The Effect of Rosmarinus officinalis L Extract on the Inhibition of High Glucose-Induced Neurotoxicity in PC12 Cells: an In Vitro Model of Diabetic Neuropathy

  • Marzieh Rashidipour Razi Herbal Medicines Research Center, Lorestan University of Medical Sciences, Khorramabad, Iran
  • Zahra Hajializadeh Endocrinology and Metabolism Research Center, Institute of Basic and Clinical Physiology Sciences, Kerman University of Medical Sciences, Kerman, Iran
  • Saeed Esmaeili-Mahani Department of Biology, Faculty of Sciences, Shahid Bahonar University of Kerman, Kerman, Iran
  • Ayat Kaeidi * Physiology-pharmacology Research Center, Rafsanjan University of Medical Sciences, Rafsanjan, Iran Department of Physiology and Pharmacology, School of Medicine, Rafsanjan University of Medical Sciences, Rafsanjan, Iran
Keywords: Rosmarinus officinalis, hyperglycemia, cell toxicity, apoptosis, diabetic neuropathy, PC12 cells

Abstract

Background and Aim: Diabetes mellitus and continued hyperglycemic condition prompt serious diabetic complications such as diabetic neuropathy. Numerous studies have shown the involvement of oxidative stress and high glucose-induced cell death in the progress of diabetic neuropathy. Rosmarinus officinalis L has been suggested in the literature as an anti-diabetic herbal medicine that contains potent antioxidant components. Therefore, the neuroprotective effect of Rosmarinus officinalis L (RE) extract was investigated in glucose-induced neurotoxicity via pheochromocytoma )PC12( cells as an appropriate in vitro model of diabetic neuropathy.Materials and Methods: Cell viability was determined using MTT assay. Cleave caspase-3 and Bax: Bcl2 ratio, as biochemical parameters of cellular apoptosis, were measured by western blotting analysis.Results: Our data showed that a 4-fold elevation in the medium glucose significantly reduced cell viability (P < 0.01) and increased caspase-3 activation (P < 0.01) as well as Bax: Bcl2 ratio (P < 0.05) in PC12 cells after 24 h. Incubation of high glucose medium cells with 60μg/ml RE extract decreased high glucose-induced cell toxicity and prevented caspase-3 activation and Bax: Bcl2 ratio.Conclusion: It could be concluded that RE extract is effective in the protection against hyperglycemia-induced cellular toxicity. This could be relevant to the prevention of apoptosis. Moreover, the results suggest that RE has the therapeutic potential to attenuate diabetes complications such as that neuropathy.

References

Vinik AI, Nevoret M-L, Casellini C, Parson H. Diabetic neuropathy. Endocrinol Metab Clin North Am. 2013;42(4):747–87.

Edwards JL, Vincent AM, Cheng HT, Feldman EL. Diabetic neuropathy: mechanisms to management. Pharmacol Ther. 2008;120(1):1–34.

Tomlinson DR, Gardiner NJ. Glucose neurotoxicity. Nat Rev Neurosci. 2008;9(1):36–45.

Russell JW, Golovoy D, Vincent AM, Mahendru PIA, Olzmann JA, Mentzer A, et al. High glucose-induced oxidative stress and mitochondrial dysfunction in neurons. FASEB J. 2002;16(13):1738–48.

Kaeidi A, Esmaeili-Mahani S, Abbasnejad M, Sheibani V, Rasoulian B, Hajializadeh Z, et al. Satureja khuzestanica attenuates apoptosis in hyperglycemic PC12 cells and spinal cord of diabetic rats. J Nat Med. 2013;67(1):61–9.

Kaeidi A, Taati M, Hajializadeh Z, Jahandari F, Rashidipour M. Aqueous extract of Zizyphus jujuba fruit attenuates glucose induced neurotoxicity in an in vitro model of diabetic neuropathy. Iran J Basic Med Sci. 2015;18(3):301.

Kamboj SS, Vasishta RK, Sandhir R. N-acetylcysteine inhibits hyperglycemia-induced oxidative stress and apoptosis markers in diabetic neuropathy. J Neurochem. 2010;112(1):77–91.

Sharifi AM, Eslami H, Larijani B, Davoodi J. Involvement of caspase-8,-9, and-3 in high glucose-induced apoptosis in PC12 cells. Neurosci Lett. 2009;459(2):47–51.

O’Brien NM, Woods JA, Aherne SA, O’Callaghan YC. Cytotoxicity, genotoxicity and oxidative reactions in cell-culture models: modulatory effects of phytochemicals. Portland Press Limited; 2000.

Whaley-Connell A, McCullough PA, Sowers JR. The role of oxidative stress in the metabolic syndrome. Rev Cardiovasc Med. 2010;12(1):21–9.

Ribeiro LR, Salvadori DMF. Dietary components may prevent mutation-related diseases in humans. Mutat Res Mutat Res. 2003;544(2):195–201.

Hajializadeh Z, Nasri S, Kaeidi A, Sheibani V, Rasoulian B, Esmaeili-Mahani S. Inhibitory effect of Thymus caramanicus Jalas on hyperglycemia-induced apoptosis in in vitro and in vivo models of diabetic neuropathic pain. J Ethnopharmacol. 2014;153(3):596–603.

Ablat N, Lv D, Ren R, Xiaokaiti Y, Ma X, Zhao X, et al. Neuroprotective Effects of a Standardized Flavonoid Extract from Safflower against a Rotenone-Induced Rat Model of Parkinson’s Disease. Mol Basel Switz. 2016;21(9).

Ramadan KS, Khalil OA, Danial EN, Alnahdi HS, Ayaz NO. Hypoglycemic and hepatoprotective activity of Rosmarinus officinalis extract in diabetic rats. J Physiol Biochem. 2013;69(4):779–83.

Allen DA, Yaqoob MM, Harwood SM. Mechanisms of high glucose-induced apoptosis and its relationship to diabetic complications. J Nutr Biochem. 2005;16(12):705–713.

Circu ML, Aw TY. Reactive oxygen species, cellular redox systems, and apoptosis. Free Radic Biol Med. 2010;48(6):749–62.

Lelkes E, Unsworth BR, Lelkes PI. Reactive oxygen species, apoptosis and alte1red NGF-induced signaling in PC12 pheochromocytoma cells cultured in elevated glucose: AnIn Vitro cellular model for diabetic neuropathy. Neurotox Res. 2001;3(2):189–203.

Slater AF, Stefan C, Nobel I, Orrenius S. The role of intracellular oxidants in apoptosis. Biochim Biophys Acta BBA-Mol Basis Dis. 1995;1271(1):59–62.

Rajendran P, Nandakumar N, Rengarajan T, Palaniswami R, Gnanadhas EN, Lakshminarasaiah U, et al. Antioxidants and human diseases. Clin Chim Acta. 2014;436:332–47.

Rasoulian B, Kheirandish F. Herbal Medicines: from Traditional Medicine to Modern Experimental Approaches. Herb Med J. 2017;1(2).

Li S, Chen G, Zhang C, Wu M, Wu S, Liu Q. Research progress of natural antioxidants in foods for the treatment of diseases. Food Sci Hum Wellness. 2014;3(3):110–6.

Aruoma OI, Spencer JPE, Rossi R, Aeschbach R, Khan A, Mahmood N, et al. An evaluation of the antioxidant and antiviral action of extracts of rosemary and Provencal herbs. Food Chem Toxicol. 1996;34(5):449–56.

Ghasemzadeh MR, Amin B, Mehri S, Mirnajafi-Zadeh SJ, Hosseinzadeh H. Effect of alcoholic extract of aerial parts of Rosmarinus officinalis L. on pain, inflammation and apoptosis induced by chronic constriction injury (CCI) model of neuropathic pain in rats. J Ethnopharmacol. 2016;194:117–30.

Machado DG, Bettio LEB, Cunha MP, Capra JC, Dalmarco JB, Pizzolatti MG, et al. Antidepressant-like effect of the extract of Rosmarinus officinalis in mice: involvement of the monoaminergic system. Prog Neuropsychopharmacol Biol Psychiatry. 2009;33(4):642–50.

Section
Original Article