人早孕期滋养细胞通过表达趋化因子SDF-1募集蜕膜免疫活性细胞

目的　研究人早孕期绒毛及滋养细胞趋化因子SDF 1的表达、分泌及其对蜕膜免疫活性细胞的募集作用。方法　收集早孕期绒毛组织并检测SDF 1的定位表达 ;用ELISA分析早孕期滋养细胞培养上清液中SDF 1的浓度水平 ;分离早孕期蜕膜免疫活性细胞 ,以趋化试验研究SDF 1及滋养细胞培养上清液对蜕膜免疫活性细胞的趋化作用。结果　人早孕期滋养细胞表达并分泌SDF 1;一定浓度范围的SDF 1对蜕膜免疫活性细胞具有趋化作用 ,最大趋化指数为 3 .2 7± 0 .89;滋养细胞培养上清液亦明显趋化蜕膜免疫活性细胞 ,趋化指数为 2 .11± 0 .79。结论　人早孕期滋养细胞表达的SDF 1对蜕膜免疫活性细胞具有趋化、募集作用 ,从而有助于形成蜕膜独特的淋巴细胞群体并维持母 胎免疫耐受     

早孕期人滋养细胞CXCR4/CXCL12的表达及低氧对其的影响

目的：研究人早孕期滋养细胞CXCR4/CXCL12的表达情况及低氧对其表达的调节作用.方法：免疫组织化学法检测早孕期绒毛组织中及原代培养的滋养细胞CXCR4/CXCL12的表达情况;实时定量PCR检测低氧状态下滋养细胞CXCR4 mRNA水平.结果：胎盘绒毛柱、滋养细胞及血管内均有CXCR4/CXCL12表达;低氧培养12、24、48和72小时后滋养细胞CXCR4 mRNA表达增加,显著高于正常氧条件下的表达情况.结论：早孕期胎盘表达CXCR4/CXCL12对于维系正常妊娠的顺利进行可能发挥重要作用;低氧是CXCR4表达的重要调节因子,可能参与了妊娠的病理生理过程.     

流式细胞仪分离人早孕绒毛外滋养细胞及鉴定

目的用流式细胞仪分离早期绒毛外滋养细胞，以便对其功能进行进一步研究。方法用胰蛋白酶消化法得到早期绒毛细胞，用流式细胞仪分离纯化绒毛外滋养细胞。所得的细胞用免疫细胞化学，光镜，透射电镜检以及Westen-blot方法鉴定。结果使用流式细胞仪成功的分离了绒毛外滋养细胞，纯度超过98％。结论此方法可快速准确大量获得绒毛外滋养细胞。     

早孕绒毛滋养细胞FasL表达异常与自然流产的关系

通过比较正常早孕与自然流产者滋养细胞表面FasL表达,进一步从分子免疫学角度探讨自然流产的发病机理。方法：用特异性的FasL抗体进行免疫组化染色,并通过高清晰彩色病理免疫组化测量系统对其定量分析,SSPS对两组进行比较。结果：自然流产组滋养细胞表面FasL表达面积及强度均明显低于正常早孕组,两者间差异有显著性意义。结论：滋养细胞表面FasL表达减少,引起母胎间的免疫耐受的破坏,是导致自然流产的一重量免疫病因,诱导FasL的产生或调节母胎间的免疫耐受将为临床治疗自然流产提供新的方向。     

孕激素对绒毛外滋养细胞VEGF表达的影响

目的通过研究孕激素对绒毛外滋养细胞孕激素受体(PR)和血管内皮生长因子表达的影响,探讨P对绒毛外滋养细胞侵袭能力的影响及维持妊娠的机理。方法采用半定量PCR和定量PCR测定孕激素干预后绒毛外滋养细胞表达PR mRNA和VEGF mRNA的水平。结果(1)不同浓度的孕激素PRA表达量极低,PRB mRNA的表达没有显著差异(P>0.05);(2)VEGF mRNA与孕激素浓度变化无明显相关,反而与PRB mRNA的水平成反比关系。结论(1)孕激素作用的发挥取决于PR的水平,与激素本身的浓度没有明显关系;(2)PRB能下调滋养细胞分泌VEGF的能力从而抑制其侵袭力,提示PR异常可能是导致绒毛外滋养细胞侵袭能力发生改变而引起相关疾病的原因之一。     

人早孕蜕膜组织及蜕膜基质细胞趋化因子受体及配体的转录特征

妊娠后，蜕膜化的子宫内膜（蜕膜，decidua）和发育的滋养细胞共同构成了母．胎界面。同种异体移植物的胎儿，能够在母体内存活直至足月分娩，这其中涉及母．胎界面极为复杂的免疫调节机制。蜕膜主要由蜕膜基质细胞（decidual stromal cell，DSC）（75％）和淋巴细胞构成，能分泌多种细胞因子，具有广泛的生物学功能，是母．胎界面来自母体的重要成分。     

母-胎界面趋化因子及其受体表达与作用

母-胎界面的滋养细胞、蜕膜淋巴细胞和蜕膜基质均表达多种趋化因子及其受体。滋养细胞分泌的MIP-1α募集外周血中CD56brightNK细胞至母-胎界面;蜕膜血管表达SLC,特异性趋化CD56brightNK细胞。除募集蜕膜淋巴细胞外,母-胎界面的趋化因子及其受体尚参与固有免疫应答,并调节滋养细胞侵袭、分化及胎盘形成。滋养细胞固有表达CXCR4和CCR5,HIV-1利用CXCR4或CCR5入侵胎儿细胞,导致HIV-1经子宫垂直传播。滋养细胞及蜕膜细胞分泌的IL-8在炎症相关的早产中具有重要作用。     

T细胞相关趋化因子及受体的研究进展

T细胞在发育、成熟、活化及发挥生物学效应的各个阶段表达不同的趋化因子受体。T细胞相关趋化因子及其受体的表达在不同的细胞类群上具有时相和分布的差异,并通过趋化因子与其受体特异性结合的模式,参与T细胞的发育过程,调控细胞的定向迁移,从而影响局部甚至整个机体的免疫状态。此外,它还在炎症、感染、肿瘤、自身免疫疾病等众多病理生理的过程中发挥重要作用。在这一领域的深入研究将为相关疾病的预防和治疗提供新的思路和途径。     

趋化因子在树突状细胞和T细胞迁移活化中的作用和意义

趋化因子是机体内一群能使细胞发生趋化运动的细胞因子，通过与表达于细胞表面的趋化因子受体结合而发挥其生物学作用。树突状细胞是一种专职的抗原递呈细胞。树突状细胞的成熟和功能与趋化因子和其受体介导的体内迁移密切相关，从而有效的将抗原递呈给初始T细胞并使之活化和分化，发挥其调节免疫应答的作用。     

早孕滋养细胞膜型及可溶性趋化因子CXCL16表达调控

目的 探讨趋化因子CXCL16在人早孕滋养细胞表达和释放的调控机制.方法 原代培养人早孕滋养细胞,实时定量RT-PCR、免疫化学和ELISA方法分析CXCL16的表达和分泌;ELISA方法分析细胞因子IFN-γ、TNF-α、IL-4刺激前后滋养细胞CXCL16的表达和分泌水平;ELISA方法分析ADAM10治疗前后滋养细胞CXCL16的表达及分泌水平.结果 滋养细胞表达并分泌趋化因子CXCL16;IFN-γ治疗后滋养细胞表达和分泌CXCL16水平均显著上升(P<0.01),但TNF-α和IL-4并不影响CXCL16的表达或分泌;ADAM10可以加速CXCL16自滋养细胞的脱落,但并不上调CXCL16的合成.结论 IFN-γ和ADAM10参与调控母胎界面滋养细胞趋化因子CXCL16的合成和分泌.     

子宫内膜及异位灶趋化因子受体转录在子宫内膜异位症发病中的作用

目的：探讨子宫内膜异位症患者在位内膜及异位灶18种趋化因子受体的转录特征，以揭示趋化因子受体及其配体在子宫内膜异位症发生发展中的作用。方法：以正常子宫内膜为对照，半定量RT-PCR检测子宫内膜异位症患者在位内膜及异位灶18种趋化因子受体mRNA的表达水平，并比较其差异。结果：与正常子宫内膜相比，子宫内膜异位症患者在位子宫内膜CCR6、CCR8、CCR9、CX3CR1表达明显升高（P＜0．05）。与在位内膜相比，异位灶CCR4、CCR8、CCR9、CXCR1表达显著升高（P＜0．05）。结论：在位子宫内膜CCR6、CCR8、CCR9、CX3CR1高表达，可能参与子宫内膜异位症的发生；异位灶CCR4、CCR8、CCR9、CXCR1高表达，可能参与子宫内膜异位症的进一步发展。     

人早孕母胎界面SOCS1、SOCS2、SOCS3表达

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 cytoldne 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、SOCS3 mRNA水平；Western blot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、IFN-γ。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。     

Expression of functional chemokine receptors of human placental cells

PROBLEM: Chemokine receptors of placental trophoblasts possibly act as co-receptors or alternative receptors of maternal fetal infection by HIV. To clarify their possible expression and the physiological roles of chemokines on human placentae, we studied chemokine chemokine receptor expression and the effects of exogenous chemokines on choriocarcinoma cell lines. MATERIALS AND METHODS: Placental samples were obtained from 13 placentae of various gestational ages. Villous tissue was mechanically dissected from samples. Trophoblasts were enriched by anti-human chorionic gonadotropin (hCG)-coated magnetic beads. Human choriocarcinoma cell lines (JAR, BeWo, JEG-3) were maintained in RPMI 1640 media supplemented with 10% FCS. Expression of chemokine receptors was studied by RT-PCR. The effects of MIP-1alpha, RANTES, MCP-1 on hCG production were estimated by EIA. Effects of chemokines on proliferation of choriocarcinoma cell lines were examined by MTT assay. RESULTS: We observed mRNA expression of CCR-1, 2, 3, 4, 5 and CXCR-1, 2, 4 in 1st trimester placental villi, CCR-I, 2, 4 and CXCR-1, 2. 4 in 2nd trimester placental villi, CCR-1, 2, 4 and CXCR-4 in 3rd trimester placental villi. Using MACS enriched trophoblasts, we observed identical results. A choriocarcinoma cell line BeWo expressed CCR-1, 3, 4 and CXCR-1, 2, 4 while JEG-3 and JAR expressed CCR-1, 3, 4, 5 and CXCR-1, 2, 4. Expression of the CCR-5 and CXCR-4 protein in choriocarcinoma cell lines and MACS-enriched trophoblats were confirmed by flow cytometry. Chemokine MCP-3, MIP-1alpha, RANTES mRNA were expressed by the 1st, 2nd and 3rd trimester placental samples and the three choriocarcinoma cell lines examined. MCP-1 was expressed by 1st and 2nd trimester placental villi. Administration of chemokines up-regulated proliferation (10(-1) - 10 ng/mL) and hCG production (10(-1) - 10(-2)ng/ mL) of the three choriocarcinoma cell lines examined. CONCLUSIONS: Our results suggest possible roles of chemokines/chemokine receptors on placental physiology and their involvement in HIV transmission as alternative receptors.     

Morphological interactions of human first trimester placental villi co-cultured with decidual explants

Abnormalities of pregnancy such as pre-eclampsia and intrauterinegrowth retardation are characterized by shallow trophoblasticinvasion of the placental bed, the precise molecular pathophysiologyof which remains to be fully elucidated. An in-vitro model involvinga co-culture of first trimester placental villi and deciduaparietalis explants (of 8-12 weeks gestation) was developedand used to characterize the migration and local invasion oftrophoblast cells. Trophoblast proliferation (confirmed by Ki-67immunostaining), differentiation and loose attachment of placentalvilli to the underlying decidual epithelium or stroma occurredwithin the first 24 h of co-culture. This was followed by erosionof the syncytial layer of the placental villi and commencementof a progressive cytotrophoblast invasion after 48 h of co-culture,which continued until 120 h, when the experiments were terminated.E-cadherin was expressed at the interfaces between trophoblastcells within the villi, but expression of this adhesion moleculeseemed to be down-regulated in the invasive trophoblast cells.Our results suggest that the model could be useful in investigatingthe factors that control early human placentation and the feto-maternalinterface.     

CXCR7分子对人早孕滋养细胞的调节作用

目的:已有研究表明CXCL12/CXCR4信号途径以自分泌的方式在人早孕滋养细胞增殖和信袭中发挥重要作用,探讨CXCL12的第二受体CXCR7信号对人早孕滋养细胞活力和侵袭功能的调节作用。方法:采用RT-PCR、免疫组化分析CXCR7在人早孕滋养细胞的表达;采用MTT和Transwell侵袭试验分别分析CXCL12/CXCR7信号途径对人早孕滋养细胞活力和侵袭功能的作用。结果:人早孕滋养细胞表达CXCR7分子,外源性给滋养细胞株HTR-8/Svneo添加anti-CXCR4(20μg/ml)和anti-CXCR7(10μg/ml)的中和性抗体后,均能显著抑制滋养细胞的增殖和侵袭能力。结论:人早孕滋养细胞表达CXCR7,通过CXCR7信号升调节滋养细胞的增殖和侵袭,其原因可能是与CXCR4形成二聚体共同参与CXCL12的信号传递。     

趋化因子及其受体与自身免疫性甲状腺疾病

趋化因子及其受体是免疫系统的重要组成部分,其趋化活性可以介导免疫细胞的定向移动和活化。在自身免疫性甲状腺疾病（AITD）的形成过程中,趋化因子及其受体参与淋巴细胞在甲状腺组织的浸润、定位及活化,它们的表达是导致AITD发生的一个重要因素。     

趋化因子及其受体在炎症中作用的研究进展

趋化因子是能够激发白细胞趋化性的小分子分泌性蛋白质,是可受化学诱导物及细胞因子调节、并能刺激细胞趋化运动的一类细胞因子.趋化因子将循环中的淋巴细胞募集到组织损伤或炎症反应部位.早期对趋化因子的研究是基于其在炎症中的功能.最近发现趋化因子及其受体和机体内单核细胞的迁移,适应性免疫应答以及多种疾病的发病机理相关.趋化因子受体作为炎症和免疫应答的调控分子而成为多种药物的靶向分子.临床上已经开始利用趋化因子的拈抗剂治疗炎症疾病,本篇旨在阐述趋化因子及其受体在炎症疾病中的作用以及这些趋化因子在临床上的应用.     

The fetal vascularisation of term human placental villi

Summary The fetal vascularisation of the most peripheral three generations of stem villi has been studied by means of semithin serial sectioning and scanning electron microscopy of vessel casts in 50 human term placentas. The vessel types have been classified according to the structure of the vessel walls. Stem villi with a diameter of 200–400 m are characterized by one smaller artery and one small vein, some arterioles and venules and capillaries of the paravascular net. Stem villi of about 150 m diameter contain arterioles and muscular venules besides the paravascular capillary network. The last generation of stem villi measuring 80 to 100 m in diameter exhibit a terminal arteriole and a collecting venule as well as up to ten paravascular capillaries. The luminal width of the arterial and venous stem vessels is considerably smaller than described for other vascular beds. This may partly be due to postpartal vascoconstriction. The capillaries of the paravascular net normally to not show sinusoidal dilation. They are arranged as long, hairpin-like, poorly branched loops connecting the arterial and venous stem vessels to each other.