Abstract

Thymoquinone (THQ) reported to have beneficial effects on bone disorders. Thus the present study was designed to investigate its protective effects on experimentally-induced osteoporosisin ovariectomized (OVX) rats. The OVX rats were treated with different doses (2.5, 5 and 10 mg/kg/day) of THQ for ten weeks. Bone turnover biomarkers and inflammatory cytokines were determined using ELISA techniques. Changes in bones cortical and trabecular morphometric parameters were determined using micro-CT scan. Body weights and abdominal fats were significantly increased in OVX rats while the uterus and femoral bone weights were markedly inhibited in OVX rats. Estrogen deficiency elevated the level of bone turnover biomarkers as well as inflammatory cytokines in serum. Moreover, cortical and trabecular morphometric parameters of the femoral bones were severely altered by ovariectomy operation.THQ treatments inhibited the increased bone turnover biomarkers and inflammatory cytokines in dose dependent manner. The altered morphometric parameters of the femoral bones were also markedly attenuated by THQ. Present data revealed that THQ has osteoprotective properties may be due to its ability to inhibit osteoclastogenesis inducing factors and inflammation, which stabilize the osteoclasts and decreased their bone matrix's degradation or resorption. We believe that this effect may be through its both the anti-inflammatory and antioxidant properties.

Keywords:

Inflammatory mediators, micro CT, ovariectomy, osteoclastogensis, postmenopausal osteoporosis, thymoquinone,

References

1. Agarwal, R., M.D. Kharya and R. Shrivastava, 1979. Antimicrobial and anthelmintic activities of the essential oil of Nigella sativa Linn. Indian J. Exp. Biol., 17: 1264-1265.
   PMid:549848    
2. Ahmad, S. and Z.H. Beg, 2013. Hypolipidemic and antioxidant activities of thymoquinone and limonene in atherogenic suspension fed rats. Food Chem., 138: 1116-1124.
   CrossRef    PMid:23411222    
3. Al-Majed, A.A., F.A. Al-Omar and M.N. Nagi, 2006. Neuroprotective effects of thymoquinone against transient forebrain ischemia in the rat hippocampus. Eur. J. Pharmacol., 543: 40-47.
   CrossRef    PMid:16828080    
4. Badary, O.A., A.B. Abdel-Naim, M.H. Abdel-Wahab and F.M. Hamada, 2000. The influence of thymoquinone on doxorubicin-induced hyperlipidemic nephropathy in rats. Toxicology, 143: 219-226.
   CrossRef    
5. Badary, O.A., M.F. Abd-Ellah, M.A. El-Mahdy, S.A. Salama and F.M. Hamada, 2007. Anticlastogenic activity of thymoquinone against benzo(a)pyrene in mice. Food Chem. Toxicol., 45: 88-92.
   CrossRef    PMid:17011106    
6. Badary, O.A., R.A. Taha, A.M. Gamal el-Din and M.H. Abdel-Wahab, 2003. Thymoquinone is a potent superoxide anion scavenger. Drug Chem. Toxicol., 26: 87-98.
   CrossRef    PMid:12816394    
7. Bai, X.C., D. Lu, J. Bai, H. Zheng, Z.Y. Ke, X.M. Li and S.Q. Luo, 2004. Oxidative stress inhibits osteoblastic differentiation of bone cells by ERK and NF-kappaB. Biochem. Biophys. Res. Commun., 314: 197-207.
   CrossRef    PMid:14715266    
8. Ceballos-Picot, I., J.M. Trivier, A. Nicole, P.M. Sinet and M. Thevenin, 1992. Age-correlated modifications of copper-zinc superoxide dismutase and glutathione-related enzyme activities in human erythrocytes. Clin. Chem., 38: 66-70.
   PMid:1733608    
9. El-Tahir, K.E., M.M. Ashour and M.M. Al-Harbi, 1993. The cardiovascular actions of the volatile oil of the black seed (Nigella sativa) in rats: Elucidation of the mechanism of action. Gen. Pharmacol., 24: 1123-1131.
   CrossRef    
10. El-Abhar, H.S., D.M. Abdallah and S. Saleh, 2003. Gastroprotective activity of Nigella sativa oil and its constituent, thymoquinone, against gastric mucosal injury induced by ischaemia/reperfusion in rats. J. Ethnopharmacol., 84: 251-258.
    CrossRef    
11. El-Mahmoudy, A., H. Matsuyama, M.A. Borgan, Y. Shimizu, M.G. El-Sayed, N. Minamoto and T. Takewaki, 2002. Thymoquinone suppresses expression of inducible nitric oxide synthase in rat macrophages. Int. Immunopharmacol., 2: 1603-1611.
    CrossRef    
12. Fararh, K.M., Y. Shimizu, T. Shiina, H. Nikami, M.M. Ghanem and T. Takewaki, 2005. Thymoquinone reduces hepatic glucose production in diabetic hamsters. Res. Vet. Sci., 79: 219-223.
    CrossRef    PMid:16054891    
13. Galli, C., G. Passeri and G.M. Macaluso, 2011. FoxOs, Wnts and oxidative stress-induced bone loss: New players in the periodontitis arena? J. Periodontal. Res., 46: 397-406.
    CrossRef    PMid:21332475    
14. Grases, F., P. Sanchis, R.M. Prieto, J. Perello and A.A. Lopez-Gonzalez, 2010. Effect of tetracalcium dimagnesium phytate on bone characteristics in ovariectomized rats. J. Med. Food, 13: 1301-1306.
    CrossRef    PMid:21091244    
15. Ha, H., J.H. Lee, H.N. Kim and Z.H. Lee, 2011. alpha-Tocotrienol inhibits osteoclastic bone resorption by suppressing RANKL expression and signaling and bone resorbing activity. Biochem. Biophys. Res. Commun., 406: 546-551.
    CrossRef    PMid:21352805    
16. Hajhashemi, V., A. Ghannadi and H. Jafarabadi, 2004. Black cumin seed essential oil, as a potent analgesic and antiinflammatory drug. Phytother Res., 18: 195-199.
    CrossRef    PMid:15103664    
17. Henry, M.J., J.A. Pasco, G.C. Nicholson, E. Seeman and M.A. Kotowicz, 2000. Prevalence of osteoporosis in Australian women: Geelong osteoporosis study. J. Clin. Densitom, 3: 261-268.
    CrossRef    
18. Hertrampf, T., B. Schleipen, C. Offermanns, M. Velders, U. Laudenbach and P. Diel, 2009. Comparison of the bone protective effects of an isoflavone-rich diet with dietary and subcutaneous administrations of genistein in ovariectomized rats. Toxicol. Lett., 184: 198-203.
    CrossRef    PMid:19063953    
19. Houghton, P.J., R. Zarka, B. de las Heras and J.R. Hoult, 1995. Fixed oil of Nigella sativa and derived thymoquinone inhibit eicosanoid generation in leukocytes and membrane lipid peroxidation. Planta Med., 61: 33-36.
    CrossRef    PMid:7700988    
20. Kaczmarczyk-Sedlak, I., J. Folwarczna and H.I. Trzeciak, 2009. Thalidomide affects the skeletal system of ovariectomized rats. Pharmacol. Rep., 61: 529-538.
    CrossRef    
21. Kara, M.I., K. Erciyas, A.B. Altan, M. Ozkut, S. Ay and S. Inan, 2012. Thymoquinone accelerates new bone formation in the rapid maxillary expansion procedure. Arch Oral Biol., 57: 357-363.
    CrossRef    PMid:22036504    
22. Khosla, S., J.J. Westendorf and M.J. Oursler, 2008. Building bone to reverse osteoporosis and repair fractures. J. Clin. Invest., 118: 421-428.
    CrossRef    PMid:18246192 PMCid:PMC2214701    
23. Kirui, P.K., J. Cameron, H.A. Benghuzzi, M. Tucci, R. Patel, F. Adah and G. Russell, 2004. Effects of sustained delivery of thymoqiunone on bone healing of male rats. Biomed. Sci. Instrum., 40: 111-116.
    PMid:15133944    
24. Kruk, I., T. Michalska, K. Lichszteld, A. Kladna and H.Y. Aboul-Enein, 2000. The effect of thymol and its derivatives on reactions generating reactive oxygen species. Chemosphere, 41: 1059-1064.
    CrossRef    
25. Lean, J., B. Kirstein, Z. Urry, T. Chambers and K. Fuller, 2004. Thioredoxin-1 mediates osteoclast stimulation by reactive oxygen species. Biochem. Biophys. Res. Commun., 321: 845-850.
    CrossRef    PMid:15358104    
26. Lean, J.M., J.T. Davies, K. Fuller, C.J. Jagger, B. Kirstein, G.A. Partington, Z.L. Urry and T.J. Chambers, 2003. A crucial role for thiol antioxidants in estrogen-deficiency bone loss. J. Clin. Invest., 112: 915-923.
    CrossRef    PMid:12975476 PMCid:PMC193670    
27. Maggio, D., M. Barabani, M. Pierandrei, M.C. Polidori, M. Catani, P. Mecocci, U. Senin, R. Pacifici and A. Cherubini, 2003. Marked decrease in plasma antioxidants in aged osteoporotic women: Results of a cross-sectional study. J. Clin. Endocrinol. Metab., 88: 1523-1527.
    CrossRef    PMid:12679433    
28. Melhus, H., K. Michaelsson, A. Kindmark, R. Bergström, L. Holmberg, H. Mallmin, A. Wolk and S. Ljunghall, 1998. Excessive dietary intake of vitamin A is associated with reduced bone mineral density and increased risk for hip fracture. Ann Intern. Med., 129: 770-778.
    CrossRef    PMid:9841582    
29. Mertz, W., 1990. The role of trace elements in the aging process. Prog. Clin. Biol. Res., 326: 229-240.
    PMid:2405420    
30. Mohamed, A., D.M. Afridi, O. Garani and M. Tucci, 2005. Thymoquinone inhibits the activation of NF-kappaB in the brain and spinal cord of experimental autoimmune encephalomyelitis. Biomed. Sci. Instrum., 41: 388-393.
    PMid:15850137    
31. Mosekilde, L., 1990. Consequences of the remodelling process for vertebral trabecular bone structure: A scanning electron microscopy study (uncoupling of unloaded structures). Bone Miner., 10: 13-35.
    CrossRef    
32. Nagi, M.N. and M.A. Mansour, 2000. Protective effect of thymoquinone against doxorubicin-induced cardiotoxicity in rats: A possible mechanism of protection. Pharmacol. Res., 41: 283-289.
    CrossRef    PMid:10675279    
33. Nagi, M.N., K. Alam, O.A. Badary, O.A. Al-Shabanah, H.A. Al-Sawaf and A.M. Al-Bekairi, 1999. Thymoquinone protects against carbon tetrachloride hepatotoxicity in mice via an antioxidant mechanism. Biochem. Mol. Biol. Int., 47: 153-159.
    PMid:10092955    
34. Nian, H., M.H. Ma, S.S. Nian and L.L. Xu, 2009. Antiosteoporotic activity of icariin in ovariectomized rats. Phytomedicine, 16: 320-326.
    CrossRef    PMid:19269147    
35. Pasco, J.A., G.C. Nicholson, F. Ng, M.J. Henry, L.J. Williams, M.A. Kotowicz, J.M. Hodge, S. Dodd, F. Kapczinski, C.S. Gama and M. Berk, 2008. Oxidative stress may be a common mechanism linking major depression and osteoporosis. Acta Neuropsychiatrica, 20: 112-116.
    CrossRef    PMid:26951034    
36. Pasco, J.A., M.J. Henry, L.K. Wilkinson, G.C. Nicholson, H.G. Schneider and M.A. Kotowicz, 2006. Antioxidant vitamin supplements and markers of bone turnover in a community sample of nonsmoking women. J. Womens Health (Larchmt), 15: 295-300.
    CrossRef    PMid:16620188    
37. Potu, B.K., M.S. Rao, G.K. Nampurath, M.R. Chamallamudi, K. Prasad, S.R. Nayak, P.K. Dharmavarapu, V. Kedage and K.M. Bhat, 2009. Evidence-based assessment of antiosteoporotic activity of petroleum-ether extract of Cissus quadrangularis Linn. on ovariectomy-induced osteoporosis. Ups J. Med. Sci., 114: 140-148.
    CrossRef    PMid:19736603 PMCid:PMC2852762    
38. Rachner, T.D., S. Khosla and L.C. Hofbauer, 2011. Osteoporosis: Now and the future. Lancet, 377: 1276-1287.
    CrossRef    
39. Ramprasath, V.R., P. Shanthi and P. Sachdanandam, 2006. Curative effect of Semecarpus anacardium Linn. nut milk extract against adjuvant arthritis with special reference to bone metabolism. Chem. Biol. Interact., 160: 183-92.
    CrossRef    PMid:16513099    
40. Riggs, B.L., S. Khosla and L.J. Melton, 2002. Sex steroids and the construction and conservation of the adult skeleton. Endocr. Rev., 23: 279-302.
    CrossRef    PMid:12050121    
41. Rooney, S. and M.F. Ryan, 2005. Effects of alpha-hederin and thymoquinone, constituents of Nigella sativa, on human cancer cell lines. Anticancer Res., 25: 2199-204.
    PMid:16158964    
42. Sanders, K.M., M.A. Kotowicz and G.C. Nicholson, 2007. Potential role of the antioxidant N-acetylcysteine in slowing bone resorption in early post-menopausal women: A pilot study. Transl. Res., 150: 215.
    CrossRef    PMid:17900508    
43. Vaillancourt, F., P. Silva, Q. Shi, H. Fahmi, J.C. Fernandes and M. Benderdour, 2011. Elucidation of molecular mechanisms underlying the protective effects of thymoquinone against rheumatoid arthritis. J. Cell Biochem., 112: 107-117.
    CrossRef    PMid:20872780    
44. Worthen, D.R., O.A. Ghosheh and P.A. Crooks, 1998. The in vitro anti-tumor activity of some crude and purified components of blackseed, Nigella sativa L. Anticancer Res., 18: 1527-1532.
    PMid:9673365    

Competing interests

The authors have no competing interests.

Open Access Policy

This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.

Copyright

The authors have no competing interests.