Department of Reproductive Medicine, Shanghai First Maternity and Infant Hospital, Tongji University School of Medicine, Shanghai, 201204, China.
Department of Reproductive Medicine, Shanghai First Maternity and Infant Hospital, Tongji University School of Medicine, Shanghai, 201204, China.
Biochem Biophys Res Commun. 2019 Oct 20;518(3):423-429. doi: 10.1016/j.bbrc.2019.07.118. Epub 2019 Aug 21.
Oxidative stress is associated with functional disorder of trophoblast cells. Our previous studies have demonstrated that cyclosporin A (CsA) promotes the activity of normal human trophoblast cells. We further investigated the role and mechanism of CsA on oxidative stress in trophoblast cells. JEG-3 cells were co-cultured with HO and CsA. Cell viability and morphology were measured by MTT assay and inverted microscope. Reactive oxygen species (ROS) was analyzed by fluorescence microscopy. Cell mitochondrial membrane potential (MMP) was determined by flow cytometric analysis. Malondialdehyde (MDA) production, superoxide dismutase (SOD) and catalase (CAT) activities were examined using colorimetric assays. The expression and phosphorylation of FAK and Src kinase proteins were examined by western blotting. CsA increased JEG-3 cell viability and reduced the morphologic injury induced by HO treatment. CsA decreased ROS and MDA production, increased SOD and CAT activities, and restored the MMP of HO treated JEG-3 cells. CsA administration suppressed HO-induced reduction of FAK and Src phosphorylation. Blocking the activation of FAK or Src attenuated the protective effect of CsA on JEG-3 cells in HO-induced oxidative injury. CsA protects JEG-3 cells from HO-induced oxidative injury, and the FAK/Src signaling pathway plays an important role in this process.
氧化应激与滋养层细胞功能障碍有关。我们之前的研究表明环孢素 A（CsA）能促进正常人类滋养层细胞的活性。我们进一步研究了 CsA 在滋养层细胞氧化应激中的作用和机制。用 HO 和 CsA 共同培养 JEG-3 细胞。通过 MTT 检测和倒置显微镜观察细胞活力和形态。用荧光显微镜分析活性氧（ROS）。用流式细胞术分析细胞线粒体膜电位（MMP）。用比色法检测丙二醛（MDA）生成、超氧化物歧化酶（SOD）和过氧化氢酶（CAT）活性。用 Western blot 检测 FAK 和 Src 激酶蛋白的表达和磷酸化。CsA 增加 JEG-3 细胞活力，减轻 HO 处理引起的形态损伤。CsA 降低 ROS 和 MDA 生成，增加 SOD 和 CAT 活性，恢复 HO 处理的 JEG-3 细胞的 MMP。CsA 抑制 HO 诱导的 FAK 和 Src 磷酸化减少。阻断 FAK 或 Src 的激活可减弱 CsA 在 HO 诱导的氧化损伤中对 JEG-3 细胞的保护作用。CsA 可保护 JEG-3 细胞免受 HO 诱导的氧化损伤，FAK/Src 信号通路在此过程中起重要作用。
