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

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英文篇名：IL-27 affects vascular endothelial cell function mediated by JAK2/STAT1 pathway in preeclampsia 作者：葛会生 ; 王寒冰 ; 黄冬妮 ; 蒋秀萍 ; 尹楠林 ; 漆洪波 英文作者：Ge Huisheng;Wang Hanbing;Huang Dongni;Jiang Xiuping;Yin Nanlin;Qi Hongbo;Department of Obstetrics and Gynecology,The First Affiliated Hospital of Chongqing Medical University; 关键词：白介素-27 ; 原代人脐静脉内皮细胞 ; JAK2/STAT1 ; 子痫前期 英文关键词：IL-27;;primary human umbilical vein endothelial cells;;JAK2/STAT1;;preeclampsia 中文刊名：ZQYK 英文刊名：Journal of Chongqing Medical University 机构：重庆医科大学附属第一医院妇产科; 出版日期：2018-05-03 16:57 出版单位：重庆医科大学学报 年：2019 期：v.44 基金：国家自然科学基金面上资助项目(编号:81471472) 语种：中文; 页：ZQYK201901001 页数：5 CN：01 ISSN：50-1046/R 分类号：7-11

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:(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:(1)Western blot was used to detect the expression of IL-27 and its receptor WSX-1 in placentas.(2)Isolation and culture of primary human umbilical vein endothelial cells(HUVECs)were conducted,and the cells were identified by immunofluorescence.(3)The 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(pJAK2)and STAT1(p-STAT1)proteins by Western blot.(4)Cell 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).(2)Primary HUVECs were successfully isolated and cultured.(3)After exposure to IL-27,JAK2/STAT1 pathway was activated in primary HUVECs.(4)The 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.

引文

[1]Sibai B,Dekker G,Kupferminc M.Pre-eclampsia[J].Lancet,2005,365(9461):785-799.
    [2]LaMarca BD,Cornelius DC,Harmon AC,et al.Identifying immune mechanisms mediating the hypertension during preeclampsia[J].Am JPhysiol Regul Integr Comp Physiol,2016,311(1):R1-9.
    [3]Awasthi A,Carrier Y,Peron JP,et al.A dominant function for interleukin 27 in generating interleukin 10-producing anti-inflammatory T cells[J].Na t Immunol,2007,8(12):1380-1389.
    [4]Hunter CA,Kastelein R.Interleukin-27:balancing protective and pathological immunity[J].Immunity,2012,37(6):960-969.
    [5]Villarino AV,Huang E,Hunter CA.Understanding the pro-and anti-inflammatory properties of IL-27[J].J Immunol,2004,173(2):715-720.
    [6]Myatt L,Webster RP.Vascular biology of preeclampsia[J].J Thromb Haemost,2009,7(3):375-384.
    [7]Tanbe AF,Khalil RA.Circulating and vascular bioactive factors during hypertension in pregnancy[J].Curr Bioact Compd,2010,6(1):60-75.
    [8]LaMarca BB,Bennett WA,Alexander BT,et al.Hypertension produced by reductions in uterine perfusion in the pregnant rat:role of tumor necrosis factor-alpha[J].Hypertension,2005,46(4):1022-1025.
    [9]Wang L,Yang T,Ding Y,et al.Chemerin plays a protective role by regulating human umbilical vein endothelial cell-induced nitric oxide signaling in preeclampsia[J].Endocrine,2015,48(1):299-308.
    [10]Yin N,Zhang H,Luo X,et al.IL-27 activates human trophoblasts to express IP-10 and IL-6:implications in the immunopathophysiology of preeclampsia[J].Mediators Inflamm,2014,2014:926875.
    [11]Xiao JP,Yin YX,Gao YF,et al.The increased maternal serum levels of IL-6 are associated with the severity and onset of preeclampsia[J].Cytokine,2012,60(3):856-860.
    [12]Leonard WJ,O'Shea JJ.Jaks and STATs:biological implications[J].Annu Rev Immunol,1998,16:293-322.
    [13]Jahantigh D,Mousavi M,Forghani F,et al.Association between maternal circulating IL-27 levels and preeclampsia[J].Cytokine,2017,102:163-167.
    [14]Pflanz S,Timans JC,Cheung J,et al.IL-27,a heterodimeric cytokine composed of EBI3 and p28 protein,induces proliferation of naive CD4+T cells[J].Immunity,2002,16(6):779-790.