MK基因对小鼠子宫基质细胞蜕膜化进程的影响

Effects of Mouse Killer Gene on Decidualization of Uterine Stromal Cells in Mice

为探索肿瘤坏死因子相关凋亡诱导配体(TNF related apoptosis inducing ligand,TRAIL)的死亡受体(mouse killer,MK)对小鼠子宫基质细胞蜕膜化进程的影响,构建MK基因过表达和siRNA干扰重组腺病毒.原代培养的小鼠子宫基质细胞感染MK过表达或者干扰重组腺病毒并诱导蜕膜化,72 h后用免疫细胞化学与流式细胞术分别检测蜕膜细胞的标志物催乳素(prolactin,PRL)与蜕膜细胞凋亡率的变化情况.妊娠d4小鼠子宫角注射MK重组腺病毒,观察胚胎植入点的数量变化.实验结果表明,与对照组相比,在诱导的蜕膜细胞中过表达MK使得催乳素的含量显著降低(P<0.05),同时,蜕膜细胞的凋亡率明显升高(P<0.05),而siRNA干扰之后催乳素的含量显著升高,凋亡率明显下降(P<0.05),但是,宫角注射MK基因过表达和siRNA干扰重组腺病毒之后,胚胎植入数量均显著减少(P<0.01).提示MK基因通过参与小鼠子宫内膜基质细胞的蜕膜化进程,调节蜕膜细胞增殖与凋亡之间的平衡从而影响胚胎的植入.

To explore the effects of the death receptor（mouse killer,MK） of tumor necrosis factor-related apoptosis-inducing ligand（TRAIL） on decidualization of mouse uterine stromal cells,both of the over-expression and siRNA of MK gene recombinant adenovirus were applied to infect the primary culture of mouse uterine stromal cells followed by decidualization induction with estrogen,progesterone plus cAMP.Seventy-two hours later,immunocytochemical assay and flow cytometry were used to detect the prolactin protein expression and apoptosis of decidualized cells,which were transfected with MK recombinant adenovirus（overexpression and RNAi）.Moreover,each MK recombinant adenovirus was injected into the uterine horns of mice on the early morning of d4 of pregnancy.The number of implanting embryos was counted on d8 of pregnancy.The results showed that the levels of prolactin protein decreased significantly,but the apoptosis rate increased in the decidualized cells transfected with MK over-expression adenovirus.In the MK siRNA group,the prolatin levels were increased while the apoptosis rate was downregulated markedly.However,uterine injection of either the over-expression or the siRNA adenovirus led to dramatic decline the numbers of the implanting embryos.These results suggested that MK was involved in modulating the decidulization of mouse uterine stromal cells possibly through balancing proliferation and apoptosis of the decidualized cells.