PD-1/PD-L1信号通路调控Treg/Th17在子痫前期发生发展中的机制研究

Mechanism of PD-1/PD-L1 signaling pathway regulating Treg/Th17 in the occurrence and development of preeclampsia

目的:通过检测子痫前期(PE)及正常孕妇胎盘中程序性细胞死亡受体1(programmed cell death1,PD-1)及其配体PD-L1、调节性细胞和辅助性T细胞的特异性核转录因子叉头状螺旋转录因子3(forkhead box protein 3,Foxp3)和维甲酸相关孤核受体γc(retinoci acid receptor-related orphan receptoe gammat,RORγc)的表达水平,研究它们的差异表达与PE发病的关系。方法:选取2021年3月~2022年3月在海南医学院附属海南医院产科接受诊治的子痫前期患者40例,分为轻度PE组(PE组,n=20)和重度PE组(sPE组,n=20),另取同期剖宫产终止妊娠的正常孕妇作为对照组(对照组,n=20)。收集3组孕妇剖宫产后的胎盘组织,采用实时荧光定量PCR检测胎盘PD-1、PD-L1、Foxp3和RORγc的mRNA表达,而蛋白表达情况采用蛋白质印迹法检测,并分析其与PE发生发展的关系。结果:(1)qRT-PCR法检测结果表明,与对照组相比,PE组和sPE组的PD-1、PD-L1、Foxp3的mRNA表达均显著下降;与PE组相比,sPE组患者3项指标的mRNA表达水平下降的更显著,3组数据两两比较差异均有统计学意义(P<0.05)。与对照组相比,PE组和sPE组RORγc的mRNA表达均显著上升;与PE组相比,sPE组患者RORγc的mRNA表达水平升高更显著,3组数据两两比较差异均有统计学意义(P<0.05)。(2)蛋白质印迹法结果显示,与对照组相比,PE组和sPE组的PD-1、PD-L1、Foxp3蛋白表达水平明显下降;与PE组相比,sPE组患者3项指标的蛋白表达水平下降更显著,3组数据两两比较差异均有统计学意义(P<0.05)。与对照组相比,PE组和sPE组RORγc的蛋白表达水平明显上升;与PE组相比,sPE组患者RORγc的蛋白表达水平升高更显著,3组数据两两比较差异均有统计学意义(P<0.05)。结论:PD-1/PD-L1异常表达通过引起调节性T细胞/辅助性T细胞-17免疫失衡参与子痫前期的发生及进展。

Objective:To detect the expression levels of programmed cell death 1(PD-1)and its ligand PD-L1,regulatory cell(Treg)and T helper cell 17(Th17)specific nuclear transcription factors forkhead box protein 3(Foxp3)and the retinoic acid-related orphan nuclear receptorgammat(RORγc)in the placenta of normal pregnant women and in preeclamptic(PE)women,and to study the relationship between their differential expression and the development of PE.Methods:Between March 2021and March 2022,40 patients with pre-eclampsia who were treated at the Obstetrical Department in Hainan Hospital were selected,including 20 cases of mild(PE)and 20 cases of severe(sPE),and 20 normal pregnant women with singleton cesarean section in the same period were selected to collect placental tissue.PD-1,PD-L1,Foxp3 and RORγc in placenta were detected by qRT-PCR and Western blot.Results:(1)The results of qRT-PCR assay showed that the mRNA expression of PD-1,PD-L1,Foxp3 protein decreased significantly compared with the control group.The changes in PD-1,PD-L1 and Foxp3 became more obvious as the disease progressed(P<0.05);The mRNA expression of RORγc increased significantly compared with the control group.The changes of RORγc mRNA became more obviousas as the disease progressed(P<0.05);(2)The results of Western blot assay show,compared with the normal group,the protein expression of PD-1,PD-L1,Foxp3 mRNA decreased significantly.With the aggravation of the disease,the changes of PD-1 and PD-L1 became more obvious,with significant difference(P<0.05);Compared with the normal group,the protein expression of RORγc increased significantly.With the aggravation of the disease,the changes of RORγc protein became more obvious with significant difference(P<0.05).Conclusion:PD-1/PD-Ll participates in the occurrence and development of preeclampsia by regulating Treg/Th17 immune balance.