PK-120表达异常在复发性流产妊娠免疫耐受机制中的作用  

Aberrant expression and possible involvement of PK-120 in immune pathogenesis of recurrent pregnancy loss

作　　者：李兰 陈燕 张东方 LI Lan;CHEN Yan;ZHANG Dongfang(Basic Medical School,North China University of Science and Technology,Tangshan 063210,China)

机构地区：[1]华北理工大学基础医学院,河北唐山063210 [2]华北理工大学附属医院妇产科

出　　处：《华北理工大学学报（医学版）》2021年第6期421-431,共11页Journal of North China University of Science and Technology：Health Sciences Edition

基　　金：河北省自然科学基金(编号:H2018209306);河北省引进留学人员资助项目(编号:C20210341);唐山市科技计划项目(编号:18130213a)。

摘　　要：目的探讨血浆激肽释放酶敏感性糖蛋白-120(PK-120)表达异常对复发性流产(RPL)妊娠免疫耐受与免疫细胞增殖的影响及其机制。方法收集2018年1月~2020年1月华北理工大学附属医院妇产科住院的60例RPL患者临床资料,同期选择30例健康女性作为对照组。Western blot法与ELISA法测定RPL患者和对照组血清、外周单核细胞(PBMC)中PK-120蛋白表达及促炎性细胞因子(Th1)、抗炎性细胞因子(Th2)含量;人CD+14单核细胞和JEG-3人绒毛滋养细胞系分别分为空白组、siControl组、siPK-120组,转染48 h后检测各组细胞中Th1、Th2的表达;分别将MOCK、PK-120(F.L.)、PK-120(△BKD)、PK-120(T.R.)表达质粒转染入JEG-3细胞,比较Th1、Th2的分泌量;CCK-8实验检测干扰及过表达的PK-120基因对免疫细胞增殖的影响;采用共培养体系分析比较Th1、Th2、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)、粘蛋白(MUC-1)、胎盘蛋白(PP-14)的表达量。结果RPL患者血清中PK-120(T.R.)裂解片段表达显著增加,对照组PK-120(F.L.)表达增加;RPL患者PBMC中PK-120 mRNA表达水平低于对照组(P<0.05),但其PK-120(T.R.)裂解片段高于对照组(P<0.01);过表达和RNA干扰实验结果显示,PK-120(F.L.)组Th2表达上调,而PK-120(T.R.)组Th1表达上调(P<0.05);PK-120 siRNA、PK-120(T.R.)可有效促进免疫细胞增殖,而PK-120(F.L.)对免疫细胞增殖有明显抑制作用(P<0.05);RPL患者血清与JEG-3细胞共培养组Th1表达显著高于对照组,且共培养后RPL患者组MMP-2、MUC-1、PP-14明显低于对照组(P<0.05)。结论PK-120(T.R.)裂解片段在RPL患者血清与PBMC中呈现高表达,且PK-120(T.R.)可明显增强炎性反应及免疫细胞增殖。Objective To explore the aberrant expression of PK-120 on immune tolerance and immune cell proliferation in recurrent pregnancy loss(RPL).Methods A total of 60 RPL patients hospitalized in the department of obstetrics and gynecology,Affiliated Hospital of North China University of Science and Technology from January 2018 to January 2020 were collected,and 30 healthy women were selected as the control group during the same period.Western blot and enzyme-linked immunosorbent assay(ELISA)were used to detect PK-120 expression and Th1/Th2 cytokine levels in normal control group and RPL patients,respectively.The tested cells were divided into control group,siRNA-negative group(siControl),and siPK-120 group.After 48 h of transfection,the expression of cytokines in each group was detected in human CD+14 monocytes and JEG-3 cells.Meantime,relative expression of inflammatory factors was measured after transfection with the MOCK,PK-120(F.L.),PK-120(△BKD),and PK-120(T.R.).CCK-8 assay was used to analyze the effect of knock-down and over-expression of PK-120 on immune cell proliferation.Co-culture with JEG-3 cells and serum derived from RPL patients or normal groups,and the relative expressions of MMP-2,MMP-9,PP-14,and MUC1 were measured.Results The relative expression of PK-120(T.R.)cleavage fragment was significantly increased,but the expression of PK-120(F.L.)was weakly expressed in serum of RPL patients.And the mRNA expression level of PK-120 in PBMCs of RPL patients was significantly lower than that of healthy control group(P<0.05);The level of PK-120(T.R.)was increased,while PK-120(F.L.)was decreased in RPL group compared with controls(P<0.01).In addition,the expression of pro-inflammatory cytokines was significantly increased after the cells were transfected with PK-120(T.R.),and anti-inflammatory cytokines were upregulated after the cells were transfected with PK-120(F.L.)(P<0.05).Moreover,knock-down of PK-120 could significantly increase immune cell proliferation,and PK-120(T.R.)could effectively promote immune cel

关 键 词：复发性流产 促炎及抗炎细胞因子 血浆激肽释放酶敏感性糖蛋白-120 

分 类 号：R714.21[医药卫生—妇产科学]