SU ARACILIĞIYLA SODYUM FLORÜR KONSANTRASYONLARINA MARUZ KALMA LEYDİG HÜCRELERİNDE SİTOTOKSİSİTE VE OKSİDATİF HASARA NEDEN OLABİLİR

Year 2017, Volume: 18 Issue: 2, 115 - 122, 22.09.2017

Yasemin Aydin

https://doi.org/10.23902/trkjnat.303729

Abstract

Bu çalışma sodyum florürün
(NaF) TM3 Leydig hücrelerinde in vitro’da
sitotoksisite ve oksidatif stres üzerindeki etkilerini araştırmıştır. Sitotoksisite
hücre canlılık ve çoğalma hızı ile ve laktat dehidrogenaz testi ile
belirlenmiştir. Oksidatif hasar, artan lipid peroksidasyon ürünü, reaktif
oksijen türleri (ROS) ve azalan hücresel enzimatik ve enzimatik olmayan
antioksidanlar yoluyla ölçülmüştür. Leydig hücreleri, Türkiye'de içme suyunun
günlük alımına dayanan en düşük ve en yüksek florür konsantrasyonları olan iki
farklı NaF (2ppm ve 16ppm) konsantrasyonuna 24 saat, 48 saat ve 72 saat maruz
bırakılmıştır. Bu çalışma, NaF'ye maruz bırakılan Leydig hücrelerinde,
konsantrasyon ve zamana bağlı olarak, hücre canlılığı ve proliferasyonundaki
azalmanın ve oksidatif stresin arttığını göstermiştir. Bu sonuçlar florür
uygulanmasının Leydig hücrelerinde ve özellikle hücrelerin antioksidan
sisteminde hasara neden olabileceğini önerir. Bu aynı zamanda oksidatif stresin
Leydig hücresi fonksiyonu üzerinde bozulmalarda önemli bir rol oynadığını
kanıtlamaktadır. 

Keywords

Florür, sitotoksisite, lipid peroksidasyonu, ROS, antioksidan sistem, Leydig hücreleri

EXPOSURE TO SODIUM FLUORIDE VIA DRINKING WATER CAUSE CYTOTOXICITY AND OXIDATIVE DAMAGE IN LEYDIG CELLS

Abstract

This study investigated the in vitro effects of sodium fluoride (NaF) on cytotoxicity and
oxidative stress in TM3 Leydig cells. Cytotoxicity was determined with cell
viability and proliferation rate, and lactate dehydrogenase leakage assay.
Oxidative damage was measured by increasing lipid peroxidation product,
reactive oxygen species (ROS) and decreasing cellular enzymatic and
non-enzymatic antioxidants. Leydig cells were exposed to two different
concentrations of NaF (2ppm and 16ppm) for 24h, 48h and 72h which are
considered to be the lowest and highest fluoride concentrations based on daily
intake of drinking water in Turkey. The present study showed that cell
viability and proliferation decreased and oxidative stress increased in
NaF-exposed Leydig cells in a concentration- and time-dependent manner. The
findings suggest that fluoride-treatment can cause damage in Leydig cells,
especially in antioxidant system of the cells. The results also proved that
oxidative stress plays an important role in impairments on Leydig cell function.

Keywords

Fluoride, cytotoxicity, lipid peroxidation, antioxidant system, Leydig cell, reactive oxygen species

References

• 1. Akdoğan, M., Bilgili, A., Karaöz, E., Gökçimen, A., Eraslan, G. & Üstüner, E. 2002. The structural and biochemical changes of kidney tissue on fluorosis in rabbits. Turkish Journal of Veterinary and Animal Sciences, 26: 71-77.
• 2. Ameeramja, J., Panneerselvam, L., Govindarajan, V., Jeyachandran, S., Baskaralingam, V. & Perumal, E. 2016. Tamarind seed coat ameliorates fluoride induced cytotoxicity, oxidative stress, mitochondrial dysfunction and apoptosis in A549 cells. Journal of Hazardous Materials, 301: 554-565.
• 3. Ayoob, S. & Gupta, A.K. 2006. Fluoride in drinking water: a review on the status and stress effects. Critical Reviews in Environmental Science and Technology, 36: 433-487.
• 4. Baba, A. & Tayfur, G. 2011. Groundwater contamination and its effect on health in Turkey. Environmental Monitoring and Assessment, 183: 77-94.
• 5. Bataineh, H.N. & Nusier, M.K. 2006. Impact of 12-week ingestion of sodium fluoride on aggression, sexual behavior, and fertility in adult male rats. Fluoride, 39: 293-301.
• 6. Bhattacharya, S. 2015. Reactive oxygen species and cellular defense system. Pp. 17-29. In: Rani, V. & Singh Yadav, U.C. (eds) Free Radicals in Human Health and Disease. Springer, New Delhi, 427 pp.
• 7. Chachra, D., Limeback, H., Willett, T.L. & Grynpas, M.D. 2010. The long-term effects of water fluoridation on the human skeleton. Journal of Dental Research, 89: 1219-1223.
• 8. Chang, Y.-C. & Chou, M.-Y. 2001. Cytotoxicity of fluoride on human pulp cell cultures in vitro. Oral Surgery, Oral Medicine, Oral Pathology, Oral Radiology, and Endodontology, 91: 230-234.
• 9. Chinoy, N., Tewari, K. & Jhala, D. 2004. Fluoride and/or arsenic toxicity in mice testis with formation of giant cells and subsequent recovery by some antidotes. Fluoride, 37: 172-184.
• 10. Chouhan, S. & Flora, S.J. 2010. Arsenic and fluoride: two major ground water pollutants. Indian Journal of Experimental Biology, 48: 666-678.
• 11. Chouhan, S. & Flora, S.J.S. 2008. Effects of fluoride on the tissue oxidative stress and apoptosis in rats: Biochemical assays supported by IR spectroscopy data. Toxicology, 254: 61-67.
• 12. Chouhan, S., Lomash, V. & Flora, S.J. 2010. Fluoride-induced changes in haem biosynthesis pathway, neurological variables and tissue histopathology of rats. Journal of Applied Toxicology, 30: 63-73.
• 13. Devasagayam, T.P. & Tarachand, U. 1987. Decreased lipid peroxidation in the rat kidney during gestation. Biochemical and Biophysical Research Communications, 145: 134-138.
• 14. Dey, S. & Giri, B. 2016. Fluoride Fact on Human Health and Health Problems: A Review. Medical & Clinical Reviews 2(1-2): 1-6.
• 15. Dong, C., Cao, J., Cao, C., Han, Y., Wu, S., Wang, S. & Wang, J. 2016. Effects of fluoride and aluminum on expressions of StAR and P450scc of related steroidogenesis in guinea pigs' testis. Chemosphere, 147: 345-351.
• 16. Ellman, G.L. 1959. Tissue sulfhydryl groups. Archives of Biochemistry and Biophysics, 82: 70-77.
• 17. Fawell, J.K. Bailey, K., Chilton, J., Dahi, E., Fewtrell, L., & Magara, Y., 2006. Fluoride in drinking-water. World Health Organization.
• 18. Feng, D.M., Huang, H., Yang, Y., Yan, T., Jin, Y.F., Cheng, X.M. & Cui, L.X. 2015. Ameliorative effects of N-acetylcysteine on fluoride-induced oxidative stress and DNA damage in male rats' testis. Mutation Research-Genetic Toxicology and Environmental Mutagenesis, 792: 35-45.
• 19. Gupta, R., Khan, T., Agrawal, D. & Kachhawa, J. 2007. The toxic effects of sodium fluoride on the reproductive system of male rats. Toxicology and Industrial Health, 23: 507-513.
• 20. Habig, W.H., Pabst, M.J. & Jakoby, W.B. 1974. Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. Journal of Biological Chemistry, 249: 7130-7139.
• 21. Hafeman, D.G., Sunde, R.A. & Hoekstra, W.G. 1974. Effect of dietary selenium on erythrocyte and liver glutathione peroxidase in the rat. Journal of Nutrition, 104: 580-587.
• 22. Holland, M.K. & Storey, B.T. 1981. Oxygen metabolism of mammalian spermatozoa. Generation of hydrogen peroxide by rabbit epididymal spermatozoa. Biochemical Journal, 198: 273-280.
• 23. Izquierdo-Vega, J.A., Sanchez-Gutierrez, M. & Del Razo, L.M. 2008. Decreased in vitro fertility in male rats exposed to fluoride-induced oxidative stress damage and mitochondrial transmembrane potential loss. Toxicology and Applied Pharmacology, 230: 352-357.
• 24. Kaminsky, L.S., Mahoney, M.C., Leach, J., Melius, J. & Miller, M.J. 1990. Fluoride: benefits and risks of exposure. Critical Reviews in Oral Biology & Medicine, 1: 261-281.
• 25. Kim, J., Kwon, W.S., Rahman, M.S., Lee, J.S., Yoon, S.J., Park, Y.J., You, Y.A. & Pang, M.G. 2015. Effect of sodium fluoride on male mouse fertility. Andrology, 3: 544-551.
• 26. Kohen, R. & Nyska, A. 2002. Invited review: Oxidation of biological systems: oxidative stress phenomena, antioxidants, redox reactions, and methods for their quantification. Toxicologic Pathology, 30: 620-650.
• 27. Lennon, M., Whelton, H., O’Mullane, D. & Ekstrand, J. 2004. Rolling Revision of the WHO Guidelines for Drinking-Water Quality. World Health Organization.
• 28. Long, H., Jin, Y., Lin, M., Sun, Y., Zhang, L. & Clinch, C. 2009. Fluoride toxicity in the male reproductive system. Fluoride, 42: 260-276.
• 29. Lu, Z., Wang, S., Sun, Z., Niu, R. & Wang, J. 2014. In vivo influence of sodium fluoride on sperm chemotaxis in male mice. Archives of Toxicology, 88: 533-539.
• 30. Marklund, S. & Marklund, G. 1974. Involvement of the superoxide anion radical in the autoxidation of pyrogallol and a convenient assay for superoxide dismutase. European Journal of Biochemistry, 47: 469-474.
• 31. Mittal, M. & Flora, S.J. 2007. Vitamin E supplementation protects oxidative stress during arsenic and fluoride antagonism in male mice. Drug and Chemical Toxicology, 30: 263-281.
• 32. Mittal, M. & Flora, S.M. 2006. Effects of individual and combined exposure to sodium arsenite and sodium fluoride on tissue oxidative stress, arsenic and fluoride levels in male mice. Chemico-Biological Interactions, 162: 128-139.
• 33. Mosmann, T. 1983. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. Journal of Immunological Methods, 65: 55-63.
• 34. Orlowski, M. & Meister, A. 1965. Isolation of Gamma-Glutamyl Transpeptidase from Hog Kidney. Journal of Biological Chemistry, 240: 338-347.
• 35. Orta, B. & Erkan, M. 2014. Effects of Vitamin C on Antioxidant Systems and Steroidogenic Enzymes in Sodium Fluoride-Exposed Tm4 Sertoli Cells. Fluoride, 47: 139-151.
• 36. Oruc, N. 2008. Occurrence and problems of high fluoride waters in Turkey: an overview. Environmental Geochemistry and Health, 30: 315-323.
• 37. Puntarulo, S. & Cederbaum, A.I. 1988. Effect of oxygen concentration on microsomal oxidation of ethanol and generation of oxygen radicals. Biochemical Journal, 251: 787-794.
• 38. Rao, M.V. & Bhatt, R.N. 2012. Protective Effect of Melatonin on Fluoride-Induced Oxidative Stress and Testicular Dysfunction in Rats. Fluoride, 45: 116-124.
• 39. Riksen, E.A., Kalvik, A., Brookes, S., Hynne, A., Snead, M.L., Lyngstadaas, S.P. & Reseland, J.E. 2011. Fluoride reduces the expression of enamel proteins and cytokines in an ameloblast-derived cell line. Archives of Oral Biology, 56: 324-330.
• 40. Sinha, A. K. 1972. Colorimetric assay of catalase. Analitical Biochemistry, 47: 389-394.
• 41. Song, C., Cao, X., Yang, Z., Guo, S. & Shang, Z. 2013. Fluoride-induced cell cycle arrest, apoptosis, and DNA damage in TM3 mouse Leydig cells. Fluoride, 46: 218-223.
• 42. Song, G., Wang, R.L., Chen, Z.Y., Zhang, B., Wang, H.L., Liu, M.L., Gao, J.P. & Yan, X.Y. 2014. Toxic effects of sodium fluoride on cell proliferation and apoptosis of Leydig cells from young mice. Journal Physiology and Biochemistry, 70: 761-768.
• 43. Sun, W., Zhang, G., Tan, L., Yang, K. & Ai, H. 2016. The fluoride coated AZ31B magnesium alloy improves corrosion resistance and stimulates bone formation in rabbit model. Materials Science and Engineering C: Materials for Biological Applications, 63: 506-511.
• 44. Sun, Z., Niu, R., Su, K., Wang, B., Wang, J., Zhang, J. & Wang, J. 2010. Effects of sodium fluoride on hyperactivation and Ca2+ signaling pathway in sperm from mice: an in vivo study. Archives of Toxicology, 84: 353-361.
• 45. Wan, S.X., Zhang, J.H. & Wang, J.D. 2006. Effects of high fluoride on sperm quality and testicular histology in male rats. Fluoride, 39: 17-21.
• 46. Wang, A.G., Xia, T., Ru, R., Yuan, J., Chen, X.M., Yang, K.D. & Yang, K.D. 2004. Antagonistic effect of selenium on oxidative stress, DNA damage, and apoptosis induced by fluoride in human hepatocytes. Fluoride, 37: 107-116.
• 47. Wang, J., Zhang, Y., Zhang, H., Zhang, K., Zhang, Z. & Li, J. 2009. Toxic effects of fluoride on reproductive ability in male rats: sperm motility, oxidative stress, cell cycle, and testicular apoptosis. Fluoride, 42: 174.
• 48. Wang, L., Zhu, Y. & Wang, D. 2016. High-Dose Fluoride Induces Apoptosis and Inhibits Ameloblastin Secretion in Primary Rat Ameloblast. Biological Trace Element Research, 174: 402-409.
• 49. Weyermann, J., Lochmann, D. & Zimmer, A. 2005. A practical note on the use of cytotoxicity assays. International Journal of Pharmaceutics, 288: 369-376.
• 50. Yan, Q., Zhang, Y., Li, W. & Denbesten, P. 2007. Micromolar fluoride alters ameloblast lineage cells in vitro. Journal of Dental Research, 86: 336-340.
• 51. Yang, Y., Huang, H., Ba, Y., Cheng, X.M. & Cui, L.X. 2015. Effect of oxidative stress on fluoride-induced apoptosis in primary cultured Sertoli cells of rats. International Journal of Environmental Health Research, 25: 1-9.
• 52. Yang, Z.J., Song, C., Cao, X.D., Zhang, F.F., Zhao, H.X., & Du, B.Q. 2013. Fluoride-Induced Oxidative Damage and DNA-Protein Crosslinks in Tm3 Mouse Leydig Cells. Fluoride, 46: 212-217.
• 53. Yilmaz, B. O. & Erkan, M. 2015. Effects of Vitamin C on Sodium Fluoride-Induced Oxidative Damage in Sertoli Cells. Fluoride, 48: 241-251.
• 54. Ying, J., Xu, J., Shen, L., Mao, Z., Liang, J., Lin, S., Yu, X., Pan, R., Yan, C. & Li, S. 2017. The Effect of Sodium Fluoride on Cell Apoptosis and the Mechanism of Human Lung BEAS-2B Cells In Vitro. Biological Trace Element Research, doi:10.1007/s12011-017-0937-y, 1-11.
• 55. Zakrzewska, H., Udala, J. & Blaszczyk, B. 2002. In vitro influence of sodium fluoride on ram semen quality and enzyme activities. Fluoride, 35: 153-160.
• 56. Zhang, M., Wang, A., He, W., He, P., Xu, B., Xia, T., Chen, X. & Yang, K. 2007. Effects of fluoride on the expression of NCAM, oxidative stress, and apoptosis in primary cultured hippocampal neurons. Toxicology, 236: 208-216.
• 57. Zhang, S., Jiang, C.Y., Liu, H.L., Guan, Z.Z., Zeng, Q., Zhang, C., Lei, R.R., Xia, T., Gao, H., Yang, L., Chen, Y.H., Wu, X., Zhang, X.F., Cui, Y.S., Yu, L.Y., Wang, Z.L. & Wang, A.G. 2013. Fluoride-elicited developmental testicular toxicity in rats: Roles of endoplasmic reticulum stress and inflammatory response. Toxicology and Applied Pharmacology, 271: 206-215.