Yu Nan, Yan Wenjie, Yin Tailang, Wang Yaqin, Guo Yue, Zhou Danni, Xu Mei, Ding Jinli, Yang Jing
Reproductive Medical Center, Renmin Hospital of Wuhan University, Wuhan, People's Republic of China; Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, People's Republic of China.
Reproductive Medical Center, Renmin Hospital of Wuhan University, Wuhan, People's Republic of China.
PLoS One. 2015 Jun 18;10(6):e0125589. doi: 10.1371/journal.pone.0125589. eCollection 2015.
Successful embryo implantation and placentation depend on appropriate trophoblast invasion into the maternal endometrial stroma. Human chorionic gonadotropin (hCG) is one of the earliest embryo-derived secreted signals in the peripheral blood mononuclear cells (PBMC) that abundantly expresses hCG receptors. The aims of this study were to estimate the effect of human embryo-secreted hCG on PBMC function and investigate the role and underlying mechanisms of activated PBMC in trophoblast invasion. Blood samples were collected from women undergoing benign gynecological surgery during the mid-secretory phase. PBMC were isolated and stimulated with or without hCG for 0 or 24 h. Interleukin-1β (IL-1β) and leukemia inhibitory factor (LIF) expressions in PBMC were detected by enzyme-linked immunosorbent assay and real-time polymerase chain reaction (PCR). The JAR cell line served as a model for trophoblast cells and was divided into four groups: control, hCG only, PBMC only, and PBMC with hCG. JAR cell invasive and proliferative abilities were detected by trans-well and CCK8 assays and matrix metalloproteinase (MMP)-2 (MMP-2), MMP-9, vascular endothelial growth factor (VEGF), tissue inhibitor of metalloproteinase (TIMP)-1, and TIMP-2 expressions in JAR cells were detected by western blotting and real-time PCR analysis. We found that hCG can remarkably promote IL-1β and LIF promotion in PBMC after 24-h culture. PBMC activated by hCG significantly increased the number of invasive JAR cells in an invasion assay without affecting proliferation, and hCG-activated PBMC significantly increased MMP-2, MMP-9, and VEGF and decreased TIMP-1 and TIMP-2 expressions in JAR cells in a dose-dependent manner. This study demonstrated that hCG stimulates cytokine secretion in human PBMC and could stimulate trophoblast invasion.
成功的胚胎着床和胎盘形成依赖于滋养层细胞对母体子宫内膜基质的适当侵入。人绒毛膜促性腺激素（hCG）是外周血单个核细胞（PBMC）中最早由胚胎分泌的信号之一，PBMC中大量表达hCG受体。本研究的目的是评估人胚胎分泌的hCG对PBMC功能的影响，并研究活化的PBMC在滋养层细胞侵入中的作用及潜在机制。在分泌中期从接受良性妇科手术的女性中采集血样。分离PBMC，用或不用hCG刺激0或24小时。通过酶联免疫吸附测定和实时聚合酶链反应（PCR）检测PBMC中白细胞介素-1β（IL-1β）和白血病抑制因子（LIF）的表达。JAR细胞系用作滋养层细胞模型，分为四组：对照组、仅hCG组、仅PBMC组和hCG与PBMC组。通过Transwell和CCK8测定检测JAR细胞的侵袭和增殖能力，通过蛋白质印迹和实时PCR分析检测JAR细胞中基质金属蛋白酶（MMP）-2、MMP-9、血管内皮生长因子（VEGF）、金属蛋白酶组织抑制剂（TIMP）-1和TIMP-2的表达。我们发现，培养24小时后，hCG可显著促进PBMC中IL-1β和LIF的分泌。hCG激活的PBMC在侵袭试验中显著增加了JAR细胞的侵袭数量，但不影响其增殖，并且hCG激活的PBMC以剂量依赖的方式显著增加JAR细胞中MMP-2、MMP-9和VEGF的表达，并降低TIMP-1和TIMP-（此处原文有误，应为TIMP-2）的表达。本研究表明，hCG刺激人PBMC中的细胞因子分泌，并可刺激滋养层细胞侵入。
