妊娠对类风湿关节炎患者内皮祖细胞的影响及机制研究

Effect of pregnancy on endothelial progenitor cells in patients with rheumatoid arthritis and its mechanism

目的:探讨妊娠对RA患者内皮祖细胞(EPC)的影响及机制。方法:将2016年1月至2018年6月来本院就诊的RA初治患者纳入本项研究,按照有无妊娠,将患者分为单纯RA组和RA合并妊娠组,均为女性患者,每组各30例;免疫组织化学染色检测滑膜组织内淋巴细胞共同抗原(LCA)+淋巴细胞和CD68+巨噬细胞的数目;流式细胞术检测EPC和内皮细胞的比例;ELISA检测EPC上清中血管内皮生长因子(VEGF)、基质细胞衍生因子(SDF)-1、IL-6和IL-10的浓度。2组之间比较采用t检验,多组间比较用单因素方差分析。结果:免疫组织化学结果显示RA合并妊娠组关节滑膜内CD68+巨噬细胞和LCA+淋巴细胞的数目低于单纯RA组;ELISA的结果显示单纯RA组外周血可溶性人HLA-G的浓度为(8.9±1.7)pg/ml,RA合并妊娠组的浓度为(396.7±89.6)pg/ml,2组差异有统计学意义(t=4.329,P<0.01);流式细胞学检测结果显示单纯RA组患者淋巴细胞中EPC的比例为(0.13±0.03)%,RA合并妊娠组的比例为(0.76±0.09)%,2组差异有统计学意义(t=6.671,P<0.01);相关性分析结果显示2组患者外周血中EPC比例与可溶性HLA-G浓度呈正相关(r=0.8861,P<0.01);体外的实验结果显示HLA-G能够促进单纯RA患者EPC旁分泌和分化功能的恢复。结论:妊娠能够提高RA患者体内EPC的数量和生物学功能,HLA-G在这个过程中可能发挥着重要作用。

Objective To investigate the effect of pregnancy on endothelial progenitor cells(EPCs)in patients with rheumatoid arthritis(RA)and its mechanism.Methods The newly treated RA patients in our hospital from January 2016 to June 2018,were included in this study.According to pregnancy or not,patients were divided into simple RA group and RA pregnancy group.They were all female patients,30 in each group.Immunohistochemical staining was used to detect the number of lymphocyte common antigen(LCA)+lymphocytes and CD68+macrophages in synovial tissue,flow cytometry was used to detect the proportion of EPC and endothelial cells,and enzyme-linked immunosorbent assay(ELISA)was used to detect the concentrations of vascular endothelial growth factor(VEGF),stromal cell derived factor(SDF-1),interleukin(IL)-6 and IL-10 in EPC supernatant.T-test was used for the comprarison between the two groups,and single factor analysis of variancewas used for the comparison between multiple groups.Results Immunohistoche-mical results showed that the number of CD68+macrophages and LCA+lymphocytes in synovium of RA with pregnancy group was significantly lower than that of non-pregnant RA group.The results of ELISA showed that the concentration of human leucocyte antigen-G(HLA-G)in peripheral blood was(8.9±1.7)pg/ml in non-pregnant RA group and(396.7±89.6)pg/ml in RA pregnancy group,the difference beween the two groups was statistically significant(t=4.329,P<0.01).The results of flow cytometry showed that the proportion of EPC in lymphocytes was(0.13±0.03)%in non-pregnant RA group and(0.76±0.09)%in RA with pregnancy group,the difference beween the two groups was statisti-cally significant(t=6.671,P<0.01).The results of correlation analysis showed that the proportion of EPC in peripheral blood was positively correlated with HLA-G concentration(r=0.8861,P<0.01).In vitro experiments showed that HLA-G could promote the recovery of EPC paracrine and differentiation function in RA patients.Conclusion Pregnancy can improve the number and biological function of EPC in patients with RA.HLA-G may play an important role in this process.