The mechanism of neuroprotective effect of Viola odorata against serum/glucose deprivation-induced PC12 cell death

Document Type : Short communication

Authors

1 Biotechnology Research Center, Pharmaceutical Technology Institute, Mashhad University of Medical Sciences, Mashhad, Iran.

2 Molecular and Cell Biology Research Center, Faculty of Medicine, Mazandaran University of Medical Sciences, Sari, Iran.

3 Department of Cellular and Molecular Biology, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran.

4 Department of Biology, Faculty of Sciences, Mashhad Branch, Islamic Azad University, Mashhad, Iran.

5 Department of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran

6 Department of Traditional Pharmacy, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.

Abstract

Objective: Oxidative stress is associated with the pathogenesis of brain ischemia and other neurodegenerative disorders. Previous researches have shown the antioxidant activity of Viola odorata L. In this project, we studied neuro-protective and reactive oxygen species (ROS) scavenging activities of methanol  (MeOH) extract and other fractions isolated from V. odorata in PC12 cell line in serum/glucose deprivation  (SGD) condition.
Materials and Methods: The PC12 neuronal cells were pretreated for 6 hr with MeOH extract and fractions of V. odorata (1 to 25 μg/ml) followed by 24 hr incubation under SGD condition. Cell viability was measured by Alamar Blue® assay. The level of ROS was calculated using DCFH-DA. Also, Bax/Bcl-2 protein ratio was analyzed by western blot assay.
Results: SGD condition significantly decreased cells viability (p<0.001). Pretreatment with EtOAc (12.5 and 25 µg/ml), BuOH (12, 25, 50 µg/ml) and CH2Cl2 (1.5 µg/ml) fractions of V. odorata reduced SGD-induced cytotoxicity. MeOH extract could not increase the viability significantly. All four semi polar fractions (EtOAc, BuOH, CH2Cl2 and MeOH) decreased SGD-induced ROS production and changed Bax/Bcl-2 ratio.
Conclusion: V. odorata showed promising effects against SGD condition; further mechanistic and clinical studies are warranted before application of V. odorata as a neuro-protective agent.

Keywords

Main Subjects


Alinejad B, Ghorbani A, Sadeghnia HR. 2013. Effects of combinations of curcumin, linalool, rutin, safranal, and thymoquinone on glucose/serum deprivation-induced cell death. Avicenna J Phytomed, 3: 321-328.
Anoopkumar-Dukie S, Carey J, Conere T, O'sullivan E, Van pelt F, Allshire A. 2005. Resazurin assay of radiation response in cultured cells. Br J Radiol, 78: 945-947.
Bors W, Michel C, Saran M. 1984. Inhibition of the bleaching of the carotenoid crocin a rapid test for quantifying antioxidant activity. Biochim Biophys Acta, 796: 312-319.
Chandra D, Kohli G, Prasad K, Bisht G, Punetha VD, Khetwal K, Devrani MK, Pandey H. 2015. Phytochemical and ethnomedicinal uses of family Violaceae. Curr Res Chem, 7:44-52.
Collins AR, Rašlová KN, Somorovská M, Petrovská H, Ondrušová A, Vohnout B, Fábry R, Dušinská M. 1998. DNA damage in diabetes: correlation with a clinical marker. Free Radic Biol Med, 25: 373-377.
Ebrahimzadeh MA, nabavi SM, nabavi SF, Bahramian F, Bekhradnia AR. 2010. Antioxidant and free radical scavenging activity of H. officinalis L. var. angustifolius, V. odorata, B. hyrcana and C. speciosum. Pak J Pharm Sci, 23:29-34.
Ekshyyan O, Aw TY. 2004. Apoptosis in acute and chronic neurological disorders. Front Biosci, 9: 1567-1576.
Galato D, Ckless K, Susin MF, Giacomelli C, Ribeiro-DO-Valle RM, Spinelli A. 2001. Antioxidant capacity of phenolic and related compounds: correlation among electrochemical, visible spectroscopy methods and structure–antioxidant activity. Redox Report, 6: 243-250.
Halliwell B. 1992. Reactive oxygen species and the central nervous system. J Neurochem, 59: 1609-1623.
Hillion JA, Takahashi K, Maric D, Ruetzler C, Barker JL, Hallenbeck JM. 2005. Development of an ischemic tolerance model in a PC12 cell line. J Cereb Blood Flow Metab, 25: 154-162.
Hockenbery DM, Oltvai ZN, Yin, X-M, Milliman CL, Korsmeyer SJ. 1993. Bcl-2 functions in an antioxidant pathway to prevent apoptosis. Cell, 75:241-251.
 
Hosseinzadeh H, Shamsaie F, Mehri S. 2009. Antioxidant activity of aqueous and ethanolic extracts of Crocus sativus L. stigma and its bioactive constituents, crocin and safranal. Pharmacogn Mag, 5: 419-424.
Javadi B, Iranshahy M, Emami SA. 2015. Anticancer plants in Islamic traditional medicine. Complementary Therapies for the Body, Mind and Soul. IntechOpen.
Koochek M, Pipelzadeh M, Mardani H. 2003. The effectiveness of Viola odorata in the prevention and treatment of formalin-induced lung damage in the rat. J Herbs Spices Med Plants, 10:95-103.
Mórocz M, Kálmán J, Juhász A, Sinkó I, Mcglynn AP, Downes, CS, Janka Z, Raskó I. 2002. Elevated levels of oxidative DNA damage in lymphocytes from patients with Alzheimer’s disease. Neurobiol Aging, 23: 47-53.
Mousavi SH, Tayarani N, Parsaee H. 2010. Protective effect of saffron extract and crocin on reactive oxygen species-mediated high glucose-induced toxicity in PC12 cells. Cell Mol Neurobiol, 30:185-191.
Pham-Huy LA, He H, Pham-Huy C. 2008. Free radicals, antioxidants in disease and health.IJBS, 4: 89-96.
Piantadosi CA, Zhang J. 1996. Mitochondrial generation of reactive oxygen species after brain ischemia in the rat. Stroke, 27: 327-332.
Qadir MI, Ali M, Saleem M, Hanif M. 2014. Hepatoprotective activity of aqueous methanolic extract of Viola odorata against paracetamol-induced liver injury in mice. Bangladesh J Pharmacol, 9: 198-202.
Reed JC, Miyashita T, Takayama S, Wang H. G, Sato T, Krajewski S, Aimé‐Sempé C, Bodrug S, Kitada S, Hanada M. 1996. Bcl‐2 family proteins: regulators of cell death involved in the pathogenesis of cancer and resistance to therapy. J Cell Biochem, 60: 23-32.
Sadeghnia HR, Farahm SK, Asadpour E, Rakhsh H, Ghorbani A. 2012. Neuroprotective effect of Lactuca sativa on glucose/serum deprivation-induced cell death. Afr J Pharm Pharmacol, 6: 2464-2471.
Sato T, Hanada M, Bodrug S, Irie S, Iwama N, Boise LH, Thompson CB, Golemis E, Fong L, Wang HG. 1994. Interactions among members of the bcl-2 protein family analyzed with a yeast two-hybrid system. Proc Natacad Sci Usa, 91: 9238-9242.
Sims NR, Muyderman H. 2010. Mitochondria, oxidative metabolism and cell death in stroke. BBA-Molecular Basis of Disease, 1802: 80-91.
Stojković D, Glamočlija J, Ćirić A, Šiljegović, J, Nikolić M, Soković M. 2011. Free radical scavenging activity of Viola odorata water extracts. J Herbs Spices Med Plants, 17: 285-290.
Tayarani-Najaran Z, Sareban M, Gholami A, Emami SA, Mojarrab M. 2013. Cytotoxic and apoptotic effects of different extracts of Artemisia turanica Krasch on K562 and HL-60 cell lines. Sci World J,2013:628073-628079.
Tayarani-Najaran Z, Tayarani-Najaran N, Emami SA. 2014. The history of Islamic medicine at a glance. Polyphenols in Human Health and Disease. Academic Press, 17-27.
Woronowicz A, Amith SR, Davis VW, Jayanth P, DE Vusser K, Laroy W, Contreras R, Meakin SO , Szewczuk MR. 2007. Trypanosome trans-sialidase mediates neuroprotection against oxidative stress, serum/glucose deprivation, and hypoxia-induced neurite retraction in Trk-expressing PC12 cells. Glycobiology, 17:725-734.
Wyllie A. 1972. A basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J cancer, 26: 239-257.