Department of Medical Research, Division of High Risk Pregnancy, and Division of Infertility and Human Reproduction, Mackay Memorial Hospital, Taipei, Taiwan.
Biol Reprod. 2010 May;82(5):905-13. doi: 10.1095/biolreprod.109.081828. Epub 2010 Jan 27.
Reactive oxygen species may cause oxidative damage in the placenta, yet some mechanisms must exist to reduce or prevent such damage. We investigated whether oxidative injury to placental endothelial cells is inhibited by activation of antioxidant enzymes by paracrine factors secreted by human placental multipotent mesenchymal stromal cells (hPMSC). hPMSC-conditioned medium and umbilical endothelial cells were assayed for cytokines and cytokine receptor expression by immunoassay and real-time PCR. Endothelial cell survival was evaluated by MTS [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt] assay and caspase 3 activity assay. tert-Butyl hydroperoxide was used to induce oxidative injury in endothelial cells, with fluorescent microscopy and flow cytometry used to detect intracellular peroxides and cell apoptosis. Western blot, real-time PCR, STAT3 DNA-binding activity assay, and STAT3 siRNA were used to assess endothelial cell antioxidant enzymes. hPMSC-conditioned medium supported endothelial cell survival and reduced endothelial cell intracellular peroxides and apoptosis. hPMSCs expressed the transcripts of the interleukin (IL) 6 cytokine family, including IL6 and leukemia-inhibitory factor. hPMSC-conditioned medium activated STAT3 expression in endothelial cells, which was inhibited by neutralizing antibody to interleukin 6 signal transducer (IL6ST) but not to IL6 or leukemia-inhibitory factor. STAT3 siRNA or manganese superoxide dismutase (SOD2) siRNA transfected into endothelial cells inhibited the antiapoptotic effect of conditioned medium. SOD2 was significantly upregulated in endothelial cells by conditioned medium via STAT3 activation that, in turn, was inhibited by IL6ST-neutralizing antibody or STAT3 siRNA. Paracrine factors secreted by hPMSCs support endothelial cell survival. STAT3 activation and SOD2 production protect against oxidative stress-induced endothelial cell damage.
活性氧可能会导致胎盘氧化损伤，但肯定存在一些机制可以减少或防止这种损伤。我们研究了人胎盘多能间充质基质细胞（hPMSC）旁分泌因子激活抗氧化酶是否可以抑制胎盘内皮细胞的氧化损伤。通过免疫测定和实时 PCR 检测 hPMSC 条件培养基和脐内皮细胞中细胞因子和细胞因子受体的表达。通过 MTS [3-(4,5-二甲基噻唑-2-基）-5-(3-羧基甲氧基苯基）-2-(4-磺基苯基）-2H-四唑，内盐] 测定法和 caspase 3 活性测定法评估内皮细胞的存活率。用过氧化氢诱导内皮细胞氧化损伤，用荧光显微镜和流式细胞术检测细胞内过氧化物和细胞凋亡。使用 Western blot、实时 PCR、STAT3 DNA 结合活性测定和 STAT3 siRNA 评估内皮细胞抗氧化酶。hPMSC 条件培养基支持内皮细胞存活并减少内皮细胞内过氧化物和细胞凋亡。hPMSCs 表达白细胞介素（IL）6 细胞因子家族的转录本，包括 IL6 和白血病抑制因子。hPMSC 条件培养基激活内皮细胞中的 STAT3 表达，该表达可被白细胞介素 6 信号转导物（IL6ST）中和抗体抑制，但不能被 IL6 或白血病抑制因子抑制。STAT3 siRNA 或锰超氧化物歧化酶（SOD2）siRNA 转染入内皮细胞可抑制条件培养基的抗凋亡作用。通过条件培养基激活 STAT3 显著上调内皮细胞中的 SOD2，而 IL6ST 中和抗体或 STAT3 siRNA 则抑制该作用。hPMSC 旁分泌因子支持内皮细胞存活。STAT3 激活和 SOD2 产生可保护内皮细胞免受氧化应激诱导的损伤。
