Effects of Nigella sativa oil and ascorbic acid against oxytetracycline-induced hepato-renal toxicity in rabbits

Document Type: Original Article


1 Pharmacology Department, Faculty of Veterinary Medicine, Suez Canal University, Ismailia, Egypt

2 Clinical Pathology Department, Faculty of Veterinary Medicine, Kafrelsheikh University, Kafr El-Sheikh, Egypt


Objective(s):  Oxytetracycline (OTC) is a broad spectrum antibiotic widely used for treatment of a wide range of infections. However, its improper human and animal use leads to toxic effects, including hepatonephrotoxicity. Our objective was to evaluate protective effects of Nigella sativa oil (NSO) and/or ascorbic acid (AA), against OTC-induced hepatonephrotoxicity in rabbits.
Materials and Methods: Forty male white New Zealand rabbits were divided into 5 groups of eight each. The 1st group (control) was given saline. The 2nd group was given OTC (200 mg/kg, orally). The 3rd and 4th groups were orally administered NSO and AA (2 ml/kg and 200 mg/kg respectively) 1 hr before OTC administration at the same dose regimen used for the 2nd group. Both NSO and AA were given in combination for the 5th group along with OTC administration. Serum biochemical parameters related to liver and kidney injury were evaluated, and lipid peroxidation as well as antioxidant markers in hepatic and renal tissues were examined.
Results: OTC-treated animals revealed significant alterations in serum biochemical hepato-renal injury markers, and showed a markedly increase in hepato-renal lipid peroxidation and inhibition in tissue antioxidant biomarkers. NSO and AA protect against OTC-induced serum and tissue biochemical alterations when each of them is used alone or in combination along with OTC treatment. Furthermore, both NSO and AA produced synergetic hepatoprotective and antioxidant properties.
Conclusion: The present study revealed the preventive role of NSO and/or AA against the toxic effects of OTC through their free radical-scavenging and potent antioxidant activities.


1.   Nelson ML, Levy SB. The history of the tetracyclines. Ann N Y Acad Sci 2011; 1241:17-32.

2.   Polec RB, Yeh SD, Shils ME. Protective effect of ascorbic acid, isoascorbic acid and mannitol against tetracycline-induced nephrotoxicity. J Pharmacol Exp Ther 1971; 178:152-158.

3.   Naseer F, Alam M. The protective effect of ascorbic acid on oxytetracycline induced nephrotoxicity and hepatotoxicity. J Pak Med Assoc 1987; 37:73-75.

4.   Saraswat B, Visen PK, Patnaik GK, Dhawan BN. Protective effect of picroliv, active constituent of Picrorhiza kurrooa, against oxytetracycline induced hepatic damage. Indian J Exp Biol 1997; 35:1302-1305.

5.   Ibrahim AE, Abdel-Daim MM. Modulating effects of Spirulina platensis against tilmicosin-induced cardiotoxicity in mice. Cell Journal  Cell J. 2015; 17(1): 137-144.

6.   Madkour FF, Abdel-Daim MM. Hepatoprotective and Antioxidant Activity of Dunaliella salina in Paracetamol-induced Acute Toxicity in Rats. Indian J Pharm Sci 2013; 75:642-648.

7.   Abdel-Daim MM, Abd Eldaim MA, Mahmoud MM. Trigonella foenum-graecum protection against deltamethrin-induced toxic effects on haematological, biochemical, and oxidative stress parameters in rats. Can J Physiol Pharmacol 2014; 92:679-685.

8.   Abdel-Daim MM, Abuzead SM, Halawa SM. Protective role of Spirulina platensis against acute deltamethrin-induced toxicity in rats. PLoS One 2013; 8:e72991.

9.   Abdel-Daim MM. Pharmacodynamic interaction of Spirulina platensis with erythromycin in Egyptian Baladi bucks (Capra hircus). Small Ruminant Res 2014; 120:234-241.

10. Lutterodt H, Luther M, Slavin M, Yin JJ, Parry J, Gao JM, et al. Fatty acid profile, thymoquinone content, oxidative stability, and antioxidant properties of cold-pressed black cumin seed oils. Lwt-Food Sci Technol 2010; 43:1409-1413.

11. Kanter M, Coskun O, Budancamanak M. Hepatoprotective effects of Nigella sativa L and Urtica dioica L on lipid peroxidation, antioxidant enzyme systems and liver enzymes in carbon tetrachloride-treated rats. World J Gastroentero 2005; 11:6684-6688.

12. Abdel-Wahhab MA, Aly SE. Antioxidant property of Nigella sativa (black cumin) and Syzygium aromaticum (clove) in rats during aflatoxicosis. J Appl Toxicol 2005; 25:218-223.

13. Mansour MA, Ginawi OT, El-Hadiyah T, El-Khatib AS, Al-Shabanah OA, Al-Sawaf HA. Effects of volatile oil constituents of Nigella sativa on carbon tetrachloride-induced hepatotoxicity in mice: Evidence for antioxidant effects of thymoquinone. Res Commun Mol Pathol Pharmacol. 2001;110:239–51.

14. Barakat MK, Oda NR, Bayoumy FA, Bayoumy FA. Effect of Nigella sativa on Carbon Tetrachloride and Paracetamol Induced hepatotoxicity: Role of Antioxidant Enzymes and Cytokines. Faseb J 2010; 24.

15. El-Demerdash FM, Yousef MI, Zoheir MA. Stannous chloride induces alterations in enzyme activities, lipid peroxidation and histopathology in male rabbit: antioxidant role of vitamin C. Food Chem Toxicol 2005; 43:1743-1752.

16. Kojo S. Vitamin C: basic metabolism and its function as an index of oxidative stress. Curr Med Chem 2004; 11:1041-1064.

17. Carr AC, Frei B. Toward a new recommended dietary allowance for vitamin C based on antioxidant and health effects in humans. Am J Clin Nutr 1999; 69:1086-1107.

18. Grosicki A. Influence of vitamin C on cadmium absorption and distribution in rats. J Trace Elem Med Biol 2004; 18:183-187.

19. Sen Gupta R, Sen Gupta E, Dhakal BK, Thakur AR, Ahnn J. Vitamin C and vitamin E protect the rat testes from cadmium-induced reactive oxygen species. Mol Cells 2004; 17:132-139.

20. Shalan MG, Mostafa MS, Hassouna MM, El-Nabi SE, El-Refaie A. Amelioration of lead toxicity on rat liver with Vitamin C and silymarin supplements. Toxicology 2005; 206:1-15.

21. Jayanthi R, Subash P. Antioxidant effect of caffeic Acid on oxytetracycline induced lipid peroxidation in albino rats. Indian J Clin Biochem 2010; 25:371-375.

22. Saleem U, Ahmad B, Rehman K, Mahmood S, Alam M, Erum A. Nephro-protective effect of vitamin C and Nigella sativa oil on gentamicin associated nephrotoxicity in rabbits. Pak J Pharm Sci 2012; 25:727-730.

23. Reitman S, Frankel S. A colorimetric method for the determination of serum glutamic oxalacetic and glutamic pyruvic transaminases. Am J Clin Pathol 1957; 28:56-63.

24. Tietz NW, Burtis CA, Duncan P, Ervin K, Petitclerc CJ, Rinker AD, et al. A reference method for measurement of alkaline phosphatase activity in human serum. Clin Chem 1983; 29:751-761.

25. Richmond W. Preparation and properties of a cholesterol oxidase from Nocardia sp. and its application to the enzymatic assay of total cholesterol in serum. Clin Chem 1973; 19:1350-1356.

26. Allain CC, Poon LS, Chan CS, Richmond W, Fu PC. Enzymatic determination of total serum cholesterol. Clin Chem 1974; 20:470-475.

27. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951; 193:265-275.

28. Schattmann K. A spectrophotometric quantitative caffein-free method for determination of the serum bilirubin index with the Lange universal colorimeter. Arztl Wochensch 1952; 7:1154-1156.

29. Buhl SN, Jackson KY. Optimal conditions and comparison of lactate dehydrogenase catalysis of the lactate-to-pyruvate and pyruvate-to-lactate reactions in human serum at 25, 30, and 37 degrees C. Clin Chem 1978; 24:828-831.

30. Coulombe JJ, Favreau L. A new simple semimicro method for colorimetric determination of urea. Clin Chem 1963; 9:102-108.

31. Whitehead TP, Bevan EA, Miano L, Leonardi A. Defects in diagnostic kits for determination of urate in serum. Clin Chem 1991; 37:879-881.

32. Larsen K. Creatinine assay in the presence of protein with LKB 8600 Reaction Rate Analyser. Clin Chim Acta 1972; 38:475-476.

33. Mihara M, Uchiyama M. Determination of malonaldehyde precursor in tissues by thiobarbituric acid test. Anal Biochem 1978; 86:271-278.

34. Aebi H. Catalase in vitro. Methods Enzymol 1984; 105:121-126.

35. Nishikimi M, Appaji N, Yagi K. The occurrence of superoxide anion in the reaction of reduced phenazine methosulfate and molecular oxygen. Biochem Biophys Res Commun 1972; 46:849-854.

36. Beutler E, Duron O, Kelly BM. Improved method for the determination of blood glutathione. J Lab Clin Med 1963; 61:882-888.

37. Koracevic D, Koracevic G, Djordjevic V, Andrejevic S, Cosic V. Method for the measurement of antioxidant activity in human fluids. J Clin Pathol 2001; 54:356-361.

38. Halling-Sorensen B, Nors Nielsen S, Lanzky PF, Ingerslev F, Holten Lutzhoft HC, Jorgensen SE. Occurrence, fate and effects of pharmaceutical substances in the environment--a review. Chemosphere 1998; 36:357-393.

39. Southwood LL. Principles of antimicrobial therapy: what should we be using? Vet Clin North Am Equine Pract 2006; 22:279-296.