Shen Xing-Hui, Cui Xiang-Shun, Lee Sung-Hyun, Kim Nam-Hyung
Department of Histology and Embryology, Harbin Medical University, Heilongjiang Province, China.
J Reprod Dev. 2012;58(4):453-60. doi: 10.1262/jrd.2012-015. Epub 2012 Apr 21.
Signal transducer and activator of transcription-3 (Stat3) plays a central role in interleukin-6 (IL-6)-mediated cell proliferation by inhibiting apoptosis in a variety of cell types. The Stat3 pathway is essential for embryonic development. The aim of this study was to determine the effects of recombinant IL-6 on the viability and development of porcine diploid parthenotes cultured in vitro. Four-cell parthenotes, derived in vitro, were cultured to the blastocyst stage, with or without recombinant IL-6. The addition of 10 or 100 ng/ml of recombinant swine IL-6 into PZM3 medium increased the development rate of parthenotes to the blastocyst stage (P<0.05). When supplemented with 10 ng/ml of recombinant swine IL-6, the number of parthenotes at the blastocyst stage increased (P<0.05) and apoptosis decreased (P<0.05). Real-time RT-PCR experiments revealed that the addition of recombinant swine IL-6 decreased the mRNA expression of the pro-apoptotic gene Caspase3 (P<0.01) but increased the expression levels of the anti-apoptotic genes Bcl2l1 and Survivin. IL-6 receptors and Stat3 mRNA expression were upregulated after treatment with 10 ng/ml recombinant swine IL-6. Immunoblots and fluorescence labeling experiments showed that the levels of phosphorylated Stat3 were upregulated. These results suggest that recombinant swine IL-6 prevents apoptosis of porcine parthenotes and enhances porcine embryo viability through the IL-6/Stat3 signaling pathway in vitro.
信号转导与转录激活因子3(Stat3)通过抑制多种细胞类型的凋亡,在白细胞介素-6(IL-6)介导的细胞增殖中发挥核心作用。Stat3信号通路对胚胎发育至关重要。本研究的目的是确定重组IL-6对体外培养的猪二倍体孤雌胚胎的活力和发育的影响。将体外获得的四细胞孤雌胚胎在添加或不添加重组IL-6的情况下培养至囊胚阶段。向PZM3培养基中添加10或100 ng/ml的重组猪IL-6可提高孤雌胚胎发育至囊胚阶段的比率(P<0.05)。当添加10 ng/ml的重组猪IL-6时,囊胚阶段的孤雌胚胎数量增加(P<0.05),凋亡减少(P<0.05)。实时RT-PCR实验表明,添加重组猪IL-6可降低促凋亡基因Caspase3的mRNA表达(P<0.01),但增加抗凋亡基因Bcl2l1和Survivin的表达水平。用10 ng/ml重组猪IL-6处理后,IL-6受体和Stat3 mRNA表达上调。免疫印迹和荧光标记实验表明,磷酸化Stat3的水平上调。这些结果表明,重组猪IL-6在体外通过IL-6/Stat3信号通路防止猪孤雌胚胎凋亡并提高猪胚胎活力。