Luptakova Lenka, Dvorcakova Simona, Demcisakova Zuzana, Belbahri Lassaad, Holovska Katarina, Petrovova Eva
Department of Biology and Physiology, University of Veterinary Medicine and Pharmacy in Kosice, Komenskeho 73, 041 81 Kosice, Slovakia.
Department of Morphological Disciplines, University of Veterinary Medicine and Pharmacy in Kosice, Komenskeho 73, 041 81 Kosice, Slovakia.
Toxics. 2021 Mar 12;9(3):55. doi: 10.3390/toxics9030055.
Dimethyl sulfoxide (DMSO) is widely used as a solvent for small hydrophobic drug molecules. However, the safe volume allowing to avoid its embryotoxic effect has been poorly studied. In this study, we documented the effects of dimethyl sulfoxide (DMSO) in the developing chicken embryo at morphological, histological, and molecular levels. We focused on the developing chicken liver as the main organ involved in the process of detoxification. In our study, 100% DMSO was administered subgerminally onto the eggshell membrane (membrana papyracea) at various volumes (5, 10, 15, 20, 25, 30, 35, and 50 µL) on 4th embryonic day (ED). We focused on histopathological alterations of the liver structure, and noticed the overall impact of DMSO on developing chicken embryos (embryotoxicity, malformation). At the molecular level, we studied cytochrome P450 complex () isoform's activities in relation to changes of , , and gene expression. Total embryotoxicity after application of different doses of DMSO on ED 4, and the embryo lethality increased with increasing DMSO amounts. Overall mortality after DMSO administration ranged below 33%. Mortality was increased with higher amounts of DMSO, mainly from 20 µL. The highest mortality was observed for the highest dose of DMSO over 35 µL. The results also showed a decrease in body weight with increased application volumes of DMSO. At the histological level, we observed mainly the presence of lipid droplets and dilated bile canaliculi and sinusoids in samples over the administration of 25 µL of DMSO. While these findings were not statistically significant, DMSO treatment caused a significant different up-regulation of mRNA expression in all studied genes. For , , and DMSO volumes needed were 15 µL, 10 µL, and 20 µL, respectively. A significant down-regulation of all studied isoform was detected after application of a DMSO dose of 5 µL. Regarding the morphological results, we can assume that the highest safe dose of DMSO without affecting chicken embryo development and its liver is up to 10 µL. This conclusion is corroborated with the presence of number of malformations and body weight reduction, which correlates with histological findings. Moreover, the gene expression results showed that even the lowest administered DMSO volume could affect hepatocytes at the molecular level causing down-regulation of cytochrome P450 complex (, , ).
二甲基亚砜（DMSO）被广泛用作小分子疏水性药物的溶剂。然而，关于避免其胚胎毒性作用的安全用量研究较少。在本研究中，我们从形态学、组织学和分子水平记录了二甲基亚砜（DMSO）对发育中的鸡胚的影响。我们将发育中的鸡肝脏作为参与解毒过程的主要器官进行研究。在我们的研究中，于胚胎第4天（ED）将100%的DMSO以不同体积（5、10、15、20、25、30、35和50微升）经胚下注射到卵壳膜（卵壳纸膜）上。我们重点关注肝脏结构的组织病理学改变，并注意到DMSO对发育中的鸡胚的总体影响（胚胎毒性、畸形）。在分子水平上，我们研究了细胞色素P450复合物（）同工型的活性与、和基因表达变化的关系。在胚胎第4天应用不同剂量的DMSO后，总体胚胎毒性以及胚胎致死率随DMSO用量的增加而升高。DMSO给药后的总体死亡率低于33%。较高剂量的DMSO会增加死亡率，主要是从20微升开始。观察到超过35微升的最高剂量DMSO导致的死亡率最高。结果还表明，随着DMSO应用体积的增加，鸡胚体重下降。在组织学水平上，我们观察到在给予25微升DMSO以上的样本中主要存在脂滴以及扩张的胆小管和肝血窦。虽然这些发现无统计学意义，但DMSO处理导致所有研究基因的mRNA表达出现显著不同程度的上调。对于、和，所需的DMSO体积分别为15微升、10微升和20微升。在应用5微升DMSO剂量后，检测到所有研究的同工型均有显著下调。关于形态学结果，我们可以假设不影响鸡胚发育及其肝脏的DMSO最高安全剂量为10微升。这一结论得到了畸形数量和体重减轻的支持，这与组织学结果相关。此外，基因表达结果表明，即使是最低剂量的DMSO也可能在分子水平上影响肝细胞，导致细胞色素P450复合物（、、）下调。
