Department of Animal Genetics, Breeding and Reproduction, College of Animal Science and Technology, Northeast Agricultural University, Harbin, People's Republic of China.
Department of Animal Science and Technology and BET Research Institute, Chung-Ang University, Anseong, South Korea.
Biol Trace Elem Res. 2022 Mar;200(3):1140-1155. doi: 10.1007/s12011-021-02731-0. Epub 2021 Apr 24.
This study investigated the antioxidant role of selenium (Se) in the form of selenomethionine (SLM) in LPS-induced oxidative stress via the glutathione peroxidase (GPx) enzymes and the Nrf2/HO-1 transcription factor. The impact of serum supplementation in culture media on GPxs was also studied. The bovine uterus is constantly exposed to exogenous pathogens postpartum, and the endometrium is the first contact against bacteria invasion. Endometritis is an inflammation of the endometrium and is brought about by bacterial lipopolysaccharide capable of inducing oxidative stress. The BEND cells were supplemented at the point of seeding with the following SLM concentrations 0, 100, 500, and 1000 nM for 48 h. BEND cells, cultured with or without SLM (100 nM), were initially incubated for 48 h, and then, we serum starved the SLM group for 24, 48, and 72 h. Similarly, an assay involving serum volume (0, 2, 5, and 10%) supplementation in culture media (v/v) with or without SLM (100 nM) was performed for 48 h. The BEND cells were also seeded into four experimental groups and cultured for an initial 48 h as follows: control, LPS (20 μg/mL), SLM (100 nM), and SLM + LPS groups followed by 6-h LPS treatment. The role of SLM in modulating the expressions of GPx1 and GPx4 and the Nrf2 transcription factor-related genes was assessed using qRT-PCR and Western blot techniques. The results showed serum starvation in the presence of SLM supplementation decreased the expression of GPx1 enzyme but increased GPx4 compared to the control. The addition of SLM to cell culture media in an FBS limiting condition improved the expressions of both GPx1 and GPx4. SLM supplementation promoted GPx enzymes' expressions in a serum-free media (0%) and at 2% FBS in media. However, it did not improve their expressions at 10% FBS in media than the untreated groups. Together, our data show the protective role of Se by regulating the expressions of GPx1 and GPx4 enzymes in BEND cells. It also shows that SLM promoted the expression of Nrf2 transcription factor-related genes at both the mRNA and protein levels in BEND cells during LPS stimulation.
本研究通过谷胱甘肽过氧化物酶 (GPx) 酶和 Nrf2/HO-1 转录因子研究了硒 (Se) 以硒蛋氨酸 (SLM) 的形式在 LPS 诱导的氧化应激中的抗氧化作用。还研究了血清在培养基中的补充对 GPxs 的影响。牛子宫在产后不断暴露于外源性病原体，子宫内膜是抵抗细菌入侵的第一道防线。子宫内膜炎是子宫内膜的炎症，由能够诱导氧化应激的细菌脂多糖引起。BEND 细胞在接种时补充以下浓度的 SLM0、100、500 和 1000 nM，持续 48 小时。用或不用 SLM（100 nM）培养 BEND 细胞 48 小时，然后将 SLM 组血清饥饿 24、48 和 72 小时。同样，还进行了一项涉及在含有或不含 SLM（100 nM）的培养基中补充血清量（0、2、5 和 10%）（v/v）的测定，持续 48 小时。BEND 细胞还接种到四个实验组中，初始培养 48 小时，如下所示：对照组、LPS（20 μg/mL）、SLM（100 nM）和 SLM+LPS 组，然后用 LPS 处理 6 小时。使用 qRT-PCR 和 Western blot 技术评估 SLM 在调节 GPx1 和 GPx4 酶和 Nrf2 转录因子相关基因表达中的作用。结果表明，在存在 SLM 补充的情况下血清饥饿降低了 GPx1 酶的表达，但与对照组相比，GPx4 的表达增加。在 FBS 限制条件下将 SLM 添加到细胞培养基中可提高 GPx1 和 GPx4 的表达。SLM 补充在无血清培养基（0%）和培养基中 2% FBS 中促进 GPx 酶的表达。然而，它在培养基中 10% FBS 中改善其表达的作用不如未处理组。总之，我们的数据表明，Se 通过调节 BEND 细胞中 GPx1 和 GPx4 酶的表达发挥保护作用。它还表明，在 LPS 刺激期间，SLM 在 mRNA 和蛋白质水平上均促进了 BEND 细胞中 Nrf2 转录因子相关基因的表达。
