反复流产患者免疫细胞谱中巨噬细胞、自然杀伤细胞和调节性T细胞的表达及其临床意义

Expression and Clinical Significance of Macrophages,Natural Killer Cell and Regulatory Cell in Immune Cell Profiles of Patients with Recurrent Miscarriage

目的通过筛选和观察免疫细胞在反复性流产(recurrent miscarriage,RM)患者中的表达及临床作用,探讨影响RM患者的潜在机制。方法筛选及下载GSE22490、GSE165004、GSE26787数据集中mRNA数据及临床数据,经Sangerbox3.0软件筛选差异基因,并获得11种免疫细胞。选取2021-01/2023-01月在作者医院就诊的52例RM患者及正常胎儿娩出且无流产病史的正常妊娠孕妇50例为研究对象。通过反转录-聚合酶链反应(reverse transcription andpolymerase chainreaction,RT-PCR)检测患者血清中调节性T细胞(regulatory cell,Treg)(FOXP3)、M1型巨噬细胞(CD86)和自然杀伤(naturalkiller,NK)细胞(CD56)的水平。结果3个数据集中筛选出165个差异基因及11种免疫细胞,其中,与正常妊娠孕妇相比,M1型巨噬细胞、NK细胞在RM患者中显著升高(P均<0.05),而Treg显著降低(P<0.05)。临床验证也提示,RM患者CD56和CD86水平较正常妊娠孕妇显著升高,而FOXP3水平显著下降(P均<0.05)。52例RM患者孕妇,流产率为9.61%,成功妊娠率为40.38%。FOXP3、CD56、CD86水平在成功妊娠孕妇与非成功妊娠孕妇中比较,差异具有统计学意义(P均<0.05)。CD86和FOXP3的异常表达可能是导致妊娠失败的原因之一(P<0.05)。结论RM孕妇可能具有独特的基因及免疫细胞,可能表现为M1型巨噬细胞、NK细胞的升高和Treg的降低。

Objective To screen and observe the expression and clinical effects of immune cells in patients with recurrent miscarriage(RM),and to explore the potential mechanism affecting RM patients.Methods The mRNA data and clinical data of GSE22490,GSE165004 and GSE26787 datasets were screened and downloaded,the differential genes were screened by Sangerbox3.0 software,and 11 kinds of immune cells were obtained.A total of 52 patients with RM and 50 patients with normal fetal delivery and no history of miscarriage were selected from January 2021 to January 2023 in author′s hospital.Serum levels of regulatory cell(Treg)(FOXP3)and M1-substyle macrophage(CD86)and natural killer(NK)cells(CD56)were examined by reverse transcription and polymerase chain reaction(RT-PCR).Results A total of 165 differential genes and 11 types of immune cells were screened in the 3 datasets,among which,compared with control group,the expressions of M1-substyle macrophage and NK cells significantly increased(all P<0.05),while the levels of Treg obviously decreased(P<0.05).Clinical validation also suggested that the levels of CD56 and CD86 were remarkably higher in patients with RM than those in normal pregnancy,however,the level of FOXP3 significantly decreased(all P<0.05).Among 52 patients with RM,the miscarriage rate was 9.61%,the successful pregnancy rate was 40.38%.The levels of FOXP3,CD56 and CD86 in pregnant women with successful pregnancy and non-successful pregnancy were compared,and the differences were statistically significant(all P<0.05).The abnormal expression of CD86 and FOXP3 might be one of the causes in pregnancy failure(P<0.05).Conclusion Pregnant women with RM may have unique genes and immune cells,which may be manifested as the increase of M1-substyle macrophages,NK cells and the decrease of Treg cells.