Department of Animal Sciences, College of Agriculture, Isfahan University of Technology, Isfahan, 84156-83111, Iran.
Department of Animal Sciences, College of Agriculture, Isfahan University of Technology, Isfahan, 84156-83111, Iran.
Theriogenology. 2020 Feb;143:139-147. doi: 10.1016/j.theriogenology.2019.12.008. Epub 2019 Dec 16.
The present study was conducted to evaluate the effects of alpha-lipoic acid (ALA) on quantitative and qualitative indices of mouse embryos challenged by lipopolysaccharide (LPS). Having determined the effective concentrations of LPS (1 mg/mL) that could reduce blastocyst formation rate by around 50% and the optimal concentration of ALA (10 μM) that could attenuate the toxic effects of LPS on blastocyst formation, the following indices were defined: inner cell mass and trophectoderm cell numbers, blastocyst mitochondrial distribution, ROS and GSH levels, as well as the relative expression of Tlr-4. Nrf-2 and Tnf-RI/P-60 receptor involved in inflammatory pathways. Finally, embryos derived from the experimental and control groups were transferred to synchronized recipients and their implantation rate and post-implantation capacity were determined. Treatment with LPS resulted in an increase in intracellular ROS level (P ≤ 0.05), and remarkable decreases (P ≤ 0.05) in intracellular GSH content, mitochondrial mass, and blastocyst quality. ALA attenuated all the aforementioned negative effects of LPS. The relative expression levels of Nrf-2 and Tnf-RI/P-60 receptor (P ≤ 0.05) significantly increased in response to LPS, and treatment with ALA significantly reduced the relative expression of Tnf-RI/P-60. ALA also improved the post-implantation developmental capacity of embryos treated with LPS. In conclusion, our findings indicate that the reproductive toxicity of LPS could be overcome by ALA treatment. These effects were mainly due to the improvements made in intracellular antioxidant capacity as well as suppression of some inflammatory elements, especially the main receptor of TNF-α, the Tnf-RI/P-60, involved in induction of apoptosis. These observations have important implications for dairy farming and treatment of infertility.
本研究旨在评估α-硫辛酸（ALA）对脂多糖（LPS）刺激的小鼠胚胎数量和质量指标的影响。确定了有效浓度的 LPS（1mg/mL），它可以降低囊胚形成率约 50%，以及最佳浓度的 ALA（10μM），它可以减轻 LPS 对囊胚形成的毒性作用。定义了以下指标：内细胞团和滋养外胚层细胞数量、囊胚线粒体分布、ROS 和 GSH 水平，以及参与炎症途径的 TLR-4、Nrf-2 和 TNF-RI/P-60 受体的相对表达。最后，将来自实验组和对照组的胚胎移植到同步受体中，并确定其植入率和植入后能力。LPS 处理导致细胞内 ROS 水平升高（P≤0.05），细胞内 GSH 含量、线粒体质量和囊胚质量显著降低（P≤0.05）。ALA 减轻了 LPS 的所有上述负面影响。Nrf-2 和 TNF-RI/P-60 受体的相对表达水平（P≤0.05）因 LPS 而显著增加，而 ALA 处理显著降低了 TNF-RI/P-60 的相对表达。ALA 还改善了 LPS 处理的胚胎的植入后发育能力。总之，我们的研究结果表明，LPS 的生殖毒性可以通过 ALA 治疗来克服。这些作用主要归因于细胞内抗氧化能力的提高以及抑制某些炎症因子，特别是 TNF-α的主要受体 TNF-RI/P-60，其参与诱导凋亡。这些观察结果对奶牛养殖和不孕不育治疗具有重要意义。
