67J Contemp Med Sci | Vol. 2, No. 6, Spring 2016: 67–69 Research Objectives The present study was conducted to investigate the effect of Nigella sativa on hormone reproduction and thyroid function in female rats. Methods Twelve females rats were used and divided randomly into two groups (6 animals for each) first group served as treatment and gave Nigella sativa oil (1ml /kg BW/day ) orally according to body weight for a period of 30 consecutive days. While other group drenched with normal saline at same dose that above mentioned and served as control. Results The result revealed significant increase in levels of LH, Estrogen T3 and T4 and significant decrease in level of TSH. Histological sections of thyroid gland revealed presence vaculation in colloid of thyroid follicle. Conclusion In conclusion, the Nigella sativa oil caused the elevation of thyroid hormones as well as LH and estrogen but still within normal value. Keywords Nigella sativa, rat, LH, T3, T4, TSH Study the effect of Nigella Sativa on thyroid function and reproductive hormone of female rat Wafaa Kadhim Jasima, Mayada Sahib Hassana, Ghsoon Ghanem Keamb ISSN 2413-0516 aDepartment of Veterinary physiology and pharmacology, College of Veterinary Medicine, University of Karbala, Karbala, Iraq. bDepartment of Clinical Laboratories, College of Applied Medical Sciences, University of Karbala, Karbala, Iraq. Correspondence to Wafaa Kadhim Jasim (email: wafaa.Gasom@uokerbala.edu.iq ). (Submitted:12 March 2016 – Revised version received: 8 April 2016 – Accepted: 30 April 2016 – Published online: 26 June 2016) Introduction A herbaceous plant, called Nigella sativa (Ns) has different names throughout the world such as black seed, black caraway and black cumin. The black seed oil is reported to be beneficial due to its composition of many components such as aromatic oils trace ele- ments and vitamins.1 Since Nigella sativa oil possesses high ratio of unsaturated fatty acids such as linoloic acid which in turn play an important positive role in function of reproductive system.2 A high percentage of medicine plants have pharmacolog- ical principles, so they are useful as curatives for many ail- ments. According to the reports of World Health Organization (WHO), Traditional medicine won the trust of 70–80% of people in primary health care.3 The plants and their derivatives have a key role in world health. They have long been known to possess biological activity. And from which it draws about 30% of all modern medicines.4 In addition, the plants used for a long time in the folklore medicine to improve fertility by enhancing fertility recipes and aphrodisiacal qualities.5,6 Nigella sativa L. plant belonging to the family called Ranunculaceae,7 which is also called black seed and often their seeds used in folk medicine in some Asian countries and in the Middle East to promote the health and well used in the treat- ment of various diseases.8,9 Several reports confirmed the usefulness of black seed oil because of having more than one hundred of components as vol- atile oil, vitamins and trace elements.1 Recently, clinical and animal studies revealed that the black seed extract has many therapeutic effects. N. sativa seeds have also been used as a nat- ural remedy to promote female menstruation, galactagogue, car- minative, laxative and gastroprotective in traditional medicine.12 Anti-tumor,13,14 anti-anxiety,15 anti-microbial,16,17 anti- parasitic properties,10,11 anti-inflammatory18 and anti-oxidant,19 diuretic and hypotensive, genoprotective, hepato-protective and antidiabetic11 as well as bronchodilator activity and estro- genic activity.20 Despite the progress in N. sativa seeds research in the last decade, the biological and physiological effects of N. sativa seeds still controversial and needs more investigation. So the present study aimed to evaluate N. sativa seeds oil as a supportive traditional medicine on hormones involved in female repro- duction and on thyroid function. Material and Methods Animal and experimental design: The present study was con- ducted on the female rats (Rattus norvegicus) to evaluate the effect of Nigella sativa oil on some hormonal indices of healthy female rats. Twelve female rats were divided randomly into two groups (6 animals for each) first group served as treatment and gavaged Nigella sativa oil (1ml/kg BW/day) orally according to body weight for a period of 30 consecutive days. While other group drenched with normal saline at same dose that above mentioned and served as control group. At the end of the experiment, blood samples were collected under chloro- form anaesthesia via cardiac puncture using sterile disposable syringes at baseline(pre-treatment), day 30 after treatment. The blood samples were then centrifuged at 3000 rpm for 10 minutes to separate the serum. The serum was stored at −80°C until assays were carried out. All tests were performed according to the manufacturer’s instructions.21 Hormonal assay was performed. Estrogen and LH, T3, T4 and TSH levels were measured by using ELISA kit (Bio Merieux). All rats were sacrificed by cervical dislocation under anesthesised and then midline laparotomy was performed. Resected thyroid gland specimens of each rat in all groups were fixed in 10% buffered formaldehyde for 24 hours and his- tological procedure was performed according to Ref. (22 ). Statistical analysis: Two sample Independent t test was used, and the data were expressed as means ± standard devia- tion. The data were analyzed using SPSS windows program version 15.23 Results and Discussion The hormonal activities of LH and estrogen showed significant increase in the treated groups (Table 1) while stayed the same in the control group. 68 J Contemp Med Sci | Vol. 2, No. 6, Spring 2016: 67–69 Study the effect of Nigella Sativa on thyroid function and reproductive hormone of female rat Research Wafaa Kadhim Jasim et al. 6. McDonald A. A Botanical perspective on the identity of Soma (Nelumbo nucifera Gaertn) 2004. Based on Scriptural and Iconographic Records. New York: Botanical Garden Press. 7. D’Cruz SC, Vaithinathan S, Jubendradass R, Mathur PP. Effects of plants and plant products on the testis. Asian J Androl. 2010;12:468–479. 8. Mozaffarian VA. Dictionary of Iranian Plants Names. Tehran: Farhang Moaser Publishers; 1998. p. 365. 9. El-Kadi A, Kandil O. The black seed (N sativa) and immunity- its effects on human T-cell subset. Fed Proc. 1987;46:122–126. 10. Swamy SMK, Tan BKH. Extraction, isolation and characterization of antitumor principle, alpha-hederin from the seeds of Nigella sativa. Plant med. 2001;67:29–32. References 1. Ali BH, Blunden G. Pharmacological and toxicological properties of Nigella sativa. Phytotherapy Res. 2003;17:299–305. 2. Feller SJ, Sauer FD, Kramer JG. Steady-state rates of linoleic acid biohydrogenation by ruminal bacteria in continuous culture. J Dairy Sci. 1995;78:1815. 3. Bashandy AES. Effect of fixed oil of Nigella sativa on male fertility in normal and hyperlipidemic rats. Int J Pharmacol. 2007;3: 27–33. 4. World Health Organization. Traditional Medicine Strategy 2002–2005. Geneva: 2002. 5. Burns MM. Alternative medicine: Herbal preparation. Clin Ped Emerg Med. 2000;1:186–190. In present experiments, the Ns oil treatment led to signif- icant increase in LH levels which may be due to the direct effect of oil on hypothalamus which in turn increases Gonad- otropic Releasing Hormone (GnRH), furthermore fatty acids can stimulate GnRH-dependent pathways that initiate changes in gonads function.24 The positive increased effect of estrogen concentration in treated groups is may be attributed to the contents of the Ns oil especially thymoquinone that enter in building of cholesterol which is important source of cholesterol esters that may have a role in estrogen synthesis.25 Regarding thyroid function, the results of T3, T4 and TSH serum level of control and treatment groups are shown in Table 1, The plasma concentration of T3 and T4 are signifi- cantly (P ≤ 0.005) increased and TSH serum level significantly (P < 0.005) decreased in treatment groups compared with con- trol groups. The treatment with oral administration of NS increased T4 levels in rabbits.26 These results indicated that oral administration of NS lead to hyperthyroidism in mice; likewise, other study reports that the oral administration of NS not only increased serum T3 and decreased TSH but also has an anti-oxidant effect.27 The oral administration with NS could raise T3 level without changing T4 and TSH serum concentration levels.28 Further work is suggested for evaluating the effect of NS on the serum concentration of thyroid hormones to clarify the pos- sible mechanism of action. Histological result of thyroid gland revealed normal appearance of thyroid tissue and intact folli- cular structure in control rats, thyroid follicles lined with cuboidal epithelial lining filled with colloid (Fig. 1). Thyroid gland of female rats treated with Ns showed no histological changes when compare with the control except that some of the follicles were enlarged and containing vacuolated colloid as appear from Figure 2. This might be attributed to increased endocytotic activity of many follicular cells reflecting a com- pensatory mechanism to the augmented release of the stored hormones in the follicular lumen.29 Conclusion In conclusion, the Nigella sativa oil caused elevation of thyroid hormones as well as LH and estrogen but still within normal value and histological sections of thyroid gland revealed pres- ence vaculation in colloid of thyroid follicle.  Table 1. Shows effect of Nigella sativa oil on some hormones in female rats (Mean ± SE) Parameter Groups LH (μIU/ml) Estrogen (pg/ml) Total T3 (ng/dl) Total T4 (μg/dl) TSH (μlU/ml) Control 0.42 ± 0.01 29.08 ± 0.75 0.13 ± 0.02 72.04 ± 1.29 0.05 ± 0.006 Treatment 0.31 ± 0.12* 41.27 ± 1.42* 1.17 ± 0.07* 93.64 ± 1.27* 0.02 ± 0.01* *represent significant difference at (p ≤0.05). Fig. 1 Light micrograph for thyroid gland of control rat, shows normal architecture, thyroid follicles, filled with colloid (star) and lined by cuboidal thyrocytes (thin arrow) (H&E) 100X. Fig. 2 Light micrograph of histological changes in thyroid gland of female rat treated with nigella sativa shows thyroid follicles vary in size (arrow), some of follicles with vacuolated colloid (stars). H&E, 100x. 69J Contemp Med Sci | Vol. 2, No. 6, Spring 2016: 67–69 Research Study the effect of Nigella Sativa on thyroid function and reproductive hormone of female ratWafaa Kadhim Jasim et al. 11. Semsam Shariat H. Medicinal plants. 2nd Ed. Esfahan: Chaharbagh; 2007. p. 32. 12. Zargari A. Medicinal Plants. 6th Ed. Tehran: University Press; 1990. pp. 43–44. 13. 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;11:6662–6666. 14. David RW, Omar AG, Peter AC. The in vitro antitumor activity of some crude and purified components of block seed (Nigella sativa) Anticancer Res. 1998;18:1527–1532. 15. Salomi MJ, Nair SC, Panikkar KR. Inhibitory effects of Nigella sativa and saffron (Crocus sativus) on chemical carcinogenesis in mice. Nutr Cancer. 1991;16:67–72. [PubMed]. 16. Perveen T, Haider S, Kanwal S, Haleem DJ. Repeated administration of Nigella sativa decrease 5-HT Turnover and produces Anxiolytic effects in rats. Pak J Pharm Sci. 2009; 22:139–144. 17. Hanafy MS, Hatem ME. Studies on the antimicrobial activity of Nigella sativa seed (black cumin) J Ethnopharmacol. 1991;34:275–278. 18. Salem ML, Hossain MS. Protective effect of black seed oil from Nigella sativa against murine cytomegalovirus infection. Int J Immunopharmacol. 2000;22:729–740. 19. Houghton PJ, Zarka R, De las Heras B, Hoult JR. Fixed oil of Nigella sativa and derived Thymoquinone inhibit eicosanoid generation in leukocytes and membrane lipid peroxidation. Planta Med. 1995;61:33–36. 20. Burits M, Bucar F. Antioxidant activity of Nigella sativa essential oil. Phytother Res. 2000;14:323–328. 21. Bamosa AO, Ali BA, Sawayan SA. Effect of oral ingestion of Nigella sativa seed on some blood parameters. Saudi Pharm J. 1997;5:126–129. 22. Mescher AL. Junqueira S. Basic Histology Text and Atlas.12th Ed. 2010. p. 1. 23. SPSS Statistical Packages for the Social Sciences. (2001). Statistical software for windows version 13.0 Micrisoft. SPSS, Chicago, IL, USA. 24. Boukhliq R, Martin GB, White CL, Blackberry MA, Murray PJ. Role of glucose, fatty acids and protein in regulating of testicular growth and secretion of gonadotrophin, prolactin, somatotrophin and insulin in the mature ram. J Reprod Fertil. 1997;9:515–524. 25. Wainwarin WLP. The androgen In: Austin CR and Short RV (Eds). Reproduction in mammals. Cambrige Uni Press London; 1979. 26. Sharif SH, Elmahdi BM, Ali Mohammed AM, Mohammed AH. The effects of Nigella sativa L. Ethanolic extract on thyroid function in normal and alloxan- induced diabetic rats. Thyroid Res Pract. 2012;9:48–54. 27. Khalawi AA, Al-Robai AA, Khoja SM, Shaker AS. Can Nigella sativa oil (NSO) reverse hypothyroid status induced by PTU in rat? Biochemical and histological studies. Life Sci J. 2013;10:802–811. 28. Meral I, Yener Z, Ozbek H, Ustun R. Effects of Nigella sativa L. on serum concentrations of thyroid hormones, thyroid stimulating hormone and glucose in alloxan-induced diabetic rabbits. J Vet Med. A Physiology, Pathology, Clinical Medicine. 2001;48:593–599. 29. Saikia UK, Saikia M. Drug-induced thyroid disorders. J Indian Med Assoc. 2006;104(10):585–587.