International Journal of PharmTech Research CODEN (USA): IJPRIF, ISSN: , ISSN(Online): Vol.9, No.11, pp , 2016

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International Journal of PharmTech Research CODEN (USA): IJPRIF, ISSN: 0974-4304, ISSN(Online): 2455-9563 Vol.9, No.11, pp 246-251, 2016 The Effects of Yasmin on the Histology and Histochemistry of Liver and Kidney in Rabbits Fakhir M. Alzubaidy 1 *, Rafah S. Almuttairi and Aymen A. Bash 2 1 University of Babylon/ College of Pharmacy, Dept. of Laboratory & Clinical Sciences, Iraq. 2 University of Babylon/ College of Pharmacy, Dept. of Pharmaceuticals, Iraq. Abstract : Objective: The combination birth control pill of ethinyl estradiol with drospirenone (YASMIN) is an effective contraceptive which used by a very large number of women worldwide, and this study was designed to investigate the probable histological and histochemical changes of the liver and kidney accompanied with the treatment by yasmin. Methods: 12 females adult rabbits were used in this experiment (6 treated animals and 6 as control group). The treatment by the yasmin contraceptive pills based on the same way as used for women, regarding the body mass index for dosage, then frozen and ordinary tissue sectioning were performed. Results: Histological and histochemical (for alkaline phosphatase ALP- activity) study of both liver and kidney was normal. Conclusion: Yasmin is safe with regard to the histology of the liver and kidney, and the activity of ALP in these tissues. Keywords: Yasmin, Histology and Histochemistry, Liver and Kidney, Rabbits. Introduction Combined oral contraceptives (COCs) represent one of the most commonly prescribed contraceptive methods and are used by more than 100 million women worldwide 1.These pills contain an oestrogen component (ethinyloestradiol, mestranol, oestradiol or its pro-drug oestradiolvalerate) and a progestogen (levonorgestrel, norethisterone, gestodene, desogestrel, drospirenone, nomegestrol, dienogest or cyproterone) 2. Yasmin is a combined oral contraceptive tablet containing the synthetic progestogen, drospirenone and the synthetic oestrogen, ethinyloestradiol 3.A large number of publications refer to the rapid evolution of this contraceptive method, particularly with respect to the reduction in the dose of estrogen and the synthesis of new progestogens. More recently, new contraceptive regimens, specifically those involving continuous or extended use aimed at minimizing the hormone-free interval between one package of pills and another, have been considered both by physicians and their patients 4-6. The combined oral contraceptive pill is an effective contraceptive method which can also offer other benefits. Women rapidly adopted the pill as it allowed the reliable separation of sex and reproduction and gave them the opportunity to plan when to have children. Since then the pill has been further developed to ensure good efficacy while minimizing the adverse effects 2. Also, many studies were showed many observations about the COCs, Long-term cohort studies show that, compared to non-users of the combined oral contraceptive pill, users have lower rates of death from any cause. They also have significantly lower rates of death from cancer, cardiovascular disease and other diseases 7. Another study have evaluated the effects of the continuous use of contraceptive pills on metabolic parameters and coagulation 8.

Fakhir M. Alzubaidy et al /International Journal of PharmTech Research, 2016,9(11): 246-251. 247 Animals and methods: Twelve females adult rabbits were used in this experiment (6 treated animals and 6 as control group). The treatment by the yasmin contraceptive pills based on the same way as used for women (timing and sequence), regarding the body mass index for dosage. Histological examination: Ordinary histological sections of paraffin-embedded liver and kidney (5µ thick), stained with hematoxylin-eosin were performed according to Bancroft and Steven,1982 9. Detection of Alp activity: Frozen sections was prepared for the study of Alp activity in the liver and kidney. Tissues froze immediately after sacrificing the animals, then sectioning was done by freezing microtome. Finally, sections treated with the chemical solutions and stained with methyl green for the detection of enzyme activity. Gomori-Calcium method, 1952 (modified) was depended to detect the activity of Alp, as it considered one of the most important depended methods in histochemical preparations 9. Results and discussion : Histological study : Result showed normal histology of liver and kidney of the treated animals in compare with control group, as shown in the figures (1,2, 3, and 4).These results may reflect the safety of yasmin pills, as previous studies referred to the wide use and safety of this drug 2,3. in spite of the presence of biochemical changes that related to the liver and kidney 10,11,12 Alkaline phosphatase (Alp) Activity : In spite of the risks and undesired physiological and biochemical changes that revealed by many previous studies 10, 13-17, but our histochemical study of Alp activity showed a normal activity in both liver and kidney of both study groups in compare with the normal group as shown in the figures (5, 6, 7, and 8). Again this may related to the safe part of the yasmin, or it may be related to the quantity of the biochemical changes. For example studies referred to reduction in total cholesterol, low-density lipoprotein, and increase in highdensity lipoprotein 10,12. Figure 1 : Histological section in liver of treated animals showing normal histology.

Fakhir M. Alzubaidy et al /International Journal of PharmTech Research, 2016,9(11): 246-251. 248 Figure 2 : Histological section in liver of normal animals (control). Figure 3: Histological section in kidney of treated animals showing normal histology. Figure 4 : Histological section in kidney of normal animals (control).

Fakhir M. Alzubaidy et al /International Journal of PharmTech Research, 2016,9(11): 246-251. 249 Figure 5 :Frozen histological section in liver of treated animal stained with methyl green, showing normal Alp activity (brown color), hepatocytes stained green. Figure 6 :Frozen histological section in liver of normal animal (control), stained with methyl green, showing normal Alp activity (brown color), hepatocytes stained green. Figure 7 :Frozen histological section in kidney of treated animal stained with methyl green, showing normal Alp activity (brown color).

Fakhir M. Alzubaidy et al /International Journal of PharmTech Research, 2016,9(11): 246-251. 250 Figure 8 :Frozen histological section in kidney of normal animal stained with methyl green, showing normal Alp activity (brown color). References: 1. Fu, H. ;Darroch, J. E. ; Haas, T. ;Ranjit, N. (1999). Contraceptive failure rates: new estimates from the 1995 National Survey of Family Growth. FamPlannPerspect31:56 63. 2. Stewart, M. and Black, K. 2015. Choosing a combined oral contraceptive pill. AustPrescr38 (1): 6-11. 3. Bayer HealthCare Pharmaceuticals. (2011).Drospirenone-Containing Combination Oral Contraceptives Briefing Document.Bayer HealthCare Pharmaceuticals Inc. P: 1-137 4. Sulak, P. J. ; Buckley, T. Kuehl, T. J. (2006). Attitudes and prescribing preferences of health care professionals in the United States regarding use ofextended-cycle oral contraceptives. Contraception; 73:41 5. 5. Wiegratz, I. ;Hommel, H. H. ; Zimmermann, T. ;Kuhl, H. (2004).Attitude of German women and gynecologists towards long-cycle treatment with oral contraceptives. Contraception;69:37 42. 6. Machado, B. R. ; de Melo, N. R. ; Jr, M. H. and Cruz, M. A. (2010). Effect of a continuous regimen of contraceptive combination of ethinylestradiol and drospirenone on lipid, carbohydrate and coagulation profiles 7. Hannaford, P. C. ; Iversen, L. ; Macfarlane, T. V. ; Elliott, A. M. ; Angus, V. ; Lee, A. J. (2010). Mortality among contraceptive pill users: cohort evidence from Royal College of General Practitioners Oral Contraception Study. BMJ;340:c927. 8. Machado, R. B. ;Fabrini, P.; Cruz, A. M.; Maia, E. ; da Cunha Bastos, A. (2004). Clinical and metabolic aspects of the continuous use of a contraceptive association of ethinyl estradiol (30 microg) and gestodene (75 microg). Contraception;70:365 70. 9. Bancroft, D. J. and Stevens, A. (1982). Theory and Practice of HistologicalTechniques.2 nd Edition. Chrchill Livingstone. Medical Division of Longman Group Limitted. 10. Taneepanichskul S, Phupong V. (2007). Influence of a new oral contraceptive with drospirenone on lipid metabolism. GynecolEndocrinol;23:347 50. 11. Endrikat J, Klipping C, Cronin M. (2002). An open label, comparativestudy of the effects of a dosereduced oral contraceptive containing 20mcg ethinyl estradiol and 100 mcg levonorgestrel on hemostatic,lipids, and carbohydrate metabolism variables. Contraception;65:215 21. 12. Petitti DB. Clinical practice. (2003). Combination estrogen-progestin oralcontraceptives. N Engl J Med;349:1443 50. 13. Farley TM, Meirik O, Collins J. (1999). Cardiovascular disease and combined oral contraceptives: reviewing the evidence and balancing the risks. Hum ReprodUpdate;5:721-35. 14. Plu-Bureau, G. ;Hugon-Rodin, J. ;Maitrot-Mantelet, L. ;Canonico, M. (2013). Hormonal contraceptives and arterial disease: an epidemiological update. Best Pract Res ClinEndocrinolMetab;27:35-45.

Fakhir M. Alzubaidy et al /International Journal of PharmTech Research, 2016,9(11): 246-251. 251 15. Wu, C. ; Grandi, S. M. ; Filion, K. B. ; Abenhaim, H. A. ; Joseph, L. and Eisenberg, M. J. (2013). Drospirenone-containing oral contraceptive pills and the risk of venous and arterial thrombosis: a systematic review. International Journal of Obstetrics and Gynaecology 120:801 811. *****