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

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:(1)IL-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。

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.