子痫前期蜕膜基质细胞通过CX3CL1/CX3CR1途径协同dNK细胞抑制滋养细胞的增殖与侵袭  

Decidual stromal cells cooperate with dNK cells to inhibit proliferation and invasion of trophoblast through CX3CL1/CX3CR1 pathway in pre-eclampsia

作　　者：王茹 邓乾葆 张忠霞 黄素静 WANG Ru;DENG Ganbao;ZHANG Zhongxia;HUANG Sujing(Department of Obstetrics,Second Affiliated Hospital of Hainan Medical College,Haikou 570145,China)

机构地区：[1]海南医学院第二附属医院产科,海口570145

出　　处：《中国免疫学杂志》2023年第3期599-605,共7页Chinese Journal of Immunology

基　　金：海南省卫生健康行业科研项目(20A200457)。

摘　　要：目的:在子痫前期(PE)蜕膜组织中探究CX3CL1/CX3CR1的表达及其对dNK细胞功能的影响和对滋养细胞增殖与侵袭的调控。方法:取40例2018年9月至2019年10月于海南医学院第二附属医院产科住院分娩的孕产妇胎盘蜕膜组织,其中20例为PE患者(PE组),20例为正常妊娠女性(NP组),应用RT-PCR、ELISA及免疫组化(IHC)实验检测CX3CL1在两组胎盘蜕膜组织中的表达;分离早孕原代蜕膜基质细胞与蜕膜自然杀伤(dNK)细胞,细胞经形态学与流式细胞术鉴定,利用上调或沉默CX3CL1的腺病毒载体(ad-CX3CL1或ad-shCX3CL1)感染蜕膜基质细胞,RT-PCR验证感染效率;将蜕膜基质细胞与dNK细胞进行共培养,并将其分为4组:ad-vector+dNK组、ad-CX3CL1+dNK组、ad-shCX3CL1+dNK组和ad-CX3CL1+CX3CR1+dNK组;ELISA检测各组共培养体系上清液中炎症因子TNF-α与IFN-γ表达;取上述4组共培养体系的上清液处理滋养细胞系HTR8,EdU增殖实验与Transwell侵袭实验检测处理后的HTR8细胞增殖与侵袭能力的改变。结果:与NP组相比,CX3CL1在PE组胎盘蜕膜组织中的表达异常升高(P<0.01);成功分离了早孕蜕膜基质细胞与dNK细胞;与ad-vector相比,感染ad-CX3CL1或ad-shCX3CL1能显著促进或抑制蜕膜基质细胞中CX3CL1在mRNA水平的表达(P<0.01);与ad-vector+dNK组相比,ad-CX3CL1+dNK组上清液中TNF-α与IFN-γ表达明显升高(P<0.05),且共培养体系的上清液能显著抑制滋养细胞的增殖与侵袭能力(P<0.05),而ad-shCX3CL1+dNK组与ad-CX3CL1+anti-CX3CR1+dNK组上清液中TNF-α与IFN-γ表达均显著降低(P<0.05),共培养体系的上清液能显著促进滋养细胞的增殖与侵袭能力(P<0.05);且与ad-CX3CL1+dNK组相比,ad-CX3CL1+anti-CX3CR1+dNK组上清液中TNF-α与IFN-γ的表达明显降低(P<0.05),但滋养细胞的上述功能明显增加(P<0.05)。结论:CX3CL1在PE患者的胎盘蜕膜组织中的表达呈异常增高现象,且蜕膜基质细胞能够通过CX3CL1/CX3CR1途径协同dNK细胞调控滋养细胞的Objective:To investigate the expression of CX3CL1/CX3CR1 in decidua of pre-eclampsia(PE)and its effect on dNK cells and the regulation of trophoblast proliferation and invasion.Methods:The expression of CX3CL1 was detected by RT-PCR,ELISA and immunohistochemistry(IHC)in placental decidua of 40 pregnant women from September 2018 to October 2019,including 20 PE patients(PE group)and 20 normal pregnant women(NP group);decidual stromal cells and decidual natural killer(dNK)cells were isolated and identified by flow cytometry.The decidual stromal cells were infected with adenovirus vector(ad-CX3CL1 or ad-sh CX3CL1)which up-regulated or silenced CX3CL1.The infection efficiency was verified by RT-PCR;decidual stromal cells and dNK cells were co-cultured and divided into four groups:ad-vector+dNK group,ad-CX3CL1+dNK group,ad-shCX3CL1+dNK group,and ad-CX3CL1+CX3CR1+dNK group;ELISA was used to detect the expressions of inflammatory factor TNF-αand IFN-γin the supernatant of co-culture system;the supernatant of the co-culture system of the above four groups were used to treat the trophoblastic cell line HTR8.EdU proliferation test and Transwell invasion test were used to detect the changes of cell proliferation and invasion ability of the treated HTR8.Results:Compared with NP group,the expression of CX3CL1 in placental decidua of PE group was significantly higher(P<0.01);decidual stromal cells and dNK cells were isolated successfully;compared with ad-vector,infection with ad-CX3CL1 or ad-shCX3CL1 could significantly promote or inhibit the expression of CX3CL1 mRNA in decidual stromal cells(P<0.01);compared with ad-vector+dNK group,the levels of TNF-αand IFN-γin supernatant of ad-CX3CL1+dNK group were higher,while ad-shCX3CL1+dNK group,ad-CX3CL1+anti-CX3CR1+dNK group were decreased significantly(P<0.05),and the proliferation and invasion ability of trophoblast increased significantly(P<0.05);compared with ad-CX3CL1+dNK group,the levels of TNF-αand IFN-γin the supernatant of ad-CX3CL1+anti-CX3CR1+dNK group were signific

关 键 词：子痫前期 蜕膜基质细胞 CX3CL1/CX3CR1 蜕膜NK细胞 滋养细胞 增殖 侵袭 

分 类 号：R714.24[医药卫生—妇产科学]