Neuroprotective effects of Salvia aristata Aucher ex Benth. on hydrogen peroxide induced apoptosis in SH-SY5Y neuroblastoma cells

Document Type: Original paper

Authors

Department of Biology, Medicinal Plants and Drug Research Institute, Shahid Beheshti University G. C., Tehran, Iran.

Abstract

Background and objectives: Oxidative stress is implicated in the neuronal damage associated with Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, amyotropic lateral sclerosis and cerebral ischemic stroke. The present work was designed to establish the neuroprotective effects of Salvia aristata extract on H2O2-induced apoptosis in human dopaminergic SH-SY5Y cells. Methods: The total phenol and flavonoids contents of the plant extracts were quantified by colorimetric methods. The antioxidant activity was assessed using DPPH free radicals scavenging activity assay, and the neuroprotective effect on H2O2-induced oxidative stress was also investigated using human dopaminergic SH-SY5Y cells by MTT assay and western blotting techniques. Results: The highest scavenging activity was found for methanol extract of S. aristata roots (85.28 ± 2.61 μg/mL), with the highest total phenolic and flavonoids content (90.28 mg total phenols as gallic acid and 250.12 mg total flavonoids as rutin, respectively). Our results also, showed that H2O2-induced cytotoxicity in SH-SY5Y cells was suppressed by treatment with S. aristata. Moreover, S. aristata root extract was effective in attenuating the disruption of mitochondrial membrane potential and apoptotic cell death has induced by H2O2S. aristata suppressed the down-regulation of Bcl-2, upregulation of Bax, and the release of mitochondrial cytochrome c to cytosol. In addition, S. aristata attenuated caspase-3, and -9 activation, and eventually protected the cells against H2O2-induced apoptosis. Conclusion: Theresults of the present study suggest that treatment of SH-SY5Y cells with S. aristata could block H2O2-induced apoptosis by regulating Bcl-2 family members and by suppressing caspase cascade activation.

Keywords


[1] Uttara B, Singh AV, Zamboni P, Mahajan RT. Oxidative stress and neurodegenerative diseases: a review of upstream and downstream antioxidant therapeutic options. Curr Neuropharmacol.2009; 7(1): 65-74.

[2] Fordel E, Thijs L, Martinet W, Lenjou M, Laufs T, Van Bockstaele D, Moens L, Dewilde S. Neuroglobin and cytoglobin overexpression protects human SH-SY5Y neuroblastoma cells against oxidative stress-induced cell death. Neurosci Lett. 2006; 410(2): 146-151.

[3] Kwon SH, Kim JA, Hong SI, Jung YH, Kim HC, Lee SY, Jang CG. Loganin protects against hydrogen peroxide-induced apoptosis by inhibiting phosphorylation of JNK, p38, and ERK 1/2 MAPKs in SH-SY5Y cells. Neurochem Int. 2011; 58(4): 533-541.

[4] King KL, Cidlowski JA. Cell cycle and apoptosis: common pathways to life and death. J Cell Biochem. 1995; 58(2): 175-180.

[5] Healy D. Meyler's side effects of drugs, 12th edition. Hum Psychopharm Clin. 1995; 10(6): 494.

[6] Shankar S, Kumar D, Srivastava RK. Epigenetic modifications by dietary phytochemicals: implications for personalized nutrition. Pharmacol Therapeut. 2013; 138(1): 1-17.

[7] Banthorpe DV, Bilyard HJ, Brown GD. Enol esters of caffeic acid in several genera of the Labiatae. Phytochemistry. 1989; 28(8): 2109-2113.

[8] Ulubelen A, Topcu G. Abietane diterpenoids from Salvia pomifera. Phytochemistry. 1992; 31(11): 3949-3951.

[9] Bozin B, Mlmica-Dukic N, Samojlik I, Jovin E. Antimicrobial and antioxidant properties of rosemary and sage (Rosmarinus officinalis L. and Salvia officinalis L., lamiaceae) essential oils. J Agr Food Chem. 2007; 55(19): 7879-7885.

[10] Slinkard K, Singleton VL. Total phenol analysis: automation and comparison with manual methods. J Enol Vitic. 1977; 28(1): 49-55.

[11] Maksimovic Z, Malencic D, Kovacevic N. Polyphenol contents and antioxidant activity of Maydis stigma extracts. Bioresource Technol. 2005; 96(8): 873-877.

[12] Blois MS. Antioxidant determinations by the use of a stable free radical. Nature. 1958; 181: 1199-1200.

[13] Rafatian G, Khodagholi F, Farimani MM, Abraki SB, Gardaneh M. Increase of autophagy and attenuation of apoptosis by Salvigenin promote survival of SH-SY5Y cells following treatment with H2O2. Mol Cell Biochem. 2012;371(1-2): 9-22.

[14] Rice-Evans C, Miller N, Paganga G. Antioxidant properties of phenolic compounds. Trends Plant Sci. 1997; 2(4): 152-159.

[15] Pietta PG. Flavonoids as antioxidants. J Nat Prod. 2000; 63(7): 1035-1042.

[16] Liyana-Pathirana C, Dexter J, Shahidi F. Antioxidant properties of wheat as affected by pearling. J Agr Food Chem. 2006; 54(17): 6177-6184.

[17] Biedler JL, Helson L, Spengler BA. Morphology and growth, tumorigenicity, and cytogenetics of human neuroblastoma cells in continuous cultureCancer Res. 1973; 33(11): 2643-2652. 

[18] Chow JM, Shen SC, Huan SK, Lin HY, Chen YC. Quercetin, but not rutin and quercitrin, prevention of H2O2-induced apoptosis via anti-oxidant activity and heme oxygenase 1 gene expression in macrophages. Cancer Res. 2005; 69(12):1839-1851.

[19] Garcia-Alonso M, Jacobs E, Raybould A, Nickson TE, Sowig P, Willekens H, Van der Kouwe P, Layton R, Amijee F, Fuentes AM, Tencalla F. A tiered system for assessing the risk of genetically modified plants to non-target organisms. Environ Biosafety Res. 2006; 5(2): 57-65.

[20] Vento R, Giuliano M, Lauricella M, Carabillò M, Di Liberto D, Tesoriere G. Induction of programmed cell death in human retinoblastoma Y79 cells by C2-ceramide. Mol Cell Biochem. 1998; 185(1-2): 7-15.

[21] Shimohama S, Fujimoto S, Sumida Y, Tanino H. Differential expression of rat brain bcl-2 family proteins in development and aging. Biochem Bioph Res Co. 1998; 252(1): 92-96.

[22] Kitamura Y, Shimohama S, Kamoshima W, Ota T, Matsuoka Y, Nomura Y, Smith MA, Perry G, Whitehouse PJ, Taniguchi T. Alteration of proteins regulating apoptosis, Bcl-2, Bcl-x, Bax, Bak, Bad, ICH-1 and CPP32, in Alzheimer’s disease. Brain Res. 1998; 780: 260-269.

[23] Gogvadze V, Orrenius S, Zhivotovsky B. Multiple pathways of cytochrome c release from mitochondria in apoptosis. Biochim Biophys Acta. 2006; 1757(5-6): 639-647.

[24] Kagan VE, Bayir HA, Belikova NA, Kapralov O, Tyurina YY, Tyurin VA, Jiang J, Stoyanovsky DA, Wipf P, Kochanek PM, Greenberger JS, Pitt B, Shvedova AA, Borisenko G. Cytochrome c/cardiolipin relations in mitochondria: a kiss of death. Free Radic Biol Med. 2009; 46(11): 1439-1453.

[25] Brown GC, Borutaite V. Regulation of apoptosis by the redox state of cytochrome cBiochim Biophys Acta. 2008; 1777(7-8): 877-881.

[26] Jiang XJ, Wang XD. Cytochrome C-mediated apoptosis. Annu Rev Biochem. 2004; 73: 87-106.

[27] Shimizu S, Narita M, Tsujimoto Y. Bcl-2 family proteins regulate the release of apoptogenic cytochrome c by the mitochondrial channel VDAC. Nature.1999; 399(6735): 483-487.