“Real life” polycyclic aromatic hydrocarbon (PAH) mixtures modulate hCG,hPL and hPLGF levels and disrupt the physiological ratio of MMP-2 to MMP-9 and VEGF expression in human placenta cell lines

Using JEG-3 and BeWo cells, we examined the effect of “real life” mixtures of polycyclic aromatic hydrocarbons (PAHs), at doses reported in maternal blood (Mix I) and in placental tissue (Mix II), on human chorionic gonadotropin (hCG), placental lactogen (hPL) and placental growth factor (hPLGF) secretion, protein expression and immunolocalization. Additionally, the action of PAH mixtures on basal and hormone-stimulated matrix metalloproteinase-2 (MMP-2), MMP-9 and vascular endothelial growth factor (VEGF) protein expression was evaluated. Under basal conditions, the PAH mixtures increased hCG and decreased hPLGF levels in both cell lines, while hPL expression was stimulated in JEG-3 and inhibited in BeWo. There was no effect on the MMP-2/MMP-9 ratio or VEGF expression. In hormone-stimulated cells, PAH mixtures changed the MMP-2/MMP-9 ratio in JEG-3 cells in favor of MMP-9, while in BeWo MMP-2 was favored. The effect on VEGF expression was cell specific and dependent on the mixture. In hCG-treated cells, only Mix II inhibited VEGF expression in JEG-3 cells. Neither PAH mixtures affected this protein in BeWo cells. In hPL-treated cells, Mix I had a stimulatory effect in JEG-3 cells, while Mix II exerted an inhibitory effect in BeWo cells. In hPLGF-treated cells, Mix II decreased in JEG-3 cells, but in BeWo cells, both mixtures increased VEGF expression. Considering that the evaluated protein hormones play crucial roles in angiogenesis and neovascularization in the placenta, “real life” PAH mixtures by disrupting protein hormones levels, the MMP-2/MMP-9 ratio and VEGF expression can lead to insufficiency and many pregnancy-related disorders.     

17β-雌二醇对子宫内膜异位症患者在位子宫内膜间质细胞β-catenin mRNA和蛋白表达的影响

目的研究17β-雌二醇(17β-E2)对子宫内膜异位症(内异症)患者在位子宫内膜间质细胞β-catenin mRNA和蛋白表达的影响,探讨Wnt/β-catenin信号通路在介导雌激素促进内异症发生发展的作用。方法体外分离培养内异症患者在位子宫内膜间质细胞。用不同浓度17β-E2处理子宫内膜间质细胞48 h;此后选用10-10mol/L 17β-E2处理子宫内膜间质细胞12、24和48 h,逆转录聚合酶链反应(RT-PCR)和免疫印迹法(Western blotting)检测17β-E2处理前后子宫内膜间质细胞β-catenin mRNA和蛋白的表达水平。同法分析雌激素受体拮抗剂ICI182,780(10-6mol/L)对17β-E2促进β-catenin mRNA和蛋白表达的影响。免疫组织化学染色观察17β-E2作用后β-catenin在子宫内膜间质细胞中的定位。结果17β-E2能明显促进内异症患者在位子宫内膜间质细胞β-catenin mRNA和蛋白的表达,并呈剂量和时间依赖性,于10-10mol/L作用48 h最明显。雌激素受体拮抗剂ICI182,780能明显抑制17β-E2对子宫内膜间质细胞β-catenin mRNA和蛋白的表达。免疫组织化学染色发现17β-E2能促进β-catenin在子宫内膜间质细胞核内的表达。结论雌激素可能通过激活Wnt/β-catenin信号通路促进内异症在位子宫内膜的异位种植。     

人绒毛膜促性腺激素β亚基cDNA的克隆初步报告

用新鲜的人绒毛膜组织，抽提组织总ＲＮＡ并通过逆转录反应合成ｃＤＮＡ第一链。利用人工合成的一对寡核苷酸引物，采用ＰＣＲ技术特异性地扩增ｈＣＧ－β－ｃＤＮＡ。琼脂糖凝胶电泳结果显示扩增片段大小为５００ｂｐ，同预计的片段长度相符。将此片段利用Ｔ－Ａ载体克隆至ＰＣＲⅡ质粒上并进行全序列分析，结果显示克隆片段包含ｈＣＧ－βｃＤＮＡ５’端和３’端非编码区及完整的编码区。     

Plasminogen activator is involved in the hCG-induced neutrophil extravasation and vasopermeability increase in the rat testis

The role of proteolytic enzymes in the hCG-induced increase in testicular vasopermeability and neutrophil extravasation was studied using protease inhibitors. An intra-testicular injection of hCG together with incubation medium conditioned by polymorphonuclear leucocytes (PMNs) caused a significant increase in vasopermeability and a coincident extravasation of PMN's from the postcapillary venules in the rat testis. When p-aminobenzamidine, a serine protease inhibitor which inhibits urokinase-type plasminogen activator, was administered together with hCG in the incubation medium, both the permeability increase and PMN extravasation were prevented. Aprotinin, another serine protease inhibitor, and Eglin C, a specific neutrophil elastase and cathepsin G inhibitor were, however, without effect. None of these inhibitors caused any non-specific vascular effects in the testis at the concentrations used. These results support the concept that the hCG-induced increase in vasopermeability in the rat testis is related to extravasation of PMNs and suggest that urokinase-type plasminogen activator is involved in migration of these cells through the postcapillary venular walls.     

国产ELISA试剂盒应用于兴奋剂控制中的hCG检测

本文介绍用国产酶免定量药盒测定血、尿样中ｈＣＧ含量的一种ＥＬＩＳＡ方法。该方法特异性好、经济可靠而且简便快捷，适用于运动员尿样的兴奋剂检测。文中还报道了该方法应用于测定两例志愿者注射ｈＣＧ后的血、尿样及国际奥委会医学委员会１９９４年水平考试和１９９５年资格复试ｈＣＧ阳性尿样的结果，并讨论了测得值与其它试剂盒测定结果的差异及原因。     

hCG/ATP-induced Leydig cell calcium channel reaction at different extracellular calcium ion concentration

Objective: To study the calcium channel reaction of human Leydig cells induced by hCG/ATP at different extracellular calcium ion concentrations. Methods: The Leydig cell calcium ion concentration was examined with laser confocal microscope, when the cells were stimulated with hCG/ATP at different extracellular calcium contrations. Results: With calcium-containing extracellular fluid, the Leydig cells were sensitive to hCG stimulation and when the extracellular fluid was calcium-free, the Leydig cells did not respond to the stimulation. However, the Leydig cells did respond to ATP stimulation no matter the extracellular fluid contained calcium or not. Conclusion: In human Leydig cells, there are calcium channels sensitive to hCG and ATP. The extracellular calcium ion concentration plays an important role in the regulation of Leydig cell metabolism by hCG/ATP.     

大鼠睾丸间质细胞分离及其鉴定

目的和方法 :通过血管冲洗、Ⅱ型胶原酶消化以及Percoll梯度离心 ,建立大鼠睾丸间质细胞分离方法。结果 :(1)Percoll密度梯度离心后纯化的间质细胞 3β HSD染色阳性百分率可达 85 %。 (2 ) 1.2× 10 -3 IU/mlhCG可显著性增加间质细胞睾酮的分泌 (P <0 .0 5 )。结论 :提示该方法可靠且具有一定应用前景     

消炎痛对hCG诱发家兔排卵抑制的形态学研究

应用非甾体抗炎药消炎痛(20mg/kg)抑制hCG 诱发的家兔排卵,注射后的6、9、12、72小时剖腹观察排卵数,并取材进行透射电镜观察和酶组织化学观察。甾体激素合成酶类的组织化学和颗粒细胞内某些细胞器的电镜立体定量学测定结果表明:消炎痛抑制了排卵,却不影响颗粒细胞向黄体细胞的转变。透射电镜观察表明:在排卵前,实验组和对照组最明显的不同是:实验组的外膜和白膜很少见到胶原溶解现象,且表面上皮细胞内含的致密小体不释放其内容物。结果提示:消炎痛主要作用于排卵前卵泡顶部的成纤维细胞和表面上皮细胞,这可能与其抑制了某些与排卵有关的酶的释放有关。     

Melanoma cell adhesion molecule (MCAM/CD146) is expressed on human luteinizing granulosa cells: enhancement of its expression by hCG,interleukin-1 and tumour necrosis factor-alpha

Melanoma cell adhesion molecule (MCAM) was originally reported to be involved in the invasion and progression of melanoma. It was also shown to be responsible for the attachment of cells to endothelial cells. In this study, we demonstrated by immunohistochemistry that immunoreactive MCAM was not expressed on granulosa cells in the pre-ovulatory follicle, but it was clearly detected in large luteal cells in corpora lutea from the mid-luteal phase of the menstrual cycle. Northern blotting analysis confirmed the expression of MCAM mRNA in corpus luteum. MCAM was weakly detected by immunocytochemical staining in human luteinizing granulosa cells isolated from patients undergoing IVF treatment. Its expression was found to be increased during time in culture of these cells. Flow cytometry and Northern blot analysis revealed that MCAM expression on luteinizing granulosa cells was enhanced when the cells were cultured for 5 days in the presence of hCG (1 IU/ml) or cytokines such as interleukin-1alpha (10 ng/ml) and tumour necrosis factor-alpha (10 ng/ml). No significant difference of MCAM expression was observed between the cultures under normoxic (20% oxygen) and hypoxic (1% oxygen) conditions. These results indicate that luteinizing granulosa cells express MCAM and that MCAM expression is regulated by LH/hCG and cytokines during luteinization. Since MCAM has been reported to mediate cellular interaction with endothelial cells, this molecule may play a role in neovascularization during corpus luteum formation in the human ovary.     

TH1/TH2型细胞因子对人绒毛膜组织hCG分泌的影响

本研究应用人妊娠早期新鲜绒毛组织体外分泌人绒毛膜促性腺激素 (ｈＣＧ)的模型 ,探讨ＴＨ1 ＴＨ2 型细胞因子对ｈＣＧ分泌的影响 ,初步阐明ＴＨ1 型 ＴＨ2 型细胞因子在人类早期妊娠中的免疫调节作用 ,为今后临床诊断及治疗提供参考依据。1　材料与方法1 .1　主要试剂　ＲＰＭＩ 1 640 (美国ＧＩＢＣＯ公司 ) ,新生小牛血清 (ＮＣＳ) (医科院血研 ) ,人ＩＦＮγ、人ＩＬ 4(英国 ,ＰＥＰＲＯＴＥＣＨＥＣ有限公司 ) ,ｈＣＧ放射免疫分析药盒 (天津洁瑞公司 )。1 .2　绒毛组织悬液的制备　选择经临床确诊为正常妊娠的6～ 1 2周龄的早孕妇女…