Dibutyl Phthalate (DBP) Mediated Oxidative Stress, Cytotoxicity and Estradiol Synthesis in Cultured Ovine Granulosa Cell
Keywords:
Biochemical assay, Dibutyl phthalate, Estradiol, Oxidative stress, SheepAbstract
Plasticizer Dibutyl phthalate (DBP), a potent reproductive toxicant, can cause damage to the granulosa cells, the key cells for the production of steroidogenic hormones. It may also affect other antioxidant enzymes for follicular development, affecting oocyte quality. The present study examined the DBP-induced oxidative stress and hormone synthesis from granulosa cells. Biochemical assays such as MTT (3- (-4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide ) for the study of cell viability, proliferation, and cytotoxicity, cupric reducing antioxidant capacity (CUPRAC) assay for the total antioxidant capacity, malondialdehyde (MDA) assay for the lipid peroxidation, and Enzyme-linked immunosorbent assay (ELISA) for estradiol synthesis were employed to evaluate the effect of DBP at a concentration of 0 (control), 1, 10, 25, 50, 100 µM for 5 days in cultured bovine granulosa cells. Our results revealed that DBP decreased cell viability, cell proliferation, induced oxidative cellular stress, and affected estradiol secretion in a non-dose-dependent manner. The lowest concentration of DBP (1 µM) had a significant decrease in cell viability and increased lipid peroxidation compared to those observed in the control groups. The total antioxidant enzyme (CUPRAC) activity was non significantly induced in all concentrations of DBP in treated granulosa cells. The estradiol synthesis was increased significantly as the DBP concentrations increased. This study suggested that DBP at a very low concentration affected the steroidogenic capacity, cell viability, and proliferation by altering the antioxidant enzyme activity and causing oxidative stress to the cell.
Downloads
References
Ahmed, T., Pathak, R., Mustafa, M.D., Kar, R., Tripathi, A.K., Ahmed, R.S., & Banerjee, B.D. (2011). Ameliorating effect of N-acetylcysteine and curcumin on pesticide-induced oxidative DNA damage in human peripheral blood mononuclear cells. - Environmental Monitoring and Assessment. 179,293–309.
ATSDR. (2001). Toxicological Profile for di-n-butyl phthalate (DBP). Atlanta, GA. Available online at: http://www.atsdr.cdc.gov/ toxprofiles.
Catherine, A., Pirkko, H., Malcolm, G. P., & Sumpter, J. P. (1997). The Estrogenic Activity of Phthalate Esters in Vitro. Annals of Clinical & Laboratory Science,105,802-811.
El-Deeb , I.M., Yoo, K., & Lee, H. (2010). ROS receptor tyrosine kinase: a new potential target for anticancer drugs. Medical Care Research and Review. 31,794–818.
Erkekoglu, P., Rachidi, W., Yuzugullu, O.G., Giray, B., Ozturk, M., Favier, A., & Hincal, F. (2011). Induction of ROS, p53, p21 in DEHP- and MEHP-exposed LNCaP cells-protection by selenium compounds. Food and Chemical Toxicology. 49,1565–1571. Fennell, T., Krol, W., Sumner, S., & Snyder, R. (2004).Pharmacokinetics of dibutyl phthalate in pregnant rats. Toxicological Sciences. 82, 407–418.
Guo, Z.Y., Gai, P.P, Duan, J., Zhai, J.X., Zhao, S.S., Wang, S., & Wei, D.Y. (2010). Simultaneous determination of phthalates and adipates in human serum using gas chromatography-mass spectrometry with solid-phase extraction. Biomedical Chromatography. 24,1094-1099.
Hu, J., Du, G., Zhang, W., Huang, H., Chen, D., Wu, D., Wang, X. (2013). Short-term neonatal/prepubertal exposure of dibutyl phthalate (DBP) advanced pubertal timing and affected hypothalamic kisspeptin/GPR54 expression differently in female rats. Toxicology. 314, 65-75.
Jakub, K., Yoshihiro, N., Toshihiko, Y., & Giudice, L. (2010). Peroxisome Proliferator-Activated Receptor-Mediates Bisphenol A Inhibition of FSH-Stimulated IGF-1, Aromatase, and Estradiol in Human Granulosa Cells. Environmental Health Perspectives.118, 400-406.
Knock, GA, & Ward, J.P. (2011). Redox regulation of protein kinases as a modulator of vascular function. Antioxidant Redox Signal. 15,1531-1547.
Latini, G. (2005). Monitoring phthalate exposure in humans. Clinica Chimica Acta. 361, 20–29.
Lin, K.H.R., Tsou, C.C., Hwang, S.Y., Chen, L.F.O., & Lo, H.F. (2006). Paclobutrazol pretreatment enhanced flooding tolerance of sweet potato. Journal of Plant Physiology. 163,750– 760.
Lovekamp-Swan, T., & Davis, B. J. (2003). Mechanisms of phthalate ester toxicity in the female reproductive system. Environmental Health Perspectives. 111, 139–145.
Nandi, S., Gupta, P.S.P., & Mondal, S. (2016). Ammonia concentrations in different size classes of ovarian follicles of sheep (Ovis aries): Possible mechanisms of accumulation and its effect on oocyte and granulosa cell growth in vitro. Theriogenology. 85, 678–
Parveen, M., Inoue, A., Ise, , R., Tanji, M. & Kiyama, R. (2008). Evaluation of estrogenic activity of phthalate esters by gene expression profiling using a focused microarray. Environmental Toxicology & Chemistry. 27, 1416-1425.
Rhind S.M. (2002). Endocrine-disrupting compounds and farm animals: their properties, actions, and routes of exposure. Domestic Animal Endocrinology. 23, 179–187.
Roszak, J., Smokpieniazek, A., Domeradzkagajda, K., Grobelny, J., Tomaszewska , E., Ranosze-ksoliwoda., Celichowski, K., & Stepnik, M. G. (2017). Inhibitory effect of silver nanoparticles on proliferation of estrogen-dependent MCF-7/BUS human breast cancer cells induced by butylparaben or di-n-butyl phthalate. Toxicology and Applied Pharmacology. 337.
Silva, M. J., Reidy, J. A., Herbert, A. R., Preau, J. L., Needham, L. L., & Calafat, A.M. (2004). Detection of phthalate metabolites in human amniotic fluid. Bulletin of Environmental Contamination and Toxicology. 72, 1226–1231.
Vandenberg, L., Colborn, N.T., Hayes, T.B., Heindel, J.J., Lee, J.D., Shioda, T. A., Soto, M., Vomsaal, F.S. & Welshons, W.V. (2012): Hormones and endocrine-disrupting chemicals: low-dose effects and non-monotonic dose responses. Endocrine Reviews.33, 378-455.
Welshons, W., Thayer, V.K.A., Judy, B. M., Taylor, J.A., Curran, E.M. & VomSaal, F.S. (2003). Large effects from small exposures. Mechanisms for endocrine-disrupting chemicals with estrogenic activity. Environmental Health Perspectives. 111, 994-1006.
Yaxian, M., Jun, Z., Rujun, Z., Xiaoyong, Q., Ran, C., Ying, N., Yunyao, L., Sicong, Li., Jing, Z.,Wenming, X., Liangzhi, X. & Ying, H. (2019). Effects of the Dibutyl Phthalate (DBP) on the Expression and Activity of Aromatase in Human Granulosa Cell Line KGN. Annals of Clinical & Laboratory Science, 49.
Downloads
Published
Issue
Section
License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
