Nigella sativa (Black seed) oil: Anti-inflammatory and antioxidant effects

1. Nigella sativa (Black seed) oil: Anti-inflammatory and antioxidant effects in experimental models of allergic asthma

2. Presented bySamira SalehProfessor of Pharmacology and ToxicologyFaculty of Pharmacy, Cairo UniversityCairo, Egypt

3. This work has been done in collaboration with E.M.S. Eldenshary L. Mahran N. Salah

4. Nigella sativa • Nigella sativa is an erect annual herb of the Mediterranean region and Asia. • The seeds are known as al habbah al sawdaa, Habbet el baraka, black seed, black cumin, Shung & others. • It is widely used in Egypt as a condiment and as a natural remedy for many ailments. • The expressed oil from the seeds is available in the market in the form of gelatin capsules used in folk medicine for treatment of many diseases, among which asthma & other inflammatory disorders.

5. Wheezing disappeared gradually in children with asthmatic bronchitis (Badr El-Din,1960) • Vital capacity was improved in asthmatics (Mahfouz et al ,1965 , 1969) • Pulmonary function (Abu Rayan et al, 1988 ) • Allergic airway inflammation was ameliorated by inhibiting T-cell proliferation in rats (Shahzadet al, 2009) • Lung remodeling, induced by repetitive allergen challenge in rats, was attenuated (Raza Asim et al, 2010)

6. قال رسول الله صلى الله عليه و سلم " عليكم بهذه الحبة السوداء ، فإن فيها شفاء من كلّ داء إلا السَّام ” رواه البخاري (5688) ومسلم (2215)

7. Background Inflammatory mediators play a central role in bronchial asthma. There is an interaction of many cell types & multiple mediators. In an allergic or immediate hypersensitivity response, mast cells are the primary effector cells involved. Airway inflammation is associated with an imbalance between formation of ROS at the inflammatory sites and endogenous defense.

8. Aim of the study The aim of this study is to evaluate the basis for the beneficial effect of Nigella oil in bronchial asthma by the use of hypersensitive guinea pigs and rats. • Immunological release of mediators from perfused lung preparations • Non-immunological release of histamine from mast cell • Antioxidant effect by measuring MDA, SOD & GSH

9. Methods • The isolated perfused lung preparation Histamine ProstanglandinsLeukotrienes • Rat peritoneal mast cells Histamine release by 48/80 • Antioxidant effects MDA, SOD, GSH

10. The perfused guinea pig lung Normal guinea pigs Sensitized guinea pigs Given saline 200 mg OV at day 0 In both groups, guinea pigs were allocated into 3 subgroups. Two were treated with Nigella oil, either 0.5 ml/kg or 2.5 ml/kg orally & the third was an untreated control. Treatment started on day 0, continued for 3 weeks Blood MDA SOD GSH Isolated perfused lungs Hm LTS PG2

11. The perfused guinea pig lung • The lung was cannulated through the pulmonary artery • It was perfused at a rate of 8 ml/min • Challenged with 10 mg OV in 1ml • The perfusate was collected 5 minutes before challenge and 5 minutes after challenge • Divided into 3 aliquots for determination of histamine LTC4/D4/E4/F4 PG2

12. 2. Rat peritoneal mast cell Rats were sensitized as follows: • First day: a mixture of OV & Bordetella pertussis vaccine s.c. • Second day: OV and Freund’s incomplete adjuvant • Third day: OV only BALF was collected 3 weeks after the first injection Compound 48/80 was used as a challenge to induce histamine release from mast cells

13. Results Effect of Nigella oil on mediators released from the normal lung Effect of Nigella oil on mediators released from the sensitized lung

14. Effect of Nigella oil on mediators released from isolated perfused lungsof normal guinea pigs Histamine Leukotrienes PGE2

15. +67% Histamine - 41% Normal

16. Leukotrienes -17% Normal

17. PGE2 +21% Normal

18. 2. Effect of Nigella oil on mediators released from isolated perfused lungsof sensitized guinea pigs Histamine Leukotrienes PGE2

19. Histamine +38% 35% - 46% Sensitized

20. Leukotrienes 52% +45% - 38% Sensitized

21. PGE2 + 21% + 37% Sensitized

22. II. Effect of Nigella oil on histamine release from rat peritoneal mast cells

23. -20% -39%

24. 62% -11% -28%

25. III. Antioxidant effects

26. Endogenous defense and free radicals There is an imbalance between formation of ROS at the inflammatory sites of the airways and endogenous defense

27. Comparison betweenblood MDA, SOD, GSHin normal & sensitized animals

28. +221%

29. +64%

30. +79%

31. Effect of Nigella oil on bloodMDA, GSH, SODin normal animals

32. - 50%

33. + 66%

34. + 50%

35. Effect of Nigella oil on bloodMDA, GSH, SODin sensitized animals

36. - 27% - 35%

37. +16% +25%

38. +36%

39. Summary & conclusion Administration of Nigella sativa oil produced the following effects in normal animals andin sensitized animals: • Inhibited the release of histamine • Inhibited the generation of leukotrienes • Increased the release of PGE2 • Inhibited lipid peroxidation • Raised blood glutathione and SOD activity These effects were more marked with the higher dose

40. Conclusions Nigella oil has a prominent anti-inflammatory activity It has antioxidant effects & thus may protect the airways of asthmatic patients against free radical-induced damage These data support and explain the beneficial effect of the oil in ameliorating bronchial asthma and other inflammatory conditions

41. Chemical composition of fixed & volatile oil Thymoquinone (TQ): The major component of the volatile oil; ~ 0.2%w/v in fixed oil Thymohydroquinone: A crystalline substance resulting from refrigeration of the volatile oil Nigellone: The carbonyl polymer of TQ Nigellimine: An isoquinoline alkaloid, found in trace amounts in the seeds Sterols: Cholesterol, Campesterol, Stigmasterol, -sitosterol, -spinasterol

42. Fatty acid composition of the fixed oil of Nigella sativa seeds Linoleic 54.00 - 58.13 % Palmitic 11.97 - 14. 64% Oleic 3.45 - 25.67% Eicosadienoic 2.69 - 2.87% Stearic 2.45 - 2.97% Alpha-linolenic 0.47 - 0.97% Arachidic 0.20 - 0.25% Myristic 0.20 - 0.23

43. Thank you