Thymoquinone Ameliorates Indomethacin-Induced Gastric Ulcers in Rats: A Dose Response Study

Abstract

Nigella sativa L. essential oil is a widely used medicinal plant throughout the world. It is very popular in various traditional systems of medicine like Unani and Tibb, Ayurveda and Siddha. Acute and chronic use of non-steroidal anti-inflammatory drugs (NSAIDs) is considered a significant cause of peptic ulcers. Different doses (20-60 mg/kg body weight) of thymoquinone (TQ), the main constituent of Nigella sativa L. essential oil, have been shown to protect gastric tissue from NSAID-induced peptic ulcers. Researchers attributed the protective effect of TQ to its antioxidant properties. This study aimed to determine effective antiulcer dose range of TQ. In this study, we conducted a series of experiments to determine the optimal dosage of TQ in indomethacin-induced gastric ulcers in rats. Additionally, we investigated the effect of TQ on superoxide dismutase (SOD) activity, glutathione (GSH) levels, and malondialdehyde (MDA) levels. Our results showed that, when administered at 40 and 20 mg/kg body weight, TQ was ineffective in preventing indomethacin-induced gastric ulcers. Moreover, TQ itself induced gastric ulcers at 40 mg/kg dose. As the doses of TQ decreased, the protective effect increased. 0.5 mg/kg TQ provided the best protection in terms of gastric ulcer area and antioxidant parameters, having statistically the same result with famotidine. Low dose TQ is an efficient protector of indomethacin-induced gastric damage, and it significantly enhances antioxidant parameters of gastric tissue. High dose TQ administration does not produce any desirable effects.

Keywords

• Thymoquinone
• Rats
• Indomethacin-induced gastric ulcer
• SOD
• GSH
• MDA

References

1. 1. Lam SK. Differences in peptic ulcer between East and West. Best Practice & Research Clinical Gastroenterology. 2000;14(1):41-52.
2. 2. Rosenstock SJ, Jørgensen T, Bonnevie O, Andersen LP. Does Helicobacter pylori infection explain all socio‐economic differences in peptic ulcer incidence? Genetic and psychosocial markers for incident peptic ulcer disease in a large cohort of Danish adults. Scandinavian Journal of Gastroenterology. 2004;39(9):823-9.
3. 3. Lanas A, Chan FKL. Peptic ulcer disease. Lancet. 2017;390(10094):613-24.
4. 4. Palle S, Kanakalatha A, Kavitha CN. Gastroprotective and Antiulcer Effects of Celastrus paniculatus Seed Oil Against Several Gastric Ulcer Models in Rats. J Diet Suppl. 2018;15(4):373-85.
5. 5. Drini M. Peptic ulcer disease and non-steroidal anti-inflammatory drugs. Aust Prescr. 2017;40(3):91-3.
6. 6. Takeuchi K. Pathogenesis of NSAID-induced gastric damage: importance of cyclooxygenase inhibition and gastric hypermotility. World J Gastroenterol. 2012;18(18):2147-60.
7. 7. Ishtiaq M, Hanif W, Khan MA, Ashraf M, Butt AM. An ethnomedicinal survey and documentation of important medicinal folklore food phytonims of flora of Samahni valley, (Azad Kashmir) Pakistan. Pak J Biol Sci. 2007;10(13):2241-56.
8. 8. Khémiri I, Bitri L. Effectiveness of Opuntia ficus indica L. inermis Seed Oil in the Protection and the Healing of Experimentally Induced Gastric Mucosa Ulcer. Oxid Med Cell Longev. 2019;2019:1568720-.

9. 9. Khan SA, Khan AM, Karim S, Kamal MA, Damanhouri GA, Mirza Z. Panacea seed "Nigella": A review focusing on regenerative effects for gastric ailments. Saudi J Biol Sci. 2016;23(4):542-53.
10. 10. Ghahramanloo KH, Kamalidehghan B, Akbari Javar H, Teguh Widodo R, Majidzadeh K, Noordin MI. Comparative analysis of essential oil composition of Iranian and Indian Nigella sativa L. extracted using supercritical fluid extraction and solvent extraction. Drug Des Devel Ther. 2017;11:2221-6.
11. 11. Abdelwahab SI, Sheikh BY, Taha MM, How CW, Abdullah R, Yagoub U, et al. Thymoquinone-loaded nanostructured lipid carriers: preparation, gastroprotection, in vitro toxicity, and pharmacokinetic properties after extravascular administration. Int J Nanomedicine. 2013;8:2163-72.
12. 12. Arslan SO, Gelir E, Armutcu F, Coskun O, Gurel A, Sayan H, et al. The protective effect of thymoquinone on ethanol-induced acute gastric damage in the rat. Nutr Res. 2005;25(7):673-80.
13. 13. El-Abhar HS, Abdallah DM, Saleh S. Gastroprotective activity of Nigella sativa oil and its constituent, thymoquinone, against gastric mucosal injury induced by ischaemia/reperfusion in rats. J Ethnopharmacol. 2003;84(2-3):251-8.
14. 14. Ahmad A, Husain A, Mujeeb M, Khan SA, Najmi AK, Siddique NA, et al. A review on therapeutic potential of Nigella sativa: A miracle herb. Asian Pac J Trop Biomed. 2013;3(5):337-52.
15. 15. 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(4):217-24.
16. 16. Asaduzzaman Khan M, Tania M, Fu S, Fu J. Thymoquinone, as an anticancer molecule: from basic research to clinical investigation. Oncotarget. 2017;8(31):51907-19.
17. 17. Zubair H, Khan HY, Sohail A, Azim S, Ullah MF, Ahmad A, et al. Redox cycling of endogenous copper by thymoquinone leads to ROS-mediated DNA breakage and consequent cell death: putative anticancer mechanism of antioxidants. Cell Death Dis. 2013;4(6):e660.
18. 18. Sun Y, Oberley LW, Li Y. A simple method for clinical assay of superoxide dismutase. Clin Chem. 1988;34(3):497-500.
19. 19. Sedlak J, Lindsay RH. Estimation of total, protein-bound, and nonprotein sulfhydryl groups in tissue with Ellman's reagent. Anal Biochem. 1968;25(1):192-205.
20. 20. Ohkawa H, Ohishi N, Yagi K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem. 1979;95(2):351-8.
21. 21. Polat B, Suleyman H, Alp HH. Adaptation of rat gastric tissue against indomethacin toxicity. Chem Biol Interact. 2010;186(1):82-9.
22. 22. Yoshida N, Takemura T, Granger DN, Anderson DC, Wolf RE, McIntire LV, et al. Molecular determinants of aspirin-induced neutrophil adherence to endothelial cells. Gastroenterology. 1993;105(3):715-24.
23. 23. Kanter M, Demir H, Karakaya C, Ozbek H. Gastroprotective activity of Nigella sativa L oil and its constituent, thymoquinone against acute alcohol-induced gastric mucosal injury in rats. World J Gastroenterol. 2005;11(42):6662-6.
24. 24. Salmani JM, Asghar S, Lv H, Zhou J. Aqueous solubility and degradation kinetics of the phytochemical anticancer thymoquinone; probing the effects of solvents, pH and light. Molecules. 2014;19(5):5925-39.
25. 25. Marsik P, Kokoska L, Landa P, Nepovim A, Soudek P, Vanek T. In vitro inhibitory effects of thymol and quinones of Nigella sativa seeds on cyclooxygenase-1- and -2-catalyzed prostaglandin E2 biosyntheses. Planta Med. 2005;71(8):739-42.
26. 26. Ragheb A, Attia A, Eldin WS, Elbarbry F, Gazarin S, Shoker A. The protective effect of thymoquinone, an anti-oxidant and anti-inflammatory agent, against renal injury: a review. Saudi J Kidney Dis Transpl. 2009;20(5):741-52.
27. 27. Kaseb AO, Chinnakannu K, Chen D, Sivanandam A, Tejwani S, Menon M, et al. Androgen receptor and E2F-1 targeted thymoquinone therapy for hormone-refractory prostate cancer. Cancer Res. 2007;67(16):7782-8.
28. 28. Badary OA. Thymoquinone attenuates ifosfamide-induced Fanconi syndrome in rats and enhances its antitumor activity in mice. J Ethnopharmacol. 1999;67(2):135-42.
29. 29. Abdel-Fattah AM, Matsumoto K, Watanabe H. Antinociceptive effects of Nigella sativa oil and its major component, thymoquinone, in mice. Eur J Pharmacol. 2000;400(1):89-97.
30. 30. Bayir Y, Albayrak A, Can I, Karagoz Y, Cakir A, Suleyman H, et al. Nigella sativa as a potential therapy for the treatment of lung injury caused by cecal ligation and puncture-induced sepsis model in rats. Cell Mol Biol (Noisy-le-grand). 2012;58 Suppl:OL1680-7.
31. 31. Salem ML. Immunomodulatory and therapeutic properties of the Nigella sativa L. seed. Int Immunopharmacol. 2005;5(13-14):1749-70.
32. 32. Magdy MA, Hanan el A, Nabila el M. Thymoquinone: Novel gastroprotective mechanisms. Eur J Pharmacol. 2012;697(1-3):126-31.
33. 33. Martin MJ, Jimenez MD, Motilva V. New issues about nitric oxide and its effects on the gastrointestinal tract. Curr Pharm Design. 2001;7(10):881-908.
34. 34. Naito Y, Yoshikawa T, Yoshida N, Kondo M. Role of oxygen radical and lipid peroxidation in indomethacin-induced gastric mucosal injury. Dig Dis Sci. 1998;43(9 Suppl):30S-4S.
35. 35. Takeuchi K, Takehara K, Ohuchi T. Diethyldithiocarbamate, a superoxide dismutase inhibitor, reduces indomethacin-induced gastric lesions in rats. Digestion. 1996;57(3):201-9.
36. 36. Sigthorsson G, Crane R, Simon T, Hoover M, Quan H, Bolognese J, et al. COX-2 inhibition with rofecoxib does not increase intestinal permeability in healthy subjects: a double blind crossover study comparing rofecoxib with placebo and indomethacin. Gut. 2000;47(4):527-32.
37. 37. Badary OA, Taha RA, Gamal el-Din AM, Abdel-Wahab MH. Thymoquinone is a potent superoxide anion scavenger. Drug Chem Toxicol. 2003;26(2):87-98.
38. 38. Laporte JR, Carne X, Vidal X, Moreno V, Juan J. Upper gastrointestinal bleeding in relation to previous use of analgesics and non-steroidal anti-inflammatory drugs.Catalan Countries Study on Upper Gastrointestinal Bleeding. Lancet. 1991;337:85-9.
39. 39. Mohamed A, Shoker A, Bendjelloul F, Mare A, Alzrigh M, Benghuzzi H, et al. Improvement of experimental allergic encephalomyelitis (EAE) by thymoquinone; an oxidative stress inhibitor. Biomed Sci Instrum. 2003;39:440-5.
40. 40. Akkus I. Serbest radikaller ve fizyopatolojik etkileri. Konya: Mimoza Yayınları 1995.
41. 41. Abdul Hamid Z, Budin SB, Wen Jie N, Hamid A, Husain K, Mohamed J. Nephroprotective effects of Zingiber zerumbet Smith ethyl acetate extract against paracetamol-induced nephrotoxicity and oxidative stress in rats. J Zhejiang Univ Sci B. 2012;13(3):176-85.
42. 42. Ghosh A, Sil PC. Anti-oxidative effect of a protein from Cajanus indicus L against acetaminophen-induced hepato-nephro toxicity. J Biochem Mol Biol. 2007;40(6):1039-49.
43. 43. Yousef MI, Omar SA, El-Guendi MI, Abdelmegid LA. Potential protective effects of quercetin and curcumin on paracetamol-induced histological changes, oxidative stress, impaired liver and kidney functions and haematotoxicity in rat. Food Chem Toxicol. 2010;48(11):3246-61.
44. 44. Havakhah S, Sadeghnia HR, Hajzadeh MA, Roshan NM, Shafiee S, Hosseinzadeh H, et al. Effect of Nigella sativa on ischemia-reperfusion induced rat kidney damage. Iran J Basic Med Sci. 2014;17(12):986-92.
45. 45. Mollazadeh H, Hosseinzadeh H. The protective effect of Nigella sativa against liver injury: a review. Iran J Basic Med Sci. 2014;17(12):958-66.