Protective Effect of Diosgenin against H2O2-Induced Oxidative Stress on H9C2 Cells

Document Type : Research Articles

Authors

• Samaneh Jamshidi ¹
• Mehrdad Lahouti ¹
• Mohammad Taher Boroushaki ²
• Ali Ganjeali ¹
• Ahmad Ghorbani ²
• Mehdi Bihamta toosi ²

¹ Department of Biology, Faculty of Sciences, Ferdowsi University of Mashhad, Iran

² Department of Pharmacology, Faculty of Medicine, Mashhad University of Medical Science, Mashhad, Iran

https://doi.org/10.22067/jcmr.v0i0.36148

Abstract

Diosgenin is an important compound in pharmaceutical industry. It has various effects such as hypocholesterolemic action or antioxidant activity in HIV infected patients. Biological oxidation pathways are involved in causing or aggravating heart disease. This study investigated the potential protective effect of diosgenin on cell viability and antioxidant defenses of cultured H9C2 cells submitted to oxidative stress induced by H2O2. Viability of cells exposed to H2O2 was detected by MTT assay. The generation of ROS and hydrogen peroxide release after H2O2 were detected using the fluorescent probe H2DCF-DA. The lipid peroxidation product i.e. MDA formation was estimated by assessing the levels of thio-barbituric acid reactive substances (TBARS) using spectrophotometry. SOD activity was assayed with NWLSS (TM) Superoxide Dismutase (SOD) activity assay kit. Pretreatment of cells with 3-25 µM of diosgenin for 24 h before applying H2O2 completely prevented cell damage and significantly enhanced viability of H9C2 cells. Increased ROS induced by H2O2 was dose dependently prevented when cells were pretreated for 24 h with diosgenin. The level of the lipid peroxidation was significantly higher in H9C2 cells exposed to H2O2 as compared to the control and cells pretreated with diosgenin. SOD activity in cells treated with diosgenin significantly decreased compared with cells exposed to H2O2. These results show that treatment of H9C2 cells with diosgenin (3-25 µM) confers a significant protection against oxidative stress.

Keywords

• Diosgenin
• H9C2 cells
• oxidative stress
• MDA
• Cell viability

20.1001.1.20089147.2016.8.2.3.3

References

1. Accatino L., Pizarro M., Solis N. and Koenig C. S. (1998) Effects of diosgenin, a plant-derived steroid, on bile secretion and hepatocellular cholestasis induced by estrogens in the rat. Hepatology (Baltimore, Md.) 28:129-140.

2. Antunes F. and Cadenas E. (2001) Cellular titration of apoptosis with steady state concentrations of H(2)O(2): submicromolar levels of H(2)O(2) induce apoptosis through fenton chemistry independent of the cellular thiol state. Free Radical Biology and Medicine 30:1008-1018.

3. Aradhana, Rao A. R. and Kale R. K. (1992) Diosgenin--a growth stimulator of mammary gland of ovariectomized mouse. Indian Journal of Experimental Biology 30:367-370.

4. Attele A. S., Wu J. A. and Yuan C.-S. (1999) Ginseng pharmacology: Multiple constituents and multiple actions. Biochemical Pharmacology 58:1685-1693.

5. Beneytout J. L., Nappez C., Leboutet M. J. and Malinvaud G. (1995) A plant steroid, diosgenin, a new megakaryocytic differentiation inducer of HEL cells. Biochemical and Biophysical Research Communications 207:398-404.

6. Bernabucci U., Ronchi B., Lacetera N. and Nardone A. (2002) Markers of oxidative status in plasma and erythrocytes of transition dairy cows during hot season. Journal of Dairy Science 85:2173-2179.

7. Corbiere C., Liagre B., Bianchi A., Bordji K., Dauca M., Netter P. and Beneytout J. L. (2003) Different contribution of apoptosis to the antiproliferative effects of diosgenin and other plant steroids, hecogenin and tigogenin, on human 1547 osteosarcoma cells. International Journal of Oncology 22:899-905.

8. Davi G., Falco A. and Patrono C. (2005) Lipid peroxidation in diabetes mellitus. Antioxidants and Redox Signaling 7:256-268.

9. Kaneko M., Panagia V., Paolillo G., Majumder S., Ou C. and Dhalla N. S. (1990) Inhibition of cardiac phosphatidylethanolamine N-methylation by oxygen free radicals. Biochimica et Biophysica Acta 1021:33-38.

10. Kim D. S., Jeon B. K., Lee Y. E., Woo W. H. and Mun Y. J. (2012) Diosgenin induces apoptosis in HepG2 cells through generation of reactive oxygen species and mitochondrial pathway. Evidence-based complementary and alternative medicine: eCAM 2012:981675.

11. Liu M. J., Wang Z., Ju Y., Wong R. N. and Wu Q. Y. (2005) Diosgenin induces cell cycle arrest and apoptosis in human leukemia K562 cells with the disruption of Ca2+ homeostasis. Cancer Chemotherapy and Pharmacology 55:79-90.

12. Lores Arnaiz S., Travacio M., Llesuy S. and Rodriguez de Lores Arnaiz G. (1998) Regional vulnerability to oxidative stress in a model of experimental epilepsy. Neurochemical Research 23:1477-1483.

13. Moalic S., Liagre B., Corbiere C., Bianchi A., Dauca M., Bordji K. and Beneytout J. L. (2001) A plant steroid, diosgenin, induces apoptosis, cell cycle arrest and COX activity in osteosarcoma cells. FEBS Lett 506:225-230.

14. P M. K., ey, Mittra P. and Maheshwari P. K. (2013) Oxidative stress in epilepsy with comorbid psychiatric illness. National Journal of Physiology, Pharmacy and Pharmacology 3:92-96.

15. Peskin A. V. and Winterbourn C. C. (2000) A microtiter plate assay for superoxide dismutase using a water-soluble tetrazolium salt (WST-1). Clinica Chimica Acta; International Journal of Clinical Chemistry 293:157-166.

16. Pizarro J. G., Folch J., Vazquez De la Torre A., Verdaguer E., Junyent F., Jordan J., Pallas M. and Camins A. (2009) Oxidative stress-induced DNA damage and cell cycle regulation in B65 dopaminergic cell line. Free Radical Research 43:985-994.

17. Riley P. A. (1994) Free radicals in biology: oxidative stress and the effects of ionizing radiation. International Journal of Radiation Biology 65:27-33.

18. Roman I. D., Thewles A. and Coleman R. (1995) Fractionation of livers following diosgenin treatment to elevate biliary cholesterol. Biochimica et Biophysica Acta 1255:77-81.

19. Santanam N., Ramachandran S. and Parthasarathy S. (1998) Oxygen radicals, antioxidants, and lipid peroxidation. Seminars in Reproductive Endocrinology 16:275-280.

20. Srinivasan S., Koduru S., Kumar R., Venguswamy G., Kyprianou N. and Damodaran C. (2009) Diosgenin targets Akt-mediated prosurvival signaling in human breast cancer cells. International Journal of Cancer. Journal International du Cancer 125:961-967.

21. Stone J. R. and Yang S. (2006) Hydrogen peroxide: a signaling messenger. Antioxidants & Redox Signaling 8:243-270.

22. Thannickal V. J. and Fanburg B. L. (2000) Reactive oxygen species in cell signaling. American journal of physiology. Lung Cellular and Molecular Physiology 279:L1005-1028.

23. Toyokuni S. and Akatsuka S. (2007) Pathological investigation of oxidative stress in the post-genomic era. Pathology International 57:461-473.

24. Trevisan M., Browne R., Ram M., Muti P., Freudenheim J., Carosella A. M. and Armstrong D. (2001) Correlates of markers of oxidative status in the general population. American Journal of Epidemiology 154:348-356.

25. Turchan J., Pocernich C. B., Gairola C., Chauhan A., Schifitto G., Butterfield D. A., Buch S., Narayan O., Sinai A., Geiger J., Berger J. R., Elford H. and Nath A. (2003) Oxidative stress in HIV demented patients and protection ex vivo with novel antioxidants. Neurology 60:307-314.

26. Uchida K., Shiraishi M., Naito Y., Torii Y., Nakamura Y. and Osawa T. (1999) Activation of stress signaling pathways by the end product of lipid peroxidation. 4-hydroxy-2-nonenal is a potential inducer of intracellular peroxide production. The Journal of Biological Chemistry 274:2234-2242.

27. Ueda Y., Yokoyama H., Ohya-Nishiguchi H. and Kamada H. (1998) ESR spectroscopy for analysis of hippocampal elimination of a nitroxide radical during kainic acid-induced seizure in rats. Magnetic Resonance in Medicine: 40:491-493.

28. Wang S. L., Cai B., Cui C. B., Liu H. W., Wu C. F. and Yao X. S. (2004) Diosgenin-3-O-alpha-L-rhamnopyranosyl-(1 --> 4)-beta-D-glucopyranoside obtained as a new anticancer agent from Dioscorea futschauensis induces apoptosis on human colon carcinoma HCT-15 cells via mitochondria-controlled apoptotic pathway. Journal of Asian Natural Products Research 6:115-125.

29. Wei M. C., Zong W. X., Cheng E. H., Lindsten T., Panoutsakopoulou V., Ross A. J., Roth K. A., MacGregor G. R., Thompson C. B. and Korsmeyer S. J. (2001) Proapoptotic BAX and BAK: a requisite gateway to mitochondrial dysfunction and death. Science (New York, N.Y.) 292:727-730.

30. Yagi K. (1987) Lipid peroxides and human diseases. Chemistry and Physics of Lipids 45:337-351.

31. Zaleska M. M. and Wilson D. F. (1989) Lipid Hydroperoxides Inhibit Reacylation of Phospholipids in Neuronal Membranes. Journal of Neurochemistry 52:255-260.