IL-27通过JAK2/STAT1信号通路影响血管内皮细胞功能参与子痫前期发病机制的相关研究

IL-27 affects vascular endothelial cell function mediated by JAK2/STAT1 pathway in preeclampsia

摘要:

目的：探讨IL-27参与原代人脐静脉内皮细胞（human umbilical vein endothelial cells，HUVECs）功能受损的信号通路，及其在子痫前期发病机制中的作用。方法：①使用Western blot检测子痫前期和正常足月胎盘组织中白介素（interleukin，IL）-27及其受体WSX-1的表达；②分离并培养原代人脐静脉内皮细胞，用免疫荧光的方法鉴定HUVECs细胞；③对原代HUVECs予以50 ng/mL人重组IL-27蛋白刺激15 min、30 min、1 h和2 h后，提取蛋白，Western blot方法检测JAK2、STAT1的活化情况；④使用JAK的特异性抑制剂AG490处理原代HUVECs后，分为6个处理组，分别是空白对照组（Con）、DMSO组、抑制剂组（AG490），IL-27处理组（IL-27）、IL-27和DMSO共同处理组（DMSO+IL-27）、IL-27和抑制剂共同处理组（AG490+IL-27），然后用Transwell和细胞管腔成型实验检测IL-27对原代HUVECs血管成形能力的影响。结果：①IL-27及其受体WSX-1的蛋白水平在子痫前期胎盘组织中高于正常足月胎盘（t=2.980，P=0.020；t=2.520，P=0.040）；②成功在体外建立培养原代人脐静脉内皮细胞的模型；③人重组IL-27（50 ng/mL）刺激HUVECs细胞后可以激活JAK2/STAT1信号通路；④与相应对照组相比，IL-27组、DMSO+IL-27组的迁移和血管成形能力降低（t=16.050，P=0.000；t=16.610，P=0.000；t=11.360，P=0.000；t=11.740，P=0.000）。但是AG490+IL-27组与AG490组相比，并无明显变化（t=0.420，P=0.770；t=0.290，P=0.790）。结论：IL-27可能通过JAK2/STAT1信号通路影响血管内皮细胞功能参与子痫前期的发病。

Abstract:

Objective：To explore the mechanism of IL-27 involved in the pathogenesis of preeclampsia（PE）. Methods：①Western blot was used to detect the expression of IL-27 and its receptor WSX-1 in placentas. ②Isolation and culture of primary human umbilical vein endothelial cells（HUVECs） were conducted，and the cells were identified by immunofluorescence. ③The primary HUVECs were treated with IL-27（50 ng/mL） at time points from 0.25 to 2 hours and analyzed for activated or tyrosine phosphorylated JAK2（p-JAK2） and STAT1（p-STAT1） proteins by Western blot. ④Cell treatments of each group were as follows：normal culture group（Con），DMSO culture group，JAK2 inhibitor AG490 culture group（AG490），IL-27 culture group，DMSO+IL-27 culture group，AG490+IL-27 culture group .The tube formation assays were used to detect the effects of IL-27 on the tube formation capacity of primary HUVECs. Results：① IL-27 and WSX-1 protein levels were both increased in the PE group compared with those of the control group（t=2.98０，P=0.02０；t=2.52０，P=0.04０）. ②Primary HUVECs were successfully isolated and cultured. ③After exposure to IL-27，JAK2/STAT1 pathway was activated in primary HUVECs. ④The migration and tube formation abilities of IL-27 group and DMSO+IL-27 group were significantly reduced（t=16.05０，P=0.00０；t=16.61０，P=0.00０；t=11.36０，P=0.00０；t=11.74０，P=0.00０）. There was no significantly difference between AG490 group and AG490+IL-27 group（t=0.420，P=0.770；t=0.290，P=0.790）. Conclusion：IL-27 may play a critical role in the pathogenesis of PE through JAK2/STAT1 pathway.