蜕膜NK细胞协同miR-18a对人正常滋养细胞浸润能力的影响  

Influence of decidual natural killer cells with miR-18a on infiltration of human normal trophoblastic cells

作　　者：杨洋[1] 高艳 延佳佳 栾丽霞 张瑾[3] YANG Yang;GAO Yan;YAN Jia-jia;LUAN Li-xia;ZHANG Jin(Department of Gynaecology and Obstetrics,the First Affiliated Hospital of Xi'an Medical University,Xi'an,Shaanxi 710077,China)

机构地区：[1]西安医学院第一附属医院妇产科,陕西西安710077 [2]西安医学院第一附属医院超声科 [3]西安医学院第二附属医院妇产科,陕西西安710038

出　　处：《中华全科医学》2020年第2期172-176,共5页Chinese Journal of General Practice

基　　金：国家自然科学基金项目(81901510);陕西省自然科学基础研究计划项目(2018JM7115);陕西省教育厅专项科研计划项目(18JK0676)。

摘　　要：目的研究蜕膜NK细胞(decidual natural killer,dNK)能否作为母-胎界面中与滋养细胞直接接触的细胞介质,通过其分泌的相关细胞因子,协同子痫前期相关微小RNA-18a(miR-18a)影响人正常滋养细胞(HTR8)浸润能力,参与子痫前期(pre-eclapmpsia,PE)发病。方法收集2017年9月-12月于西安医学院第一附属医院妇科行人工流产患者的早孕蜕膜组织(11例),梯度密度离心联合流式细胞术分选、纯化dNK细胞;在HTR8细胞中转染miR-18a前体分子,共分3组:miR-18a过表达组、miR-18a抑制组、NC组(对照组);将dNK与上述3组HTR8细胞分别共培养;实时定量PCR(RT-qPCR)检测转染后HTR8细胞中miR-18a mRNA的表达。ELISA检测共培养24 h后各组上清液中白细胞介素8(interleukin 8,IL-8)的表达;Transwell实验分3组:共培养组、单纯培养组、对照组;检测dNK与miR-18a抑制组HTR8细胞共培养上清对滋养细胞浸润能力的影响。结果与对照组相比,miR-18a过表达和抑制均获得明显效果(均P<0.001);dNK与抑制组HTR8共培养24 h后,上清液中IL-8的表达低于对照组[(508.35±28.22)ng/L,P<0.001];与单纯培养组相比,共培养组HTR8细胞浸润能力增强(367.11±88.26,P<0.001)。结论 dNK细胞能够通过其分泌细胞因子IL-8,并协同miR-18a共同影响滋养细胞浸润能力,从而可能参与PE发病过程。Objective To study the possibility of decidual natural killer(dNK) cells as a potential cell medium in direct contact with trophoblast cells in the maternal-fetal interface through its secreted cytokines, and synergize with pre-eclampsia-associated microRNA-18 a(miR-18 a) affects the infiltration ability of human normal trophoblast cells(HTR8) and participates in the pathogenesis of pre-eclampsia(PE). Methods Of pregnant women with induced abortion from September 2017 to December 2017. Human normal early pregnancy decidual tissue(n=11) was collected, and dNK cells were sorted and purified by gradient density centrifugation combined with flow cytometry. The miR-18 a precursor molecules were transfected into HTR8 cells, which were divided into three groups: miR-18 a expression group, miR-18 a inhibition group and NC group(control group).The dNK was co-cultured with the above three groups of HTR8 cells. The real-time quantitative PCR(RT-qPCR) was used to detect the expression of miR-18 a mRNA in HTR8 cells after transfection. The expression of interleukin 8(IL-8) in the supernatant of each group was detected by ELISA after 24 hours. Transwell experiment was divided into 3 groups: co-culture group, simple culture group and control group. The effect of supernatant obtained from co-culturing of dNK and HTR8 cells on the infiltration ability of trophoblast cells in miR-18 a inhibition group were detected. Results Compared with the control group, miR-18 a overexpression and inhibitory effect was significant(all P<0.001);after co-cultured with dNK and control group HTR8 cells for 24 hours, the expression of IL-8 in the supernatant was lower than that of the control group [(508.35±28.22)ng/L, P<0.001]. Compared with the simple cultured group, the infiltration ability of HTR8 cells in co-cultured group was enhanced(367.11±88.26, P<0.001). Conclusion The dNK cells has certain influence on the infiltration of trophoblast cells through the secretion of the cytokine IL-8;and with the participation of miR-18 a, it may be invol

关 键 词：微小RNA 蜕膜NK细胞 子痫前期 滋养细胞 细胞浸润 

分 类 号：R169.42[医药卫生—公共卫生与预防医学]