Division of Animal Biotechnology, College of Animal Life Science, Kangwon National University, Chuncheon, Republic of Korea.
Cryobiology. 2010 Apr;60(2):211-6. doi: 10.1016/j.cryobiol.2009.12.002. Epub 2009 Dec 16.
Plasminogen activators (PAs) play a pivotal role in a variety of uterine physiologies, such as endometrial function, trophoblast invasion, and implantation process, but its alteration in expression or activity during cryopreservation of primary uterine cells has received little attention. In this study, we investigated whether PA expression and activity were modulated in first passage primary porcine uterus endometrial epithelium cells (PUEECs) treated with or without a freezing-thawing procedure. Western blotting and zymographic analysis showed that uPA expression and activity increased significantly in frozen-thawed PUEECs in a passage-dependent manner as compared to freshly prepared control cells. Moreover, intracellular reactive oxygen species (ROS) were increased by freezing-thawing and longer culturing, and were more prominent in frozen-thawed PUEECs than in control cells. However, the increase in both uPA expression and activity was greatly reduced or alleviated by treatment with either ROS scavenger N-acetylcysteine or extracellular signal-regulated kinase (ERK) inhibitor PD98059. These results suggest that ROS/ERK-mediated uPA activation may be an important factor in cryo-damage of primary uterine cells.
纤溶酶原激活物（PAs）在多种子宫生理学中发挥着关键作用，如子宫内膜功能、滋养细胞浸润和着床过程，但在原代子宫细胞的冷冻保存过程中，其表达或活性的改变尚未得到充分关注。在这项研究中，我们研究了是否在冷冻-解冻处理的情况下，PA 的表达和活性在第一传代的猪子宫子宫内膜上皮细胞（PUEECs）中被调节。Western blot 和酶谱分析显示，与新鲜制备的对照细胞相比，冷冻-解冻后的 PUEECs 中 uPA 的表达和活性显著增加，且具有传代依赖性。此外，冷冻-解冻和延长培养会增加细胞内的活性氧（ROS），且在冷冻-解冻后的 PUEECs 中比对照细胞更为明显。然而，用 ROS 清除剂 N-乙酰半胱氨酸或细胞外信号调节激酶（ERK）抑制剂 PD98059 处理后，uPA 的表达和活性的增加明显减少或缓解。这些结果表明，ROS/ERK 介导的 uPA 激活可能是原代子宫细胞冷冻损伤的一个重要因素。
