Combination of Nigella sativa with Glycyrrhiza glabra and Zingiber officinale augments their protective effects on doxorubicin-induced toxicity in h9c2 cells

Document Type : Original Article

Authors

1 Pharmacological Research Center of Medicinal Plants, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

2 Pharmacological Research Center of Medicinal Plants, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran Department of Pharmacology, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran

Abstract

Objective(s):The use of doxorubicin (DOX) is limited by its dose-dependent cardio toxicity in which reactive Oxygen Species (ROS) play an important role in the pathological process. The aim of this study was to evaluate the protective effect of three medicinal plants, Nigella sativa (N), Glycyrrhiza glabra (G) and Zingiber officinale (Z), and their combination (NGZ), against DOX-induced apoptosis and death in H9c2 cells.
Materials and Methods: The cells were incubated with different concentrations of each extract or NGZ for 4 hr which continued in the presence or absence of 5µM doxorubicin for 24 hr. Cell viability and the apoptotic rate were determined using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium (MTT) and propidium iodide (PI) staining assays, respectively. The level of ROS and lipid peroxidation were measured by fluorimetric methods.
Results: Treatment with doxorubicin increased ROS generation, enhanced malondialdehyde (MDA) formation, and induced apoptosis. Co-treatment of the cells with each herb extract increased viability of cells dose-dependently with a maximum protection effect of about 30%, and their potencies were N>G>Z. The combination of the threshold dose of each extract (NGZ) produced a similar effect, which was increased dose-dependently to a maximum protection of 70%. These effects were correlated with the effects of NGZ on ROS and MDA.
Conclusion: All of the extracts have some protective effects against DOX-induced toxicity in cardiomyocytes with similar efficacies, but with different potencies. However, NGZ produced much higher protective effect via reducing oxidative stress and inhibiting of apoptotic induction processes. Further investigations are needed to determine the effects of NGZ on DOX chemotherapy.  

Keywords

References

1. Wu S, Ko YS, Teng MS, Ko YL, Hsu LA, Hsueh C, et al. Adriamycin-induced  cardiomyocyte  and endothelial cell apoptosis: In vitro and in vivo studies. J Mol Cell Cardiol 2002; 34:1595–1607.
2. Bryant  J,  Picot J, Levitt G, Sullivan I, Baxter L, Clegg A. Cardioprotection against the toxic effects of anthracyclines given to children with cancer: a  systematic review.  Health Technol Assess 2007; 11:1-84.
3. Goreja WG. Black Seed: Nature’s Miracle Remedy. New York: Amazing Herbs Press; 2003.
4. Al-Rowais NA. Herbal medicine in the treatment of diabetes mellitus. Saudi Med J 2002; 23:1327-1331.
5. Burits M, Bucar F. Antioxidant activity of Nigella sativa essential oil. Phytother Res 2000; 14:323-328.
6. Houghton PJ, Zarka R, de las Heras B, Hoult JR. Fixed oil of Nigella sativa and derived thymoquinone inhibit eicosanoid generation in leukocytes and membrane lipid peroxidation. Planta Med 1995; 61: 33-36.
7. Khalife KH, Lupidi G. Non enzymatic reduction of thymoquinone in physiological conditions.  Free Radic Res 2007; 41:153-161.
8. Kanter M, Coskun O, Uysal H. The antioxidative and antihistaminic effect of Nigella sativa and its major constituent, thymoquinone on ethanol-induced gastric mucosal damage. Arch Toxicol 2006; 80:217-224.
9. Murugesan M, Ragunath M, Prabu T, Nadanasabapathi S, Sakthivel M, Manju V. Protective role of black cumin (Nigella sativa) on isoproterenol induced myocardial infarction in rats. Int J Pharmacol and Clin Sci 2012; 1:45-53.
10. Ahmed MA, Hassanein KMA. Cardio protective effects of Nigella sativa oil on lead induced cardio toxicity: Anti-inflammatory and antioxidant mechanism. J Physiol Pathophysiol 2013; 4:72-80.
11. Ebru U, Burak U, Yusuf S, Reyhan B, Arif K, Faruk TH, et al. Cardioprotective effects of Nigella sativa oil on cyclosporine A-induced cardio toxicity in rats. Basic Clin Pharmacol Toxicol 2008; 103:574–580.
12. al-Shabanah OA, Badary OA, Nagi MN, al-Gharably NM, al-Rikabi AC, al-Bekairi AM. Thymoquinone protects against doxorubicin-induced cardio toxicity without compromising its antitumor activity. J Experimen Clin Cancer Res 1998; 17:193–198.
13. Effenberger-Neidnicht K, Schobert R. Combinatorial effects of thymoquinone on the anti-cancer activity of doxorubicin. Cancer Chemother Pharmacol 2011; 67:867–874.
14. Nagi MN, Mansour MA. Protective effect of thymoquinone against doxorubicin-induced cardio toxicity in rats: A possible mechanism of protection. Pharmacol Res 2000; 41:283–289.
15. Afzal M, Al-Hadidi D, Menon M, Pesek J, Dhami MSI. Ginger: an ethnomedical, chemical and Pharmacological review. Drug Metabol Drug Interact 2001; 18:159-190.
16. Ezeonu C, Egbuna P, Ezeanyika US, Nkwonta CG, Idoko ND. Antihepatotoxicity studies of crude extract of Zingiber officinale on CCl4 induced toxicityand comparison of the extract’s fraction D hepatoprotective capacity. Res J Med Sci 2011; 5:102–107.
17. Forouzan S, Bahmani M, Parsaei P Mohsenzadegan A, Gholami-Ahangaran M, Sadeghi E, et al. Anti-parasitic activites of Zingiber officinale methanolic extract on Limnatis Nilotica. Global Veterinaria 2012; 9:144–148.
18. Mostafa OM, Eid RA, Adly MA. Antischistosomal activity of ginger (Zingiber officinale) against Schistosoma mansoni harbored in C57BL/6 mice. Parasitology Res 2011; 109:395–403.
19. Karuppiah P, Rajaram S. Antibacterial effect of Allium sativum cloves and Zingiber officinale rhizomes against multiple-drug resistant clinical pathogens. Asian Pacific J Tropical Biomed 2012; 2:597–601.
20. Al-Amin ZM, Thomson M, Al-Qattan KK, Peltonen-Shalaby R, Ali M. Anti-diabetic and hypolipidaemic properties of ginger (Zingiber officinale) in streptozotocin-induced diabetic rats. Br J Nutrition 2006; 96:660–666.
21. Baliga MS, Haniadka R, Pereira MM, Thilakchand KR, Rao S, Arora R. Radioprotective effects of Zingiber officinale (ginger): past, present and future. Food Funct 2012; 3:714–723.
22. Nicoll R, Henein MY. Ginger (Zingiber officinale): a hot remedy for cardiovascular disease? Int J Cardiol 2009; 131:408–409.
23. Darvishzadeh-Mahani F, Esmaeili-Mahani S, Komeili G, Sheibani V, Zare L. Ginger (Zingiber officinale) prevents the development of morphine analgesic tolerance and physical dependence in rats. J Ethnopharmacol 2012; 141:901–907.
24. Agrawal AK, Rao CV, Sairam K, Joshi VK, Goel RK. Effect of Piper longum Linn, Zingiber officinale Linn and Ferula species on gastric ulceration and secretion in rats. Indian J Exp Biol 2000; 38:994-998.
25. Ghasemzadeh A, Jaafar HZ, Rahmat A. Antioxidant activities, total phenolics and flavonoids content in two varieties of Malaysia young ginger (Zingiber officinale Roscoe). Molecules 2010; 15:4324–4333.
26. Asl MN, Hosseinzadeh H. Review of pharmacological effects of Glycyrrhiza sp. and its bioactive compounds. Phytother Res 2008; 22:709–724.
27. Nakagawa K, Kishida H, Arai N, Nishiyama T, Mae T. Licorice flavonoids suppress abdominal fat accumulation and increase in blood glucose level in obese diabetic KK-A(y) mice. Biol Pharm Bull 2004; 27:1775–1778.
28. Lim WY, Chia YY, Liong SY, Ton SH, Kadir KA, Husain SN. Lipoprotein lipase expression, serum lipid and tissue lipid deposition in orally administered glycyrrhizic acid reated rats. Lipids Health Dis 2009; 8: 31.
29. Wittschier N, Faller G, Hensel A. Aqueous extracts and polysaccharides from liquorice roots (Glycyrrhiza glabra L.) inhibit adhesion of helicobacter pylori to human gastric mucosa. J Ethnopharmacol 2009; 125:218–223.
30. Racková L, Jancinová V, Petríková M, Drábiková K, Noslál’ R, Stefek M, et al. Mechanism of anti-inflammatory action of liquorice extract and glycyrrhizin. Nat Prod Res 2007; 21:1234–1241.
31. Bafna PA, Balaraman R. Antioxidant activity of DHC-1, an herbal formulation, in experimentally-induced cardiac and renal damage. Phytother Res 2005; 19:216–221.
32. Visavadiya NP, Soni B, Dalwadi N. Evaluation of antioxidant and antiatherogenic properties of Glycyrrhiza glabra root using in vitro models. Int J Food Sci Nutr 2009; 60:135–149.
33. Ojha Sh, Golechha M, Kumari S, Bhatia J, Arya DS. Glycyrrhiza glabra protects from myocardial ischemia–reperfusion injury by improving hemodynamic, biochemical, histopathological and ventricular function. Exp Toxicol Pathol 2013; 65:219–227.
34. Choi HJ, Seon MR, Lim SS, Kim JS, Chun HS, Park JHY. Hexane/ethanol extract of Glycyrrhiza uralensis licorice suppresses doxorubicin-induced apoptosis in H9C2 rat cardiac myoblasts. Exp Biol Med (Maywood) 2008; 233:1554–1560.
35. Zhang L, Yang Y, Yu L, Wang Y, Liu L, Fan X. Cardioprotective effects of Glycyrrhiza uralensis extract against doxorubicin-induced toxicity. Int J Toxicol 2011; 30:181-189.
36. Parisella ML, Angelone T, Gattuso A, Cerra MC, Pellegrino D. Glycyrrhizin and glycyrrhetinic acid directly modulate rat cardiac performance. J Nutrition Biochem 2012; 23:69-75.
37. Sadeghnia HR, Yousefsani BS, Rashidfar M, Boroushaki MT, Assadpour E,   Ghorbani A. Protective effect of rutin on hexachlorobutadiene-induced nephrotoxicity. Ren Fail 2013; 35:1151-1155.
38. Mortazavian SM, Ghorbani A, Hesari TG. Effect of hydro-alcoholic extracts of viola tricolor and its fractions on proliferation of cervix carcinoma cells. Iran J Obstet Gynecol Infertil 2012; 15:9-16.
39. Tsutsui H, Kinugawa S, Matsushima S. Oxidative stress and heart failure. Am J Physiol Heart Circ Physiol 2011; 301:H2181-H2190.
40. Li H, Horke S, Forstermann U. Oxidative stress in vascular disease and its pharmacological prevention. Trends in Pharmacol Sci  2013; 34: 313-319.
41. Ahmed Z, Tang WH. Pharmacologic strategies to target oxidative stress in heart failure. Curr Heart Fail Rep 2012; 9:14-22.
42. Kang YJ, Chen Y, Epstein PN. Suppression of doxorubicin cardio toxicity by overexpression of catalase in the heart of transgenic mice. J Biol Chem 1996; 271:12610-12616.
43. Sheng R, Gu ZL, Xie ML, Zhou WX, Guo CY. Epigallocatechin gallate protects H9c2 cardiomyoblasts against hydrogen dioxides-induced apoptosis and telomere attrition. Eur J Pharmacol 2010; 641: 199-206.
44. Winstead MW, Lucas KK, Dennis EA. Group IV cytosolic phospholipase A2 mediates arachidonic acid release in H9c2 rat cardiomyocyte cells in response to hydrogen peroxide. Prostaglandins Other Lipid Mediat 2005; 78:55-66.
45. El-Daly ES. Protective effect of cysteine and vitamin E, Crocus sativus and Nigella sativa extracts on cisplatin-induced toxicity in rats. J Pharmacol (Belgium) 1998; 53:87–93.
46. El-Bakly WM, Louka ML, El-Halawany AM, Schaalan MF. 6-gingerol ameliorated doxorubicin-induced cardio toxicity: role of nuclear factor kappa B and protein glycation. Cancer Chemother Pharmacol 2012; 70:833-841.
47. Hibasami H, Iwase H, Yoshioka K, Takahashi H. Glycyrrhizin induces apoptosis in human stomach cancer KATO III and human promyelotic leukemia HL-60 cells. Int J Mol Med 2005; 16:233–236.
48. Hibasami H, Iwase H, Yoshioka K, Takahashi H. Glycyrrhetinic acid (a metabolic substance and aglycon of glycyrrhizin) induces apoptosis in human hepatoma, promyelotic leukemia and stomach cancer cells. Int J Mol Med 2006; 17:215–219.
49. Lee CS, Kim YJ, Lee MS, Hana ES, Lee SJ. 18β-
Glycyrrhetinic acid induces apoptotic cell death in SiHa cells and exhibits a synergistic effect against antibiotic anti-cancer drug toxicity. Life Sci 2008; 83:481–489.
50. Shukla Y, Singh M. Cancer preventive properties of ginger: a brief review. Food Chem Toxicol 2007; 45:683–690.
51. Jagetia GC, Reddy TK, Malagi KJ, Nayak BS, Naidu MBR, Ravikiran PB, et al. Antarth, a polyhedral preparation protects against the doxorubin-induced toxicity without compromising its antineoplastic activity. Phytother Res 2005; 19:772–778.
52. Liu L, Zhou HY, Ran K, Wang JB. Glycyrrhiznatis ameliorates rabbit myocardial ischemia-reperfusion injury through P38MAPK pathway. Nan Fang Yi Ke Da Xue Xue Bao 2010; 30:298–300.
Iranian Journal of Basic Medical Sciences
Volume 17, Issue 12 - Serial Number 12
December 2014
Pages 993-1000

Files

Share

How to cite

Statistics