Protective effects of glucosamine and its acetylated derivative on serum/glucose deprivation-induced PC12 cells death: Role of reactive oxygen species

Seyed Hadi Mousavi, Elham Bakhtiari, Azar Hosseini, Khadijeh Jamialahmadi

Abstract


Finding products with antiapoptotic activities has been one of the approaches for the treatment of neurodegenerative disorders. Serum/glucose deprivation (SGD) has been used as a model for the investigation of the molecular mechanisms of neuronal ischemia. Recent studies indicated that glucosamine (GlcN) and N-acetyl glucosamine (GlcNAc) have many pharmacological effects including antioxidant activities. The present study aimed to investigate the protective effects of GlcN and GlcNAc against SGD-induced PC12 cells injury. The PC12 cells were pretreated with GlcN and GlcNAc for 2 h, and then exposed to SGD for 6, 12 and 24 h. Cell viability was evaluated by MTT assay. The level of intracellular reactive oxygen species (ROS) was determined by flow cytometry using 2',7'- dichlorofluorescin diacetate (DCFH-DA) as a probe. SGD condition caused a significant reduction in cell survival after 6, 12, and 24 h (P < 0.001). Pretreatment with GlcN and GlcNAc (0.6-20 mM) increased cell viability following SGD insult. A significant increase in cell apoptosis was observed in cells under SGD condition after 12 h (P < 0.001). Pretreatment with GlcN and GlcNAc (5-20 mM) decreased apoptosis following SGD condition after 12 h. SGD resulted in a significant increase in intracellular ROS production after 12 h. Pretreatment with both amino sugars at concentrations of 10 to 20 mM could reverse the ROS increment. Results indicated that GlcN and GlcNAc had a cytoprotective property against SGD-induced cell death via anti-apoptosis and antioxidant activities, suggesting that these aminosugers have the potential to be used as novel therapeutic agents for neurodegenerative disorders.


Keywords


Glucosamine; N-acetyl glucosamine; PC12; Serum/glucose deprivation; Apoptosis

Full Text:

PDF

References


Amantea D, Marrone MC, Nistico R, Federici M, Bagetta G, Bernardi G, et al. Oxidative stress in stroke pathophysiology validation of hydrogen peroxide metabolism as a pharmacological target to afford neuroprotection. Int Rev Neurobiol. 2005;85:363-374.

Moley KH, Mueckler MM. Glucose transport and apoptosis. Apoptosis. 2000;5(2): 99–105.

Chu LF, Wang WT, Ghanta VK, Lin CH, Chiang YY, Hsueh CM. Ischemic brain cell-derived conditioned medium protects astrocytes against ischemia through GDNF/ERK/NF-kB signaling pathway. Brain Res. 2008;1239: 24-35.

Hillion JA, Takahashi K, Maric D, Ruetzler C, Barker JL, Hallenbeck JM. Development of an ischemic tolerance model in a PC12 cell line. J Cereb Blood Flow Metab. 2005 25(2):154-162.

Reimann-Philipp U, Ovase R, Weigel PH, Grammas P. Mechanisms of cell death in primary cortical neurons and PC12 cells. J Neurosci Res. 2001;64(6):654-660.

Woronowicz A, Amith SR, Davis VW, Jayanth P, De Vusser K, Laroy W, et al. Trypanosome trans-sialidase mediates neuroprotection against oxidative stress, serum/glucose deprivation, and hypoxia-induced neurite retraction in Trk-expressing PC12 cells. Glycobiol. 2007;17(7):725-734.

Bissett DL, Robinson LR, Raleigh PS, Miyamoto K, Hakozaki T, Li J, et al. Reduction in the appearance of facial hyperpigmentation by topical N-Acetyl glucosamine. J Cosmet Dermatol. 2007;6:20–26.

Hwang YP, Kim HG, Han EH, Choi JH, Park BH, Jung KH, et al. N-acetylglucosamine suppress collagenases activation in ultraviolet B-irradiated human dermal fibroblasts: Involvement of calcium ions and mitogen-activated protein kinases. J Dermatol Sci. 2011;63:93-103.

Ma L, Rudert WA, Harnaha J, Wright M, Machen J, Lakomy R, et al. Immunosuppressive effects of glucosamine. J Biol Chem 2002;277(42):39343-39349.

Yan Y, Wanshun L, Baoqin H, Changhong W, Chenwei F, Bing L et al. The antioxidative and immunostimulating properties of D-glucosamine. Int immunopharmacol. 2007;7:29–35.

Salazar J, Bello L, Chavez M, Anez R, Rojas J, Bermudez V. Glucosamine for osteoarthritis: biological effects, clinical efficacy, and safety on glucose metabolism. Arthritis. 2014;2014:432463.

Xing R, Liu S, Guo Z, Yu H, Li C, Ji X, et al. The antioxidant activity of glucosamine hydrochloride in vitro. Bioorg Med Chem. 2006;14(6):1706-1709.

Mendis E, Kim MM, Rajapakse N, Kim SK. Sulfated glucosamine inhibits oxidation of biomolecules in cells via a mechanism involving intracellular free radical scavenging.Eur J .Pharmacol. 2008;579(1-3):74-85.

Bissett DL. Glucosamine: an ingredient with skin and other benefits. J Cosmet Dermatol. 2006;5: 309–315.

Forchhammer L, Thorn M, Met O, Gad M, Weidner M, Claesson MH. Immunobiological effects of glucosamine in vitro. Scand J Immunol. 2003;58(4):404-411.

Chesnokov V, Gong B, Sun C, Itakura K. Anti-cancer activity of glucosamine through inhibition of N-linked glycosylation. Cancer Cell Int. 2014;14:45.

Champattanachai V, Marchase RB, Chatham JC. Glucosamine protects neonatal cardiomyocytes from ischemia-reperfusion injury via increased protein O-GlcNAc and increased mitochondrial Bcl-2. Am J Physiol Cell Physiol. 2008;294(6):C1509-C20.

Jamialahmadi K, Soltani F, Nabavi Fard M, Behravan J, Mosaffa F. Assessment of protective effects of glucosamine and N-acetyl glucosamine against DNA damage induced by hydrogen peroxide in human lymphocytes. Drug Chem Toxicol. 2014;37(4):427-432.

Jamialahmadi K, Sadeghnia HR, Mohammadi G, Kazemabad AM, Hosseini M. Glucosamine alleviates scopolamine induced spatial learning and memory deficits in rats. Pathophysiology. 2013; 20(4):263-267.

Salvatore S, Heuschkel R, Tomlin S, Davies S, Edwards S, Walker‐Smith JA, et al. A pilot study of N‐acetyl glucosamine, a nutritional substrate for glycosaminoglycan synthesis, in paediatric chronic inflammatory bowel disease. Aliment Pharmacol Ther. 2000;14(12):1567-1579.

Selvan T, Rajiah K, Nainar MS, Mathew EM. A clinical study on glucosamine sulfate versus combination of glucosamine sulfate and NSAIDs in mild to moderate knee osteoarthritis. Sci World J. 2012;2012:902676.

Chen JK, Shen CR, Liu CL. N-acetylglucosamine: production and applications. Mar Drugs. 2010;8:2493-251.

Shikhman AR, Amiel D, D'Lima D, Hwang SB, Hu C, Xu A. Chondroprotective activity of N -acetylglucosamine in rabbits with experimental osteoarthritis. Ann Rheum Dis. 2005;64:89-94.

Sayo T, Sakai S, Inoue S. Synergestic effect of N-acetylglucosamine and retinoids on hyaluronan production in human keratinocytes. Skin Pharmacol Physiol. 2004;17:77–83.

Kamel M, Alnahdi M. Inhibition of superoxide anion release from human polymorphonuclear leukocytes by N-acetylgalactosamine and N-acetylglucosamine. Clin Rheumatol. 1992;11: 254–260.

Malaekeh‐Nikouei B, Golmohammadzadeh S, Salmani‐Chamanabad S, Mosallaei N, Jamialahmadi K. Preparation, characterization, and moisturizing effect of liposomes containing glucosamine and N‐acetyl glucosamine. J Cosmet Dermatol. 2013;12(2):96-102.

Choi H, Chung M, Kweon Park J, Park Y. Neuroprotective effects of N-acetylglucosamine against hydrogen peroxide-induced apoptosis in human neuronal SK-N-SH cells by inhibiting the activation of caspase-3, PARP, and p38. Food Sci Biotechnol. 2013;22(3):853-858.

Azam MS, Kim EJ, Yang HS, Kim JK. High antioxidant and DNA protection activities of N-acetylglucosamine (GlcNAc) and chitobiose produced by exolytic chitinase from Bacillus cereus EW5. Springerplus. 2014;3:354.

Mousavi SH, Tayarani-Najaran Z, Asghari M, Sadeghnia HR. Protective effect of Nigella sativa extract and thymoquinone on serum/glucose deprivation-induced PC12 cells death. Cell Mol Neurobiol. 2010;30(4):591-598.

Sarvmeili N, Jafarian-Dehkordi A, Zolfaghari B. Cytotoxic effects of Pinus eldarica essential oil and extracts on HeLa and MCF-7 cell lines. Res Pharm Sci. 2016;11(6):476-483.

Rosamond W, Flegal K, Friday G, Furie K, Go A, Greenlund K, et al. Heart disease and stroke statistics 2007 update: a report from the American Heart Association Statistics Committee and Stroke Statistics Subcommittee. Circulation. 2007;115(5):e69-171.

Arabsolghar R, Saberzadeh J, Khodaei F, Abbasi Borojeni R, Khorsand M, Rashedinia M. The protective effect of sodium benzoate on aluminum toxicity in PC12 cell line. Res Pharm Sci. 2017; 12(5): 391–400.

Forouzanfar F, Afkhami Goli A, Asadpour E, Ghorbani A, Sadeghnia HR. Protective Effect of Punica granatum L. against serum/glucose deprivation-induced PC12 cells injury. Evid Based Complement Alternat Med. 2013;2013:716730.

Bakhtiari E, Hosseini A, Mousavi SH. Protective effect of Hibiscus sabdariffa against serum/glucose deprivation-induced PC12 cells injury. Avicenna J Phytomed. 2015;5(3):231-237.

Lorenz L, Dang J, Misiak M, Tameh Abolfazl A, Beyer C, Kipp M. Combined 17beta-oestradiol and progesterone treatment prevents neuronal cell injury in cortical but not midbrain neurons or neuroblastoma cells. J Neuroendocrinol. 2009;21(10):841-849.

Jamialahmadi K, Arasteh O, Riahi MM, Mehri S, Riahi-Zanjani B, Karimi G. Protective Effects of glucosamine hydrochloride against free radical-induced erythrocytes damage. Environ Toxicol Pharmacol. 2014;38:212-219.


Refbacks

  • There are currently no refbacks.


Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

Creative Commons LicenseThis work is licensed under a Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International License which allows users to read, copy, distribute and make derivative works for non-commercial purposes from the material, as long as the author of the original work is cited properly.