Objective To study the role of IFN-γ/IL-10 cytokines protein expression of human decidual stromal cells（DSC） vitro. on IL-10 receptor gene and in human early pregnancy in vitro. Methods Human DSC was isolated and c...Objective To study the role of IFN-γ/IL-10 cytokines protein expression of human decidual stromal cells（DSC） vitro. on IL-10 receptor gene and in human early pregnancy in vitro. Methods Human DSC was isolated and cultured in vitro, and the expression of IL-10R1 and IL-10R2 gene was analyzed after cells had been treated with TH2-type cytokines IL-10 and TH1-type cytokines IFN-γ within 60 rain with semiquantitative reverse transcriptase-PCR, then the influence of IL-10 and IFN-γ on expression of IL-10R protein was examined by first trimester DSC using flow cytometry. In addition, the vitality of DSC was detected by MTT. Results IL-10R1 mRNA levels of DSC treated with IL-10 （10 ng/ml） reached the peak level within 15 rain, and were significantly lower at 30 rain, then were not detected at 45 min. The expression of IL-10R1 were induced to moderate level by IFN-γ（10 ng/ml） within 30 rain, and reduced to undetected levels at 60 min. There was no significant difference of IL-10R2 expression （P〉0.05） between treated and not with the abovementioned cytokines. The IL-10R protein expression and vitality of DSC were significantly enhanced by IL-10 （10 ng/ml） and IFN-γ （10 ng/ml） which treated DSC 48 h （P〈0.05）. Coneclusion IL-10 and IFN-γ may play an important role of biologic function in early pregnancy by influencing IL-10R expression of DSC.展开更多
Summary: An early embryo co-culture system with human decidual stromal cells was established to study its effect on early embryonic cleavage and growth in vitro. Three hundred and eight 2-cell mouse embryos were co-cu...Summary: An early embryo co-culture system with human decidual stromal cells was established to study its effect on early embryonic cleavage and growth in vitro. Three hundred and eight 2-cell mouse embryos were co-cultured with human decidual stromal cell monolayer in MEM+0. 4 % bovine serum albumin (BSA) and 163 embryos cultured in MEM+15 % FCS alone as control. Among the mouse 2-cell embryos co-cultured with human decidual stromal cells, 72.73 % developed to the morula stage and 67.21 % cavitated to blastocysts with 59. 74 % hatching, as compared with 61. 34 % to morula stage, 48. 47 % to blastocysts and none hatching in the controls, respectively. Co-cultured embryos cleaved slightly faster than controls and showed no or less fragmentation than those in the control. These results suggested that human decidual stromal cells can support early embryonic development and yield a reasonable number of embryos with good quality up to blastocyst stage.展开更多
Endometrial stromal cell decidualization is a crucial step in endometrial remodeling during pregnancy.Decidualization is controlled by orchestrated ovarian hormones,followed by the activation of various downstream sig...Endometrial stromal cell decidualization is a crucial step in endometrial remodeling during pregnancy.Decidualization is controlled by orchestrated ovarian hormones,followed by the activation of various downstream signaling pathways.Accumulating evidence has shown multiple functions of decidualized endometrial stromal cells during embryo implantation,including tissue remodeling,antioxidative stress,angiogenesis,and immune tolerance.The distinct secretomes of decidualized stromal cells also reveal their intensive interactions with epithelial,endothelial,and immune cells.However,aberrant decidualization leads to pregnancy failures,such as recurrent pregnancy loss and repeated implantation failure.This review aimed to provide an overview of the molecular mechanisms underlying the divergent functions of decidualized endometrial stromal cells and their potential clinical applications.Moreover,the use of single-cell RNA sequencing data further enhances our understanding of these biological processes.This review discusses decidualization-related signaling pathways that serve as potential therapeutic targets for treating implantation failure in in vitro fertilization and provides novel approaches to investigate the underlying causes of female infertility.展开更多
T helper 17 （Th17） cells have both regulatory and protective roles in physiological conditions. The Th17 subset and the cytokine interleukin-17A （IL-17A） have been implicated in the pathogenesis of certain autoimm...T helper 17 （Th17） cells have both regulatory and protective roles in physiological conditions. The Th17 subset and the cytokine interleukin-17A （IL-17A） have been implicated in the pathogenesis of certain autoimmune diseases, several types of cancer and allograft rejection. However, the role of Th17 cells at the maternal/fetal interface remains unknown. Here, we demonstrate that Th17 cells are present in decidua and are increased in the peripheral blood of 10 clinically normal pregnancies based on intracellular cytokine analysis. Our results suggest a potential role of Th17 cells in sustaining pregnancy in humans. Furthermore, we demonstrate that decidual stromal cells （DSCs） but not trophoblast cells recruit peripheral Th17 cells into the decidua by secreting CCL2. The recruited Th17 cells promote proliferation and invasion and inhibit the apoptosis of human trophoblast cells by secreting IL-17 during the first trimester of pregnancy. These findings indicate a novel role for Th17 cells in controlling the maternal-fetal relationship and placenta development.展开更多
目的:研究正常早孕绒毛及蜕膜组织细胞因子信号转导负调控因子(Suppressors of cytok ine signaling,SOCS)基因和蛋白水平表达,以揭示SOCS在母胎界面生理性调节作用。方法:半定量RT-PCR检测早孕绒毛组织、蜕膜组织及原代培养早孕滋养细...目的:研究正常早孕绒毛及蜕膜组织细胞因子信号转导负调控因子(Suppressors of cytok ine signaling,SOCS)基因和蛋白水平表达,以揭示SOCS在母胎界面生理性调节作用。方法:半定量RT-PCR检测早孕绒毛组织、蜕膜组织及原代培养早孕滋养细胞、蜕膜基质细胞SOCS1、SOCS2、SOCS3mRNA水平;W estern b lot检测早孕绒毛组织及蜕膜组织SOCS1、SOCS2、SOCS3蛋白表达;免疫组化定位SOCS1、SOCS2、SOCS3在早孕绒毛组织、蜕膜组织表达;ELISA检测滋养细胞、蜕膜基质细胞分泌IL-10、IFNγ-。结果:正常母胎界面见SOCS1、SOCS2、SOCS3基因表达,其中SOCS3绒毛/蜕膜阳性率73.7%/71.1%;SOCS2绒毛/蜕膜阳性率50.0%/39.5%,SOCS1最少,绒毛/蜕膜阳性率34.2%/31.6%;SOCS1、SOCS2、SOCS3蛋白表达与转录水平基本一致;正常母胎界面SOCS1、SOCS2、SOCS3表达主要定位于绒毛滋养细胞和蜕膜间质;体外无血清培养滋养细胞和蜕膜基质细胞SOCS2、SOCS3低表达,SOCS1未见表达,其分泌的IL-10随时间而增高(P<0.05)。结论:正常早孕母胎界面表达SOCS1、SOCS2、SOCS3,无刺激条件下滋养细胞和蜕膜基质细胞低表达SOCS2、SOCS3,SOCS在正常妊娠Th平衡中具有重要意义。展开更多
基金This study was supported by research grant from National Natural Science Foundation of China (No.30572446), research grants from Modern Biology & Pharmacy Foundation of ShanghaiScience Committee (No.02D219115) and Fudan University (985 Program).
文摘Objective To study the role of IFN-γ/IL-10 cytokines protein expression of human decidual stromal cells（DSC） vitro. on IL-10 receptor gene and in human early pregnancy in vitro. Methods Human DSC was isolated and cultured in vitro, and the expression of IL-10R1 and IL-10R2 gene was analyzed after cells had been treated with TH2-type cytokines IL-10 and TH1-type cytokines IFN-γ within 60 rain with semiquantitative reverse transcriptase-PCR, then the influence of IL-10 and IFN-γ on expression of IL-10R protein was examined by first trimester DSC using flow cytometry. In addition, the vitality of DSC was detected by MTT. Results IL-10R1 mRNA levels of DSC treated with IL-10 （10 ng/ml） reached the peak level within 15 rain, and were significantly lower at 30 rain, then were not detected at 45 min. The expression of IL-10R1 were induced to moderate level by IFN-γ（10 ng/ml） within 30 rain, and reduced to undetected levels at 60 min. There was no significant difference of IL-10R2 expression （P〉0.05） between treated and not with the abovementioned cytokines. The IL-10R protein expression and vitality of DSC were significantly enhanced by IL-10 （10 ng/ml） and IFN-γ （10 ng/ml） which treated DSC 48 h （P〈0.05）. Coneclusion IL-10 and IFN-γ may play an important role of biologic function in early pregnancy by influencing IL-10R expression of DSC.
文摘Summary: An early embryo co-culture system with human decidual stromal cells was established to study its effect on early embryonic cleavage and growth in vitro. Three hundred and eight 2-cell mouse embryos were co-cultured with human decidual stromal cell monolayer in MEM+0. 4 % bovine serum albumin (BSA) and 163 embryos cultured in MEM+15 % FCS alone as control. Among the mouse 2-cell embryos co-cultured with human decidual stromal cells, 72.73 % developed to the morula stage and 67.21 % cavitated to blastocysts with 59. 74 % hatching, as compared with 61. 34 % to morula stage, 48. 47 % to blastocysts and none hatching in the controls, respectively. Co-cultured embryos cleaved slightly faster than controls and showed no or less fragmentation than those in the control. These results suggested that human decidual stromal cells can support early embryonic development and yield a reasonable number of embryos with good quality up to blastocyst stage.
基金supported by the RGC General Research Fund(17120720 to K.F.L.)the Professor PC Ho Research and Development Fund in Reproductive Medicine from the Department of Obstetrics and Gynecology,University of Hong Kong and Health and Medical Research Fund,Hong Kong(L.H.J.)a Conference and Research Committee grant from the University of Hong Kong(K.F.L.).
文摘Endometrial stromal cell decidualization is a crucial step in endometrial remodeling during pregnancy.Decidualization is controlled by orchestrated ovarian hormones,followed by the activation of various downstream signaling pathways.Accumulating evidence has shown multiple functions of decidualized endometrial stromal cells during embryo implantation,including tissue remodeling,antioxidative stress,angiogenesis,and immune tolerance.The distinct secretomes of decidualized stromal cells also reveal their intensive interactions with epithelial,endothelial,and immune cells.However,aberrant decidualization leads to pregnancy failures,such as recurrent pregnancy loss and repeated implantation failure.This review aimed to provide an overview of the molecular mechanisms underlying the divergent functions of decidualized endometrial stromal cells and their potential clinical applications.Moreover,the use of single-cell RNA sequencing data further enhances our understanding of these biological processes.This review discusses decidualization-related signaling pathways that serve as potential therapeutic targets for treating implantation failure in in vitro fertilization and provides novel approaches to investigate the underlying causes of female infertility.
文摘T helper 17 （Th17） cells have both regulatory and protective roles in physiological conditions. The Th17 subset and the cytokine interleukin-17A （IL-17A） have been implicated in the pathogenesis of certain autoimmune diseases, several types of cancer and allograft rejection. However, the role of Th17 cells at the maternal/fetal interface remains unknown. Here, we demonstrate that Th17 cells are present in decidua and are increased in the peripheral blood of 10 clinically normal pregnancies based on intracellular cytokine analysis. Our results suggest a potential role of Th17 cells in sustaining pregnancy in humans. Furthermore, we demonstrate that decidual stromal cells （DSCs） but not trophoblast cells recruit peripheral Th17 cells into the decidua by secreting CCL2. The recruited Th17 cells promote proliferation and invasion and inhibit the apoptosis of human trophoblast cells by secreting IL-17 during the first trimester of pregnancy. These findings indicate a novel role for Th17 cells in controlling the maternal-fetal relationship and placenta development.
文摘目的:研究正常早孕绒毛及蜕膜组织细胞因子信号转导负调控因子(Suppressors of cytok ine signaling,SOCS)基因和蛋白水平表达,以揭示SOCS在母胎界面生理性调节作用。方法:半定量RT-PCR检测早孕绒毛组织、蜕膜组织及原代培养早孕滋养细胞、蜕膜基质细胞SOCS1、SOCS2、SOCS3mRNA水平;W estern b lot检测早孕绒毛组织及蜕膜组织SOCS1、SOCS2、SOCS3蛋白表达;免疫组化定位SOCS1、SOCS2、SOCS3在早孕绒毛组织、蜕膜组织表达;ELISA检测滋养细胞、蜕膜基质细胞分泌IL-10、IFNγ-。结果:正常母胎界面见SOCS1、SOCS2、SOCS3基因表达,其中SOCS3绒毛/蜕膜阳性率73.7%/71.1%;SOCS2绒毛/蜕膜阳性率50.0%/39.5%,SOCS1最少,绒毛/蜕膜阳性率34.2%/31.6%;SOCS1、SOCS2、SOCS3蛋白表达与转录水平基本一致;正常母胎界面SOCS1、SOCS2、SOCS3表达主要定位于绒毛滋养细胞和蜕膜间质;体外无血清培养滋养细胞和蜕膜基质细胞SOCS2、SOCS3低表达,SOCS1未见表达,其分泌的IL-10随时间而增高(P<0.05)。结论:正常早孕母胎界面表达SOCS1、SOCS2、SOCS3,无刺激条件下滋养细胞和蜕膜基质细胞低表达SOCS2、SOCS3,SOCS在正常妊娠Th平衡中具有重要意义。
文摘目的·探讨不明原因复发性流产(unexplained recurrent pregnancy loss,URPL)和正常妊娠早期,泌乳素(prolactin,PRL在外周血和蜕膜基质细胞(decidual stromal cells,DSCs)中的表达差异。方法·纳入2018年3月—2019年3月于上海交通大学医学院附属仁济医院妇产科就诊的80例URPL患者、70例人工流产妇女以及190例正常妊娠妇女。采用酶联免疫吸附方法检测外周血PRL水平。采用实时定量聚合酶链式反应(quantitative real time PCR,RT-qPCR)、蛋白质免疫印迹(Western blotting,WB)及免疫组化染色(immunohistochemistry,IHC)方法检测DSCs中PRL的mRNA和蛋白表达水平。2组之间比较采用独立样本t检验,多组组间比较采用单因素方差分析。结果·排除21例胚胎染色体异常,UPRL组最终纳入59例。人工流产组纳入70例。190例正常妊娠妇女中177例完成随访,其中157例活产,被归于正常妊娠活产组,其余20例发生胚胎停育或自然流产,其中流产发生于孕10周以前的19例被归入正常妊娠流产组。正常妊娠活产组和人工流产组妇女的外周血PRL水平类似,相同孕周间差异无统计学意义,且都随着孕周增长显著升高(正常妊娠活产组:孕5-5^(+6)周vs孕6~7^(+6)周,P=0.002;孕6~7^(+6)周vs孕8~9^(+6)周,P=0.012。人工流产组:孕5-5^(+6)周vs孕6~7^(+6)周,P=0.015,孕6~7^(+6)周vs孕8~9^(+6)周,P=0.023);URPL患者的外周血PRL水平随着孕周增长无显著变化,并且从孕6周起显著低于相同孕周的正常妊娠活产妇女和人工流产妇女(孕6~7^(+6)周:P=0.018,0.024;孕8~9^(+6)周:P=0.015,0.003);正常妊娠流产组的外周血PRL水平也与相同孕周的URPL组患者类似。人工流产组妇女DSCs的PRL表达在mRNA和蛋白水平上都随着孕周增长显著升高(孕5-5^(+6)周vs孕6~7^(+6)周:RT-qPCR,P=0.020;WB,P=0.010;IHC,P=0.030。孕6~7^(+6)周vs孕8~9^(+6)周:RT-qPCR,P=0.011;WB,P=0.034;IHC,P=0.012);URPL组DSCs的PRL表达水平增长迟滞:孕6~7^(+6)周及孕8~9^(+6)周的URPL妇女DSCs的PRL表达水平显著低于相同孕周的人工流产妇女(孕6~7^(+6)周:RT-qPCR,P=0.012;WB,P=0.014;IHC,P=0.028;孕8~9^(+6)周:RT-qPCR,P=0.011;WB,P=0.030;IHC,P=0.026)。结论·UPRL患者DSCs的PRL表达缺陷,DSCs蜕膜化不足可能与URPL的发生有关;孕6周以后外周血PRL低水平可能预示胚胎发育不良。
