Laboratory of Histology and Embryology, Research Unit N°17/ES/13, Faculty of Medicine of Tunis, University of Tunis El Manar (UTM), Jabbari Jebel Lakhdar Street 15, 1007, Tunis, Tunisia.
Department of Microscopy, Laboratory of Cell Biology, Unit for Multidisciplinary Research in Biomedicine (UMIB), Institute of Biomedical Sciences Abel Salazar (ICBAS), University of Porto, R. de Jorge Viterbo Ferreira 228, 4050-013, Porto, Portugal; Universidade da Beira Interior, R. Marquês d'Ávila e Bolama, 6201-001, Covilhã, Portugal; LAQV/REQUIMTE - Laboratory of Bromatology and Hydrology, Faculty of Pharmacy, University of Porto, 4050-313, Porto, Portugal.
Theriogenology. 2019 Dec;140:188-200. doi: 10.1016/j.theriogenology.2019.08.030. Epub 2019 Aug 26.
Despite conflicting data on doxorubicin (DOX) reproductive toxicity, its chemotherapeutic potential sustains its use to treat different types of cancer. This work was designed to study the protective effect of a newly synthesized thiocyanoacetamide (TA), in comparison with selenium (Se), against doxorubicin-induced in vitro toxicity in rat Sertoli cells (SCs). DOX was administered alone or in combination with Se or TA. The possible protective role of increased concentrations of TA (0.25, 0.5 and 1 mM) or Se (12, 25 and 50 μM) on SCs was tested against 1 μM of DOX. From this screening, only the least toxic doses of TA and Se were used for further analysis. DOX cytotoxicity, as well as its impact on SCs viability, mitochondrial membrane potential (ΔΨ), oxidative stress biomarkers, apoptosis and autophagy were assessed. Our results showed that DOX exerted its cytotoxic effect through a significant increase in cell death. DOX-mediated cell death was not related to autophagy nor to an overproduction of reactive oxygen species. It was rather due to apoptosis, as shown by the increased number of apoptotic cells and increased activity of caspase-3, or due to necrosis, as shown by the increase in lactate dehydrogenase (LDH) extracellular activity. Still, Bax and Bcl-2 protein expression levels, as well as ΔΨ were not altered by the different treatments. Some individual doses of Se or TA induced a significant toxicity in SCs, however, when combined with DOX, there was a decrease in cell death, LDH extracellular activity, number of apoptotic cells and caspase-3 activity. Overall, our results indicate that DOX-mediated apoptosis in cultured SCs can possibly be averted through its association with specific doses of Se or TA. Nevertheless, TA showed a higher efficiency than Se in reducing DOX-induced toxicity in SCs by decreasing not only apoptosis, but also necrosis and autophagy.
尽管蒽环类药物（DOX）的生殖毒性存在矛盾的数据，但它的化疗潜力使其仍被用于治疗不同类型的癌症。本工作旨在研究一种新合成的硫氰酸乙酰胺（TA）与硒（Se）相比，对大鼠支持细胞（SCs）中 DOX 诱导的体外毒性的保护作用。DOX 单独或与 Se 或 TA 联合给药。TA（0.25、0.5 和 1mM）或 Se（12、25 和 50μM）的浓度增加可能对 SCs 具有保护作用，在 1μM DOX 下进行了测试。从该筛选中，仅使用毒性最低的 TA 和 Se 剂量进行进一步分析。评估了 DOX 的细胞毒性以及对 SCs 活力、线粒体膜电位（ΔΨ）、氧化应激生物标志物、细胞凋亡和自噬的影响。我们的结果表明，DOX 通过显著增加细胞死亡来发挥其细胞毒性作用。DOX 介导的细胞死亡与自噬或活性氧的过度产生无关，而是由于细胞凋亡，如凋亡细胞数量增加和 caspase-3 活性增加，或由于坏死，如乳酸脱氢酶（LDH）细胞外活性增加。然而，不同处理并未改变 Bax 和 Bcl-2 蛋白表达水平以及 ΔΨ。Se 或 TA 的一些单独剂量会导致 SCs 产生显著毒性，但与 DOX 联合使用时，细胞死亡、LDH 细胞外活性、凋亡细胞数量和 caspase-3 活性均降低。总的来说，我们的结果表明，DOX 介导的培养 SC 中的细胞凋亡可能通过其与特定剂量的 Se 或 TA 联合来避免。然而，与 Se 相比，TA 降低 DOX 诱导的 SC 毒性的效率更高，不仅降低了细胞凋亡，还降低了细胞坏死和自噬。
