Setyawati SOEHARTO, I Wayan Arsana WİYASA, Agnes Erna Taulina PURBA, Juneris ARİTONANG, Nurdiana NURDİANA

Sigara Dumanına Maruz Kalan Dişi Sıçanlarda ?-Tocopherol'ün Endometrial Uterin Vaskülarizasyonu, Rahim Serviks Oksidatif Stres ve Proliferasyon Üzerine Etkisi

Amaç: Bu çalışma ?-tocopherol uygulamasının uterin endometrium vaskülarizasyonu arttırmada, uterin servik soksidatif stresinin hibe edilmesi ve sigara dumanına maruz kalan dişi sıçanlarda servikal çoğalmanın artırılabileceğinin mümkün olup olmadığını öğrenmeyi amaçlamıştır. Materyal ve Metod: Yirmibeş adet, 8 haftalık dişi Wistar sıçanı (Rattusnorvegicus) kontrol grubu (maruz bırakılmamış grup) dahil 5 gruba (n=5) ayrılmıştır; sigara dumanına (CS) maruz bırakılmış sıçanlar; sigara dumanına maruz bırakılmış ve 100mg/kg/gün dozluk (CS+AT1) ?-tocopherol'aile muamele edilmiş sıçanlar; sigara dumanına maruz bırakılmış ve 200mg/kg/gündozluk (CS+AT2) ?-tocopherolile e muamele edilmiş sıçanlar; sigara dumanına maruz bırakılmış ve 400mg/kg/gündozluk (CS+AT3) ?-tocopherolile e muamele edilmiş sıçanlar. Endometrial VEGF'nin ekspresyonu, anti-VEGF, primer antikor vebiyotinlenmiş genel sekonderantikor kullanılarak immunohistokimyasal boyama ile gözlenmiştir. MDA düzeyleri kolorimetrik olarak ölçülmüştür. Spiral arterlerin sayısı ve Rahim serviks epitel hücrelerin çoğalması hematoksiline ozin boyalarla histolojik olarak sayılmıştır. Bu araştırma Endonezya Malang'daki (No 477/EC/KEPK-S2-KB/10/2013) Brawijaya Üniversitesi Tıp Fakültesi'nin araştırma etik komitesi tarafından onaylanmıştır. Bulgular: Sigara dumanı control grubuyla karşılaştırıldığında özellikle VEGF seviyesini, spiral arterlerin ve uterin serviks epitel hücrelerinin sayısını düşürmüştür (P0.05).a-tocopherol'ün İkinci ve üçüncü dozları CS grubuna kıyasla (P0.05). Sıçanlarda sigara dumanına maruz kalmanın serviks MDA düzeyini control grubuna(P

The Effects of ?-Tocopherol on Endometrial Uterine Vascularization, Uterine Cervix Oxidative Stress and Proliferation in Female Rats Exposed to Tobacco Smoke

Purpose: This study aimed toknow whether administration of ?-tocopherolable to increase uterine endometrial vascularization, inhibit uterine cervix oxidative stress, and augment cervical proliferation in female rats exposed to tobacco smoke. Material and Methods:Twenty five, 8 weeks old female Wistar rats (Rattusnorvegicus) were divided into 5 groups (n=5 each), including control group (non exposure group); the rats were exposed to tobacco smoke (CS); the rats were exposed to tobacco smoke and treated with ?-tocopherol at dose of 100 mg/kg/day (CS + AT1); rats were exposed to tobacco smoke and treated with ?-tocopherol at dose of 200mg/kg/day (CS + AT2); rats were exposed to tobacco smoke and treated with ?-tocopherol at dose of 400 mg/kg/day (CS + AT3). The expression of endometrial VEGF was observed by immunohistochemical staining using anti-VEGF primary antibody and biotinylated universal secondary antibody. MDA levels were measured colorimetrically. The number of spiralis arterioles and proliferation of uterine cervix epithelial cells were counted histologically withhematoxilin eosin staining. This research has been approved by research ethics committee Faculty of Medicine University of Brawijaya, Malang, Indonesia (Number 477/EC/KEPK-S2KB/10/2013). Results: Tobacco smoke significantly lowered VEGF level, the number of spiralis arterioles, anduterine cervix epithelial cells compared with control group (P < 0.05). Administration of ?-tocopherol in different doses significantly increasethe VEGF level to reachs the level in the control group (P > 0.05). Second and third doses of ?-tocopherolsignificantly increased the number of spiralis arterioles compared with CS group (P < 0.05), to reached the similar level in the control group (P > 0.05).Exposure to tobacco smoke in rats can increase uterine cervix MDA level significantly compared to the control (P< 0.05). The ?-tocopherol in various doses cansignificantly reduce MDA level compared to the CS (P 0.05). Conclusion:Administration of ?-tocopherolcan increase the vascularization in endometrial uterine of rats exposed to tobacco smoke. Besides, ?-tocopherolinhibit uterine cervix oxidative stress, but can't induces the proliferation of uterine cervix epithelial cells.

PDF

___

1. Dechanet C, Anahory T, Daude MJC, Quantin X, Reyftmann L, Hammah S, Hedon B, Dechaud H. Effect of cigarette smoking on reproduction. HumReprod Updates. 2011;17:76-95.
2. Gannon AM, Sta¨mpfli MR, FosterWG. Cigarette smoke exposure leads to follicle loss via an alternative ovarian cell death pathway in a mouse model. ToxicolSci 2012;125:274-84.
3. Andersen MR, Uldbjerg N, Stender S, Sandager P, Aalkjaer C. Maternal smoking and impaired endothelium-depedent nitric oxide-mediated relaxation of uterine small arteries in vitro. Am J ObstetGynecol 2011;204:177.e1-177.e7.
4. Dechanet C, Brunet C, Anahory T, Hamamah S, Hedon B, Dechaud H. Effects of cigarette smoking on embryo implantation and placentation and analysis of factors interfering with cigarette smoke effects (PartII). GynecolObstetFertil. 2011;39:567-74.
5. Bardak Y, Ozerturk Y, Ozguner F, Durmus M, Delibas N. Effect of melatonin against oxidative stress in ultraviolet-B exposed rat lens. Current Eye Res. 2003;20:225-30.
6. Kazi AA, KoosRD. Estrogen-induced activation of hypoxia-inducible factor-1?, vascular endothelial growth factor expression, and edema in the uterus are mediated by the phosphatidylinositol 3-kinase/akt pathway.Endocrinology 2007;148:2363-74.
7. Fonseca-Moutinho JA. Smoking and cervicalcancer. ISRN ObstetGynaecol 2011; 2011:Article ID 847684, 6 pages.
8. Palan PR, Woodall AL, Anderson PS, Mikhail MS. Alpha-tocopherol and alpha-tocopherylquinone levels in cervical intraepithelial neoplasia and cervical cancer. Am J Obstet Gynecol. 2004;190:1407-10.
9. Cicel N, Eryilmaz O.G, Sarikaya E, Gulerman C, YaseminG. Vitamin E effect on controled ovarian stimulation of unexplained infertile women. J Assist Reprod Genet. 2012;29:325-8.
10. Tarin JJ, Perez-Albala S, Pertusa JF, Cano A. Oral administration of pharmacological doses of Vitamins C and E reduces reproductive fitness and impairs the ovarian and uterine functions of female mice. Theriogenology. 2002;57:1539-50.
11. Duran M, Ustunyurt E, Kosus A, Kosus N, Turhan N, Hizli D, Sarac GN, Erdogan D. Does vitamin E prevent tubal damage caused by smoking? A light microscopy and animal study. Eur J ObstetGynecolReprodBiol 2014; http: //dx.doi.org/10/1016/j/ejogrb.2014.01.020.
12. Tome AR, Ferreira PMP, Freitas RM. Inhibitory Action of antioxidants(ascorbic acid or ?-tocopherol) on seizures and brain damage induced by pilocorpine in rats. ArqNeuropsiquiatr 2010;68:355-61.
13. Fraser HM, Wilson H, Silvestri A, Morris DK, Wiegand SJ. The role of vascular endothelial growth factor and estradiol in regulation of endometria angiogenesis and cell proliferation in the marmoset. Endocrinology. 2008;149:4413-20.
14. Gazdar AF. Environmental tobacco smoke, carcinogenesis, and angiogenesis: A double whammy. Cancer Cell. 2003;4:159-60.
15. Venditti CC, Casselman R, Murphy MS, AdamsinSl, Sled JG, Smith GN. Chronic carbon monoxide inhalation during pregnancy augments uterine artery blood flow and uteroplacental vascular growth in mice. Am J PhysiolRegulIntegr Comp Physiol. 2013;305:R939-48.
16. Lee J, Cooke JP. Nicotine and pathological angiogenesis. Life Sci. 2012;91:1058-64.
17. Motejlek K, Palluch F, Neulen J, Grümmer R. Smoking impairs angiogenesis during maturation of human oocytes. Fertility and Sterility. 2006;86:186- 91.
18. Burton GW, Traber MG. Vitamin E: antioxidant activity, biokinetics, and bioavailability. Ann Rev of Nutr.1990;10:357-82.
19. Ozer NK, Sirikci O, Taha S, San T, Moser U, Azzi A. Effect of vitamin E and probucol on dietary cholesterol-induced atherosclerosis in rabbits. Free RadicBiol Med. 1998;24:226-33.
20. Keaney JF Jr, Gaziano JM, Xu A, Frei B, CurranCelentano J, Shwaery GT, Loscalzo J, Vita JA.Lowdose ?-tocopherol improves and high-dose ?- tocopherol worsens endothelial vasodilator function in cholesterol- fed rabbits. J Clin Invest. 1994;93:844- 51.
21. Kontush A, Finckh B, Karten B, Kohlschutter A, Beisiegel U.Antioxidant and prooxidant activity of ?- tocopherol in human plasma and low density lipoprotein. J Lipid Res. 1996;37:1436-48.
22. Versari D, Daghini E, Rodriguez-Porcel M, Sattler K, Galili O, Pilarczyk K, Napoli C, Lerman LO, Lerman A.Chronic antioxidant supplementation impairs coronary endothelial function and myocardial perfusion in normal pigs. Hypertension.2006;47:475- 81.
23. Zhang B, Tanaka J, Yang L, Yang L, Sakanaka M, Hata R, Maeda N, Mitsuda N. Protective effect of vitamin E against focal brain ischemia and neuronal death through induction of target genes of hypoxiainducible factor-1. Neurosci. 2004;126:433- 40.
24. Yao JS, Zhai W, Fan Y, Lawton MT, Barbaro NM, Young WL, Yang GY. Interleukin-6 upregulates expression of KDR and stimulates proliferation of human cerebrovascular smooth muscle cells. J Cereb Blood Flow Metab. 2007;27:510-20.
25. Yao JS, Zhai W, Young WL, Yang GY. Interleukin-6 triggers human cerebral endothelial cells proliferation and migration: the role for KDR and MMP-9. BiochemBiophys Res Commun. 2006;342:1396-404.
26. Ejaz S, Lim CW. Toxicological overview of cigarette smoking on angiogenesis. Environ ToxicolPharmacol. 2005;20:335-44.
27. Moller B, Lindblom B, Olvsson M. Expression of the vascular endothelial growth factor B and C and their receptors in human endometrium during the menstrual cycle. ActaObstetGynecol Scand. 2002;81:817-24.
28. De S, Sengupta A, Chakraborty RN, Das S. Influence of alpha tocopherol during carcinogenesis in uterine cervix of mice. Nutr Res. 2000;20:261-72.