Nalan KAYA TEKTEMUR, Leyla CANPOLAT KOYUTÜRK, Ebru GÖKALP ÖZKORKMAZ, Eda YILDIZHAN, Murat AKKUŞ, Fırat AŞIR

EXAMINATION OF APOPTOTIC CHANGES IN AROMATIC HYDROCARBON BENZENEADMINISTERED RAT TESTICULAR TISSUE WITH HISTOPATHOLOGICAL METHODS

Benzene is an aromatic hydrocarbon and a colorless and odorous liquid. It is used in the production of plastics, detergents, pesticides, and other chemicals. Serious deterioration is seen in sperm production of men who are exposed to hydrocarbons such as ethylbenzene, toluene, xylene, andaromatic solvents found in paints, lacquers, adhesive-like substances in their professions. The purpose of this study was to examine the possible effects of benzene on testicular tissues via histopathological and TUNEL staining methods. In this study, healthy 18 Wistar albino male rats, with a mean age of 8-10 weeks weighing 250-300 g were used. Rats were divided into 3 groups; Group I was the controlgroup. Group II was administered 1 ml/kg benzene for 9 days and Group III 1.5 ml/kg benzene for 5 days via orogastric tube. At the end of the experiment, all rats were decapitated, and their testicular tissues were excised. Stained with Haematoxylin and Eosin, Johnsen scoring was performed for each group. Apoptosis was demonstrated with TUNEL method, marked with the Image J program and statistical analysis was performed. Benzene exposed rat testicular tissues depicted thinning of seminiferous tubule epithelium cytoplasmic vacuoles, affusion of seminiferous tubule epithelium to the lumen, and affusion in spermatogenic cells (desquamation), degenerative changes in germ cells (spermatogonia), degenerative tubule structures, disorganized interstitial tissues, and absence of epithelial integrity were observed. Compared with the control group, the rats exposed to benzene revealed a significant increase in apoptotic cells in the seminiferous tubules via TUNEL staining. It was thought that exposure to benzene resulted in degeneration and increased apoptotic cells in the testicular tissues.

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[1] Adel-Shafy, H.I., Mansour, M.S.M., “A review on polycyclic aromatic hydrocarbons: Source, environmental impact, effect on human health and remediation”, Egyptian Journal of Petroleum, 25, 107-123, 2016. Doi:10.1016/j.ejpe.2015.03.011.
[2] Rengarajan, T., Rajendran, P., Nandakumar, N., Lokeshkumar, B., Rajendran B., Nishigaki, I., “Exposure to polycyclic aromatic hydrocarbons with special focus on cancer”, Asian Pacific Journal of Tropical Biomedicine, 5(3), 182-189, 2015. Doi: 10.1016/S2221-1691(15)30003-84.
[3] Cherrie, J., “Risks of Exposure to Hazardous Substances: Toxicology and Beyond, About Chemical Substances in the Workplace”, European Agency for Safety and Health at Work presentation–EU-OSHA, 2009.
[4] Vincent, R., “CMR chemical substance inventory used in France in 2005 INRS - National Research and Safety Institute, Occupational Health, and Safety, Workbooks– 4th quartile”, 2006.
[5] OSHwiki –EU-OSHA. Occupational safety and health (OSH) homepage on the Internet Web site, Access date: 18/11/2020, Available from: https://oshwiki.eu/wiki/Category:Carcinogenic,_mutagenic,_reprotoxic_(CMR)_substances
[6] Li, K., Jing, Y., Yang, C., Liu, S., Zhao, Y., He, X., et al., "Increased leukemia-associated gene expression in benzene-exposed workers”, Scientific Reports, 4, Article number: 5369, 2014. doi: 10.1038/srep05369.
[7] Kamal, A., Malik, R.N., “Hematological Evidence of Occupational Exposure to Chemicals and Other Factors among Auto-Repair Workers in Rawalpindi, Pakistan”, Osong Public Health and Research Perspectives, 3(4), 29-238, 2012. Doi: 10.1016/j.phrp.2012.10.003.
[8] AFSSET - Ulusal Çevre Sağlığı ve Gıda Güvenliği Ajansı, Üreme Ve Gelişme Özellikli Toksik Madde Listesinin Belirlenmesi Ve Toksikolojik Referans Değerlerinin Analizi Için Bir Sıralama Yönteminin Önerilmesi ,'VTR reprotoksik' uzmanları, 58, 2006. Access date: 18/11/2020, Available from: https://oshwiki.eu/wiki/Category:Carcinogenic,_mutagenic,_reprotoxic_(CMR)_substances
[9] Evans, T.J. “Endocrine disrupters”, Gupta, R.C., (Ed.), Reproductive and Developmental Toxicity, Elsevier Inc. 874-875, 2011.
[10] Beardsley A, O’ Donnell L. “Characterization of normal spermiation and spermiation failure induced by hormone suppression in adult rats”, Biology of Reproduction, 68, 1299-1307, 2003. Doi: 10.1095/biolreprod.102.009811.
[11] Aladakatti, R.H., Nazeer A.R., “Changes in Sertoli cells induced by Azadirachta indica A. Juss leaves in albino rats”, Journal of Basic and Clinical Physiology and Pharmacology, 16, 67-80, 2005. Doi: 10.1515/jbcpp.2005.16.1.67.
[12] Cho, S.I., Damokosh, A.I., Ryan, L.M., Chen, D., Hu, Y.A., Smith, T.J., Christiani, D.C., Xu X., “Effects of exposure to organic solvents on menstrual cycle length”, Journal of Occupational and Environmental Medicine, 43(6), 567-75, 2001. doi: 10.1097/00043764- 200106000-00012.
[13] Rogene, F.H., “Species Differences in the Metabolism of Benzene”, Environmental Health Perspectives, 106 (6), 1173-1175, 1996. Doi: 10.1289/ehp.961041173.
[14] Ganousis, L.G., Goon, T., Zyglewska, K.K., Ross, W.D., “Metabolism in mouse Bone Marrow Stroma: Study of activation and detoxification of benzene metabolites”, Molecular Pharmacology, 42 (6), 1118-1125, 1992. PMID: 1480134.
[15] Nemes, Z. JR., Friis, R.R., Aeschlimann, D., Saurer, S., Paulsson, M, Fesüs, L., “Expression and activation of tissue transglutaminase in apoptotic cells of involuting rodent mammary tissue”, European Journal of Cell Biology, 70 (2), 125-33, 1996. PMID: 8793384.
[16] Zeiss, C.J., “The Apoptosis- Necrosis Continuum: Insights from Genetically Altered Mice” Veterinary pathology, 40, 481-495, 2003. Doi: 10.1354/vp.40-5-481.
[17] White, E., “Death-defying acts: a meeting review on apoptosis” Genes and Development, 7, 2277- 2284, 1993. Doi: 10.1101/gad.7.12a.2277.
[18] Sharma, R., Bhat, R., Goyal, A., Bhardwaj, J.K., “Germ Cells Apoptosis during Spermatogenesis in Mammals”, Journal of Entomology and Zoology Studies, 3(3), 506-515, 2015. ISSN 2320-7078.
[19] Mahdi, A., Rajender, S., Shukla, K., “Apoptosis, spermatogenesis and male infertility” Frontiers in Bioscience, 1(4), 746-754, 2012. Doi: 10.2741/415.
[20] National Research Council. Guide for the Care and Use of Laboratory Animals: Eighth Edition. Washington, DC: The National Academies Press. 2011. Doi: 10.17226/12910.
[21] Johnsen, S.G., “Testicular biopsy score count--a method for registration of spermatogenesis in human testes: normal values and results in 335 hypogonadal males” Hormones, 1(1), 2-25, 1970. Doi: 10.1159/000178170.
[22] Joshi, D.R., Adhikari, N., “An Overview on Common Organic Solvents and Their Toxicity”, Journal of Pharmaceutical Research International, 28(3), 1-18, 2019. Doi: 10.9734/jpri/2019/v28i330203.
[23] Cornish, H.H., Ryan, R.C. Metabolism of benzene in nonfasted, fasted, and aryl-hydroxylase inhibited rats. Toxicology and applied pharmacology, 7(6), 767-71, 1965. Doi: 10.1016/0041- 008x (65)90001-3.
[24] Withey, R.J., Hall, J.W. The joint toxic action of perchloroethylene with benzene or toluene in rats. Toxicology, 4 (1), 5-15, 1975. Doi: 10.1016/0300-483X (75)90017-7.
[25] Hsieh, G.C., Sharma, R.P., Parker, R.D.R. Subclinical effects of groundwater contaminants I: Alteration of and cellular immunity by benzene in CD-1 mice. Archives of Environmental Contamination and Toxicology, 17, 151–158, 1988. Doi: 10.1007/BF01056019.
[26] Singh, R.K., Bansode, F.W., “Benzene- induced histopathological changes and germ cell population dynamics in testes of Sprague Dawley rats”, Journal of Environmental Biology, 32, 687- 694, 2011. ISSN: 02548704.
[27] Ravindranath, H., Aladakatti M.A., Nazeer A. R., Mukhtar A. G. Ghodesawar, “Effect of benzene leaf extract of Ocimum sanctum on testis and spermatogenic pattern in albino rats”, International Journal of Current Research, 5, 22-29, 2010. ISSN: 0975-833X.
[28] Linder, R.E., Hess, R.A., Strader, L.F., “Testicular toxicity and infertility in male rats treated with 1,3- dinitrobenzene”, Journal of toxicology and environmental health, 19(4), 477- 489, 1986. Doi: 10.1080/15287398609530946.
[29] Cárdenas-Valencia, I., Nieto, J., Gasco, M., Gonzales, C., Rubio, J., Portella, J., Gonzales, G.F., “Tropaeolum tuberosum (Mashua) reduces testicular function: effect of different treatment times. Andrologia, 40(6), 352-7, 2008. doi: 10.1111/j.1439-0272.2008.00868.x. PMID: 19032684.
[30] National Toxicology Program. “Toxicology and carcinogenesis studies of divinylbenzene- HP (Cas No. 1321- 74- 0) in F344/ N rats and B6C3F1 mice (inhalation studies)”, National Toxicology Program technical report series, 18 (4), 9- 285, 2006. PMID: 17342197.
[31] Shi, Y., Song, Y., Wang, Y., Wang, Y., Liang, X., Hu, Y., Yu, H., Guan, X., Cheng, J., Yang, K. β-“Benzene hexachloride induces apoptosis of rat Sertoli cells through generation of reactive oxygen species and activation of JNKs and FasL”, Environmental toxicology, 26(2), 124–135, 2011. Doi:10.1002/tox.20536.