Proteomic Analysis of Proteins Differentially Expressed in Uterine Lymphocytes Obtained From Wild-Type and NOD Mice
Proteomic Analysis of Proteins Differentially Expressed in Uterine Lymphocytes Obtained From Wild-Type and NOD Mice
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野生型和 NOD 小鼠子宫淋巴细胞差异表达蛋白质的蛋白质组学分析
DOI：
10.1002/jcb.22271
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发表时间：
2009-10-01
影响因子：
4
通讯作者：
Lin, Yi
中科院分区：
文献类型：
--
作者：
Li, Cui;Wang, Wenjing;Lin, Yi
与野生型（WT）小鼠相比，非肥胖糖尿病（NOD）小鼠表现出生育力受损和窝仔数减少。然而，目前还不清楚为什么异基因妊娠NOD小鼠容易发生自发胚胎丢失。在此，双向凝胶电泳（2-DE）和质谱（MS）用于检测差异表达的蛋白质在子宫淋巴细胞分离，从这些小鼠和野生型BALB/c对照。我们发现了24个差异表达蛋白。Western blot分析进一步证实了其中10个蛋白的差异表达。在24种鉴定的蛋白质中，20种在WT小鼠的子宫淋巴细胞中以比NOD小鼠高至少2倍的水平表达，而4种下调。Western blot分析证实，与NOD小鼠相比，WT小鼠子宫淋巴细胞中有8个蛋白表达上调，2个蛋白表达下调，与2-DE和MS的结果一致。此外，WT小鼠子宫淋巴细胞中高表达的蛋白在相应脾脏组中的表达水平明显降低（17/20）。这些结果表明，这些蛋白质的表达上调可能是特定的子宫淋巴细胞。报道的高表达蛋白质的功能影响怀孕期间的关键功能，包括细胞运动，细胞周期控制和代谢。用流式细胞仪分析分离的淋巴细胞中CD 3（+）和CD 49 b（+）细胞的构成比。这些差异表达的蛋白质可能参与了胚胎着床和早期发育的调控，进而影响妊娠结局。J.细胞。108：447-457，2009. (C)2009 Wiley-Liss，Inc.
Non-obese diabetic (NOD) mice exhibit impaired fertility and decreased litter size when compared to wild type (WT) mice. However, it is unclear why allogeneic pregnant NOD mice are prone to spontaneous embryo loss. Herein, two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS) were used to detect differentially expressed proteins in the uterine lymphocytes isolated from these mice and WT BALB/c controls. We found 24 differentially expressed proteins. The differential expression of 10 of these proteins was further confirmed by Western blot analysis. Out of the 24 identified proteins, 20 were expressed in uterine lymphocytes of WT mice at a level at least 2 times higher than in NOD mice, whereas 4 were down-regulated. Western blot analysis confirmed that 8 proteins were up-regulated and 2 proteins were down-regulated in WT mice compared with NOD mice, consistent with the results of 2-DE and MS. Additionally, most of the highly expressed proteins in WT uterine lymphocytes were expressed at a significantly lower level in the corresponding splenic group (17/20). These results suggest that up-regulated expression of these proteins may be specific to uterine lymphocytes. Reported functions of the highly expressed proteins affect key functions during pregnancy, including cell movement, cell cycle control, and metabolisms. Finally, we analyzed the constitutional ratio of CD3(+) and CD49b(+) cells in the isolated lymphocytes by flow cytometry. Our results suggest that the differentially expressed proteins may participate in the modulation of embryo implantation and early-stage development of embryos, and subsequently influence pregnancy outcome. J. Cell. Biochem. 108: 447-457, 2009. (C) 2009 Wiley-Liss, Inc.