Ruhi KABAKÇI, Ömer VARIŞLI, Abdulkadir KAYA, İlktan BAŞTAN, Seher ŞİMŞEK

Boğa sperması motilite parametreleri üzerine dietilheksil fitalatın etkisi

Dietilheksil fitalat, sert yapılı plastiklere esneklik katmak amacıyla dünya genelinde yaygın bir şeklide kullanılan, çevresel toksik bir kimyasaldır. İçerisinde bulunduğu ve gevşek bağlarla tutunduğu ürünlerden kolayca ayrılarak çevreye yayılan dietilheksil fitalat sindirim, solunum ve deri teması yollarıyla insan ve hayvanların vücuduna geçmekte ve çeşitli toksik etkilere neden olmaktadır. Bu çalışmanın amacı, dietilheksil fitalatın boğa sperması üzerine in vitro toksik etkisini, bilgisayarlı sperm analiz sistemi ile araştırmaktır. Boğalardan suni vajen ile elde edilen spermalar, fosfat tampon solüsyonu ile 50x106/ml spermatozoa olacak şekilde sulandırıldı ve dimetil sülfoksitte çözdürülen dietilheksil fitalatın 0, 1, 10, 100, 250 ve 500 µg/ml dozlarına maruz bırakılarak 1, 2, 3 ve 4 saat süreyle 37 °C su banyosu içerisinde inkübe edildi. Her inkübasyon süresinin sonunda kontrol ve deneme gruplarından alınan sperma örneklerinin motilite parametreleri analiz edildi. Yapılan değerlendirme sonucunda, dietilheksil fitalatın çalışmada kullanılan düşük dozlarında, doğrudan spermatozoa üzerine toksik etkisi tespit edilemezken, bu etkinin yüksek konsantrasyonlarda zamanla ve doza bağlı olarak ortaya çıktığı belirlendi. Özellikle 500 µg/ml dietilheksil fitalat maruziyetinin sperma hareketlilik parametrelerinden, ortalama yol hızı ve doğrusal hızı 2. saatten itibaren, total motilite ve progresif motiliteyi ise 3. saatten itibaren kontrol gruplarına göre önemli derecede azalttığı belirlendi (p<0.05). İn vitro olarak elde edilen sonuçlar dietilheksil fitalatın erkek üreme organlarında toksik birikimine bağlı olarak boğalarda spermatogenezisi etkiliye bileceğini yönündedir.

Anahtar Kelimeler:

Boğa, CASA, DEHP, Motilite, Sperma

Effect of diethylhexyl phthalate on sperm motility parameters in bull

Diethylhexyl Phthalate is an environmentally toxic chemical commonly used in worldwide to make rigid plastics flexible. Due to its loosely bonded to plastic products, diethylhexyl phthalate easily leaches into the environment. After humans and animals exposure to diethylhexyl phthalate via digestion, respiration and skin contact, it causes various toxic effects in the body. The aim of this study was to investigate the in vitro toxic effect of diethylhexyl phthalate on bull semen motility via a computerized sperm analysis system. Semen collected from the bulls with artificial vagina were diluted with phosphate buffer solution to 50x106/ml spermatozoa and incubated in a 37 ° C water bath for 1, 2, 3 and 4 h by exposing to 0, 1, 10, 100, 250 and 500 µg/ml doses of diethylhexyl phthalate dissolved in dimethyl sulfoxide. At the end of each incubation period, the motility parameters of semen samples taken from control and experimental groups were analyzed. As a result of the evaluation, the toxic effects of diethylhexyl phthalate on the spermatozoa were not determined at low doses used in this study, but this effect occurred at high concentrations by the time. It was observed that exposure of diethylhexyl phthalate particularly at the doses of 500 µg/ml significantly decreased the velocity of average path and linear velocity parameters after 2 h, total motility and progressive motility parameters after 3 h compared to control groups (p<0.05). The in vitro results suggest that toxic accumulation of diethylhexyl phthalate in male reproductive organs can affect fertility in bovine.

Keywords:

Bull, CASA, DEHP, Motility, Sperm,

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Andrade AJ, Grande SW, Talsness CE, Gericke C, Grote K, Golombiewski A, et al. A dose response study following in utero and lactational exposure to di-(2-ethylhexyl) phthalate (DEHP): reproductive effects on adult male offspring rats. Toxicology. 2006;228(1):85-97.
Hannon PR, Flaws JA. The effects of phthalates on the ovary. Front Endocrinol. 2015;6:8.
NTP-CERHR. NTP-CERHR Monograph on the Potential Human Reproductive and Developmental Effects of Di(2-Ethylhexyl) Phthalate (DEHP). NIH Publication; 2006. Report No.: 06-4476 Contract No.: 06-4476.
Ceresana. Plasticizers – Study: Market, Analysis, Trends | Ceresana 2016 [updated 2016. Available from: http://www.ceresana.com/en/market-studies/chemicals/plasticizers/.
ATSDR. ATSDR - ToxFAQs™: Di(2-ethylhexyl)phthalate (DEHP) Agency for Toxic Substances &Diease Registry2002 [updated 2002. Available from: https://www.atsdr.cdc.gov/toxfaqs/TF.asp?id=377&tid=65.
Lorz PM, Towae FK, Enke W, Jäckh R, Bhargava N, Hillesheim W. Phthalic acid and derivatives. Ullmann's encyclopedia of industrial chemistry. 2007.
NTP. Report on Carcinogens. Research Triangle Park; 1998.
Fierens T, Van Holderbeke M, Willems H, De Henauw S, Sioen I. Phthalates in Belgian cow’s milk and the role of feed and other contamination pathways at farm level. Food Chem Toxicol. 2012;50(8):2945-53.
Grossman D, Kalo D, Gendelman M, Roth Z. Effect of di-(2-ethylhexyl) phthalate and mono-(2-ethylhexyl) phthalate on in vitro developmental competence of bovine oocytes. Cell Biol Toxicol. 2012;28(6):383-96.
Hines EP, Calafat AM, Silva MJ, Mendola P, Fenton SE. Concentrations of phthalate metabolites in milk, urine, saliva, and Serum of lactating North Carolina women. Environ Health Perspect. 2009;117(1):86-92.
Kalo D, Hadas R, Furman O, Ben-Ari J, Maor Y, Patterson DG, et al. Carryover Effects of Acute DEHP Exposure on Ovarian Function and Oocyte Developmental Competence in Lactating Cows. PLoS One. 2015;10(7):1-25.
Erkekoglu P, Giray BK, Kızilgün M, Rachidi W, Hininger-Favier I, Roussel A-M, et al. Di (2-ethylhexyl) phthalate-induced renal oxidative stress in rats and protective effect of selenium. Toxıcol Mech Method. 2012;22(6):415-23.
Erkekoglu P, Zeybek ND, Giray BK, Rachidi W, Kızılgün M, Hininger‐Favier I, et al. The effects of di (2‐ethylhexyl) phthalate on rat liver in relation to selenium status. Int J Clin Exp Pathol. 2014;95(1):64-77.
Posnack NG, Swift LM, Kay MW, Lee NH, Sarvazyan N. Phthalate exposure changes the metabolic profile of cardiac muscle cells. Environ Health Perspect. 2012;120(9):1243.
Aung KH, Win-Shwe TT, Kanaya M, Takano H, Tsukahara S. Involvement of hemeoxygenase-1 in di(2-ethylhexyl) phthalate (DEHP)-induced apoptosis of Neuro-2a cells. J Toxicol Sci. 2014;39(2):217-29.
Pant N, Pant A, Shukla M, Mathur N, Gupta Y, Saxena D. Environmental and experimental exposure of phthalate esters: the toxicological consequence on human sperm. Hum Exp Toxicol. 2011;30(6):507-14.
Zhang GL, Zhang XF, Feng YM, Li L, Huynh E, Sun XF, et al. Exposure to bisphenol A results in a decline in mouse spermatogenesis. Reprod Fertil Dev. 2013;25(6):847-59.
Bahrami N, Goudarzi M, Hosseinzadeh A, Sabbagh S, Reiter RJ, Mehrzadi S. Evaluating the protective effects of melatonin on di (2-ethylhexyl) phthalate-induced testicular injury in adult mice. Biomed Pharmacother. 2018;108:515-23.
Głombik K, Basta-Kaim A, Sikora-Polaczek M, Kubera M, Starowicz G, Styrna J. Curcumin influences semen quality parameters and reverses the di (2-ethylhexyl) phthalate (DEHP)-induced testicular damage in mice. J Pharmacological Reports. 2014;66(5):782-7.
Golshan M, Hatef A, Socha M, Milla S, Butts IA, Carnevali O, et al. Di-(2-ethylhexyl)-phthalate disrupts pituitary and testicular hormonal functions to reduce sperm quality in mature goldfish. Aquat Toxicol. 2015;163:16-26.
Kwack SJ, Lee B-M. Comparative cytotoxicity and sperm motility using a computer-aided sperm analysis system (CASA) for isomers of phthalic acid, a common final metabolite of phthalates. J Toxıcol Env Heal A. 2015;78(16):1038-50.
Virtanen H, Rajpert-De Meyts E, Main K, Skakkebaek N, Toppari J. Testicular dysgenesis syndrome and the development and occurrence of male reproductive disorders. Toxicol Appl Pharmacol. 2005;207(2):501-5.
Jarošová A. Phthalic acid esters (PAEs) in the food chain. J Czech J Food Sci. 2006;24:223-31.
Krejčíková M, Jarošová A. Phthalate in cow milk depending on the method of milking. J Medelnet. 2013:592-6.
Pant J, Ranjan P, Deshpande SB. Bisphenol A decreases atrial contractility involving NO-dependent G-cyclase signaling pathway. J Appl Toxicol. 2011;31(7):698-702.
Yurdakok-Dikmen B, Stelletta C, Tekin K, Kuzukiran O, Daskin A, Filazi A. Effects of phthalates on bovine primary testicular culture and spermatozoa. Cytotechnology. 2019:1-13.
Lukácová J, Jambor T, Knazická Z, Tvrdá E, Lukác NJTJoM, Biotechnology, Sciences F. Bıs (2-Ethylhexyl) Phtalate Affects Spermatozoa Motılıty Durıng Short-Term In Vıtro Cultıvatıon. Journal of microbiology, biotechnology and food sciences. 2015;4:73.
Yeste M. Sperm cryopreservation update: Cryodamage, markers, and factors affecting the sperm freezability in pigs. Theriogenology. 2016;85(1):47-64.
Mortimer D, Mortimer ST. Computer-aided sperm analysis (CASA) of sperm motility and hyperactivation. In: Aston DTCI, editor. Spermatogenesis. Methods in Molecular Biology: Springer; 2013. p. 77-87.