PGRN对胚胎植入及胎盘血管形成的影响

Effect of progranulin on embryo implantation and placental angiogenesis

目的:探讨颗粒前体蛋白(progranulin,PGRN)对小鼠胎盘血管形成的影响。方法:收集2017年1至4月在重庆医科大学附属第一医院行剖宫产手术的子痫前期(pre-eclampsia,PE)组(10例)以及正常对照(normal control,NC)组(12例)的胎盘组织;通过PCR及Western blot比较子痫前期组和正常对照组胎盘组织中PGRN蛋白的表达情况;通过免疫荧光以及免疫组化等方式来对比在孕6.5 d胚胎及孕17.5 d胎盘在PGRN-/-小鼠和正常小鼠血管生成情况;苏木素-伊红染色比较孕6.5 d子宫动脉管径大小及管壁厚度。结果:PGRN在子痫前期组胎盘组织中的表达明显高于正常对照组,差异有统计学意义(1.420±0.255,0.726±0.192,t=4.118,P=0.001);孕6.5 d时,PGRN-/-小鼠的胚胎个数较正常对照明显减少(5.333±1.154,9.333±1.527,t=3.618,P=0.022);在孕早期血管标志物SMA蛋白在PGRN-/-小鼠表达较对照组减弱(25 396.600±2 413.606,17 484.980±1 606.019,t=4.727,P=0.009);PGRN-/-小鼠子宫动脉较正常小鼠子宫动脉直径稍减小,血管肌层厚度减少较明显(675.844±147.635,1 022.234±133.902,t=3.010,P=0.039;10.136±1.634,18.214±2.899,t=4.204,P=0.013);孕17.5 d,血管标志物SMA、COX2及Tie2蛋白在PGRN-/-小鼠胎盘迷路层表达量比正常对照组减弱(626.467±114.744,5 288.375±871.319,t=9.188,P=0.001;828.298±29.192,1169.734±43.490,t=11.290,P=0.000;335.766±125.570,2 415.752±579.038,t=6.080,P=0.003)。结论:PGRN可能通过参与胚胎和胎盘的血管形成,在妊娠相关并发症的发生中起作用。

Objective:To investigate the effect of progranulin(PGRN)on placental angiogenesis in mice. Methods:Placenta tissue samples were collected from 10 patients with preeclampsia(PE)who underwent caesarean section in The First Affiliated Hospital of Chongqing Medical University from January to April,2017(PE group) and 12 normal controls(NC group). PCR and Western blot were used to compare the expression of PGRN in the placenta between the PE group and the NC group. Immunofluorescence assay and immunohistochemistry were used to compare angiogenesis in the embryo on day 6.5 of pregnancy and in the placenta on day 17.5 of pregnancy between PGRN-/-mice and normal mice. HE staining was used to compare the diameter and wall thickness of the uterine artery on day 6.5 of pregnancy between groups. Results:The PE group had significantly higher PGRN expression in placenta than the NC group(1.420±0.255 vs. 0.726±0.192,t=4.118,P=0.001). On day 6.5 of pregnancy,the PGRN-/-mice had a significantly lower number of embryos than the normal mice(5.333±1.154 vs. 9.333±1.527,t=3.618,P=0.022). The PGRN-/-mice had significantly lower expression of the vascular marker SMA protein than the normal mice in early pregnancy(25 396.600±2 413.606 vs. 17 484.980±1 606.019,t=4.727,P=0.009). Compared with the normal mice,the PGRN-/-mice had significant reductions in the diameter of the uterine artery(675.844±147.635 vs. 1 022.234±133.902,t=3.010,P=0.039)and muscular thickness of the uterine artery(10.136±1.634 vs. 18.214±2.899,t=4.204,P=0.013). On day 17.5 of pregnancy,the PGRN-/-mice had significantly lower expression of the vascular markers SMA,COX2,and Tie2 than the normal mice(SMA:626.467±114.744 vs. 5 288.375±871.319,t=9.188,P=0.001;COX2:828.298±29.192 vs. 1169.734±43.490,t=11.290,P=0.000;Tie2:335.766±125.570 vs. 2 415.752±579.038,t=6.080,P=0.003). Conclusion:PGRN may play a role in the development of pregnancy-related complications by participating in angiogenesis of the embryo and the placenta.