Nerium oleander`in liyofilize sıvı distilatının rat embriyoları gelişimi üzerine toksik ve teratojen etkilerinin in vitro kültür ortamında araştırılması

Yıl 2014, Cilt: 24 Sayı: 2, 58 - 63, 01.01.2014

Zeliha Fazlıoğulları İsmihan İlknur Uysal Ahmet Levent Baş Nadire Ünver Doğan Ahmet Kağan Karabulut Hasan Acar

Öz

Amaç: Nerium oleander, her mevsimde yeşil kalan bir çalı türüdür. Halk arasında antidiabetik, antilipidemik ve geniş spektrumlu bir ilaç olarak kullanılır. Bu çalışmada Nerium oleander sıvı distilatının kültür ortamında gelişen rat embriyolarına etkilerinin araştırılması planlandı. Gereç ve yöntem: 9,5 günlük rat embriyoları, değişik konsantrasyonlarda 0.2 - 8 mg/ml Nerium oleander ilave edilen rat serumunda 48 saat kültüre edildi. Her bir konsantrasyon için en az 10 embriyo kullanıldı. İlacın sıvı distilatı hidrodistilasyon metodu ile elde edildi. Nerium oleander’in toplam morfolojik skor, yolk sak çapı, tepe-kıç mesafesi ve somit sayısı gibi embriyonik gelişimsel parametreler üzerine doz bağımlı etkileri morfolojik metod ile karşılaştırıldı. Embriyolar malformasyonlar bakımından da değerlendirildi. İlacın genotoksisitesi DNA fragmentasyon metodu ile değerlendirildi. Bulgular: Değişik konsantrasyonlardaki Nerium oleander ilavesi ile kültüre edilen embriyolar kontrol grubu ile karşılaştırıldığında ilacın herhangi bir büyüme geriliği ya da malformasyona sebep olmadığı gözlendi. Sonuç: Bu sonuçlar Nerium oleander’in gebelikte, gelişen embriyo üzerine negatif etkisi olmadığını göstermiştir. Ancak bu veriler ileri in vivo deneyler ve hücre hattı çalışmaları ile desteklenmelidir

Anahtar Kelimeler

Nerium oleander, toksisite, teratojenite, embriyo kültürü

Investigation of developmental toxicity and teratogenicity of lyophylised aqueous distillate of Nerium oleander on rat embryos cultured in vitro

Öz

Objectives: Nerium oleander, an evergreen shrub, is used in folklore medicine as an antidiabetic and antilipidemic and exhibits a wide spectrum of bioactivities. This study was planned to investigate the effects of Nerium osleander lyophilised distillate on developing rat embryos in culture. Materials and methods: Postimplantation rat embryos of day 9.5 were cultured for 48 h in whole rat serum with the addition of various concentrations of Nerium oleander distillate 0.2- 8 mg/ml . At least 10 embryos were used for each con- centration. Aqueous distillate of Nerium oleander was obtained by hydrodistillation method. Dose-dependent effects of Nerium ole- ander on embryonic developmental parameters such as total morphological score, yolk sac diameter, crown-rump length and somite number were compared using morphological method. Embryos were also evaluated for the presence of any malformations. Results: Compared with the controls, the addition of different concentrations of Nerium oleander did not cause any growth retardation and/ or malformations. Conclusions: These results indicate that the use of Nerium oleander in pregnancy may not affect the developing embryos negatively. However, these findings should be supported by further in vivo experiments as well as cell-line studies

Anahtar Kelimeler

Nerium oleander, toxicity, teratogenicity, whole embryo culture

Kaynakça

• Begum S, Siddiqui BS, Sultana R, Zia A, Suria A. Bio-active car- denolides from the leaves of Nerium oleander. Phytochemistry 1999;50:435-8.
• Wang X, Plomley JB, Newman RA, Cisneros A. LC/MS/MS analyses of an oleander extract for cancer treatment. Anal Chem 2000;72:3547-52.
• Ishikawa A, Yamashita H, Hiemori M, et al. Characterization of in- hibitors of postprandial hyperglycemia from the leaves of Nerium indicum. J Nutr Sci Vitaminol (Tokyo) 2007;53:166-73.
• Al-Qura’n S. Ethnopharmacological survey of wild medicinal plants in Showbak, Jordan. J Ethnopharmacol 2009;123:45-50.
• Osterloh J, Herold S, Pond S. Oleander interference in the digoxin radioimmunoassay in a fatal ingestion. JAMA 1982;247:1596-7.
• Adome RO, Gachihi JW, Onegi B, Tamale J, Apio SO. The cardioto- nic effect of the crude ethanolic extract of Nerium oleander in the isolated guinea pig hearts. Afr Health Sci 2003;3:77-82.
• Longford SD, Boor PJ. Oleander toxicity: an examination of hu- man and animal toxic exposures. Toxicology 1996;109:1-13.
• Gayathri V, Ananthi S, Chandronitha C, et al. Cardioprotective effect of Nerium oleander flower against isoproterenol-induced myocardial oxidative stress in experimental rats. J Cardiovasc Pharmacol Ther 2011;16:96-104.
• Heard K. Digoxin and therapeutic cardiac glycosides. In: Dart RC, ed. Medical Toxicology, 3rd ed. Lippincott Williams & Wilkins. Philadelphia, PA 2004;1667.
• Siemens LM, Galey FD, Johnson B. The clinical, cardiac and pat- hophsiological effect of oleander toxicity in horse. J Vet Intern Med 1995;9:217.
• Oryan A, Maham M, Rezakhani A, Maleki M. Morphological tu- dies on experimental oleander poisoning in cattle. Zentralblatt fur Vet Med 1996;43:625-34.
• Adam SE, Al-Yahya MA, Al-Farhan AH. Toxicity of Nerium olean- der and Rhazya stricta in Najdi sheep: hematologic and clinicopat- hologic alterations. Am J Chin Med 2002;30:255-62.
• Aslani MR, Movassaghi AR, Janati-Pirouz H and Karazma M. Ex- perimental oleander (Nerium oleander) poisoning in goats: a clini- cal and pathological study. Iranian Journal of Veterinary Research 2007;8:58-63.
• Barbosa RR, Fontenele-Neto JD, Soto-Blanco B. Toxicity in goats caused by oleander (Nerium oleander). Res Vet Sci 2008;85:279-81.
• New DAT. Whole embryo culture and the study of mammalian embryos during organogenesis. Biol 1978;53:81-122.
• Brown NA, Fabro SE. Quantitation of rat embriyonic development in vitro: a morphological scoring system. Teratology 1981;24:65- 78.
• Van Maele-Fabry G, Delhaise F, Picard JJ. Morphogenesis and quantification of the development of post-implantation mouse embryos. Toxicol In Vitro 1990;4:149-56.
• Kris-Etherton PM, Hecker KD, Bonanome A, Coval SM, Binkoski AE, Hilpert KF, Griel AE, Etherton TD. Bioactive compounds in foods: their role in the prevention of cardiovascular disease and cancer. Am J Med 2002;113:71-88.
• Gayathri V, Ananthi S, Chandronitha C, Sangeetha MK, Vasanthi HR. Hypolipidemic potential of flowers of Nerium oleander in high fat diet-fed Sprague Dawley rats. Nat Prod Res 2011;25:1110-4.
• Bas AL, Demirci S, Yazihan N, Uney K, Ermis Kaya E. Nerium oleander Distillate Improves Fat and Glucose Metabolism in Hi- gh-Fat Diet-Fed Streptozotocin-Induced Diabetic Rats. Int J En- docrinol 2012;2012:947187.
• Al-Yahya MA, AL-Farhan AH, Adam SE. Preliminary toxicity study on the individual and combined effects of Citrullus colo- cynthis and Nerium oleander in rats. Fitoterapia 2000;71:385-91.
• Pathak S, Multani AS, Narayan S, Kumar V, Newman RA. Anvir- zel, an extract of Nerium oleander, induces cell death in human but not murine cancer cells. Anticancer Drugs 2000;11:455-63.
• Bandara V, Weinstein SA, White J, Eddleston M. A review of the natural history, toxinology, diagnosis and clinical management of Nerium oleander (common oleander) and Thevetia peruviana (yel- low oleander) poisoning. Toxicon 2010;56:273-81.
• Al-Farwachi MI, Rhaymah MS, Al- Badrani BA. Acute toxicity of Nerium oleander aqueous leaf extract in rabbits. Iraqi J Vet Sci 2008;22:1-4.
• Szabuniewicz M, Schwartz WL, McCrady JD. Experimental ole- ander poisoning and treatment. Southwestern Vet 1972;25:105-14.
• Thilagar S, Thirumalaikolundusubramanian P, Gopalakrishnan S, akshmikandan R, Ayyappan A, Subramanian R. Possible yellow oleander toxicity in a neonate. Indian Pediatr 1986;23:393.
• Menegola E, Broccia ML, Di Renzo F, Giavini E. Acetaldehyde in vitro exposure and apoptosis: a possible mechanism of teratogene- sis. Alcohol 2001;23:35-9.
• Torchinsky A, Brokhman I, Shepshelovich J, et al. Increased TNF-alpha expression in cultured mouse embryos exposed to te- ratogenic concentrations of glucose. Reproduction 2003;125:527- 34.
• Singh G, Sinha N, Mahipag GS. Role of apoptosis in mediating sa- licylic acid-induced teratogenesis in vitro. Toxicol Mech Methods 2009;19:161-8.
• Singh G, Sinha N. Involvement of apoptosis in mediating mitom- ycin C-induced teratogenesis in vitro. Toxicol Mech Methods 2010;20:190-6.