Department of Marine Biology, Faculty of Marine Sciences, Khorramshahr University of Marine Science and Technology, Khorramshahr, Iran.
Department of Biology, Payame Noor University, Tehran, Iran.
J Appl Toxicol. 2020 Jul;40(7):991-1003. doi: 10.1002/jat.3958. Epub 2020 Feb 26.
We assessed the toxic effects of benzo[a]pyrene (BaP) on cell viability, aromatase (Aro) activity and steroid production using ovarian and brain cell cultures obtained from Mullet, Liza klunzingeri. The brain and ovary were minced and digested, and the cells were suspended in Leibovitz's L-15 medium supplemented with 15% and 20% fetal bovine serum. The cell suspensions were seeded on 25-cm cell-culture flasks at 1 × 10 cells/mL and incubated at 25 °C for 2 weeks. A BaP concentration of 10 mol/L was accepted as the half-maximal inhibitory concentration. Ovarian and brain cells were exposed to different concentrations of BaP [0 (control), 10 , 2 × 10 , 3 × 10 mol/L] and incubated at 30 °C. At different sampling times (0, 12, 24 and 48 h) 40 ng/10 cells of 1,4,6-androstatriene-3,17-dione (ATD) was added to each well. Aro activity, 17β-estradiol (E2) and ATD production were determined. The sensitivity of the cultivated ovarian and brain cells to BaP increased dose dependently. BaP was a potent inhibitor of Aro activity at 2 × 10 and 3 × 10 mol/L, both in the cultivated brain and ovarian cells at different sampling times, with 10 mol/L BaP found to be the least potent Aro inhibitor. E2 production decreased from cultivated ovarian and brain cells treated by different concentrations of BaP. In conclusion, BaP is able to change the activity of Aro and disrupt the biosynthesis of estrogens, and thus affects reproduction in fish.
我们评估了苯并[a]芘(BaP)对细胞活力、芳香酶(Aro)活性和类固醇产生的毒性作用,使用从 Mullet,Liza klunzingeri 获得的卵巢和脑细胞培养物。将脑和卵巢切成小块并消化,将细胞悬浮在补充有 15%和 20%胎牛血清的 Leibovitz 的 L-15 培养基中。将细胞混悬液以 1×10 个细胞/mL 的浓度接种到 25-cm 细胞培养瓶中,并在 25°C 孵育 2 周。10 mol/L 的 BaP 浓度被接受为半最大抑制浓度。将卵巢和脑细胞暴露于不同浓度的 BaP[0(对照)、10、2×10、3×10 mol/L]并在 30°C 下孵育。在不同的采样时间(0、12、24 和 48 h)时,向每个孔中添加 40 ng/10 个细胞的 1,4,6-雄三烯-3,17-二酮(ATD)。测定 Aro 活性、17β-雌二醇(E2)和 ATD 产量。培养的卵巢和脑细胞对 BaP 的敏感性呈剂量依赖性增加。BaP 是一种有效的 Aro 活性抑制剂,在 2×10 和 3×10 mol/L 时,在不同采样时间的培养脑和卵巢细胞中均如此,而 10 mol/L BaP 被发现是最不有效的 Aro 抑制剂。不同浓度的 BaP 处理后,培养的卵巢和脑细胞的 E2 产量减少。总之,BaP 能够改变 Aro 的活性并破坏雌激素的生物合成,从而影响鱼类的繁殖。