, and A.J. Teodoro, Anticancer properties of hydroxycinnamic acids-A Review. Cancer and clinical oncology, 2012. 1(2):109.

Vauzour, D., G. Corona, and J.P. Spencer, Caffeic acid, tyrosol and p-coumaric acid are potent inhibitors of 5-S-cysteinyl-dopamine induced neurotoxicity. Archives of Biochemistry and Biophysics, 2010. 501(1):106-111.

Lawrence, T., The nuclear factor NF-κB pathway in inflammation. Cold Spring Harbor perspectives in biology, 2009. 1(6):1651.

Chou, Y.-C., et al., Nuclear-targeted inhibition of NF-κB on MMP-9 production by N-2-(4-bromophenyl) ethyl caffeamide in human monocytic cells. Chemico-biological interactions, 2010. 184(3):403-412.

Ma, Q. and K. Kinneer, Chemoprotection by Phenolic Antioxidants INHIBITION OF TUMOR NECROSIS FACTOR α INDUCTION IN MACROPHAGES. Journal of Biological Chemistry, 2002. 277(4):2477-2484.

Yoon, J.-H., et al., p-Coumaric Acid and Ursolic Acid from Corni fructus Attenuated β-Amyloid25–35-Induced Toxicity through Regulation of the NF-κB Signaling Pathway in PC12 Cells. Journal of agricultural and food chemistry, 2014. 62(21):4911-4916.

Lua, F.C., et al., Attenuation of iNOS and COX2 of blueberry polyphenols is mediated through the supression of NF-kB activation. Journal of function foods, 2009:274-283.

Wang, L., et al., Natural products as a gold mine for selective matrix metalloproteinases inhibitors. Bioorganic & medicinal chemistry, 2012. 20(13):4164-4171.

How to Cite
Behrangi N, Namvar N, Ataei M, Dizaji S, Javdani G, Sanati MH. MMP9 Gene Expression Variation by Ingesting Tart Cherry and P-Coumaric Acid During Remyelination in the Cuprizone Mouse Model. Acta Med Iran. 55(9):539-549.