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Vaginal matrix metalloproteinase levels in pregnant women with bacterial vaginosis. 细菌性阴道病孕妇阴道基质金属蛋白酶水平。
Pub Date : 2006-09-01 Epub Date: 2006-07-26 DOI: 10.1016/j.jsgi.2006.05.008
Laura Diaz-Cueto, Adrian Cuica-Flores, Francisco Ziga-Cordero, Jose Antonio Ayala-Mendez, Gilberto Tena-Alavez, Pablo Dominguez-Lopez, Rocio Cuevas-Antonio, Fabian Arechavaleta-Velasco

Objective: To compare matrix metalloproteinase (MMP)-8 and MMP-9 levels in the vaginal secretions of pregnant women with or without asymptomatic bacterial vaginosis (BV).

Methods: In this study, vaginal levels and molecular forms of MMP-8 and MMP-9 were studied in 36 pregnant women between 28 and 34 weeks of gestation with asymptomatic BV and 41 pregnant women, matched for gestational age, without BV.

Results: Vaginal MMP-8 concentrations were significantly higher (P = .023) in BV-positive women. There was no significant difference in MMP-9 levels between healthy pregnant controls and BV-positive pregnant women. The presence of MMP-8 was confirmed by a 38-kd band on Western blots.

Conclusions: Our findings show that BV is associated with increased levels of MMP-8 in vaginal fluid. Increased production of collagen-degrading enzymes such as MMP-8 is a possible cause of spontaneous preterm delivery in pregnant women with asymptomatic BV.

目的:比较有无无症状细菌性阴道病(BV)孕妇阴道分泌物中基质金属蛋白酶(MMP)-8和MMP-9的水平。方法:本研究对36例妊娠28 ~ 34周无症状BV孕妇和41例胎龄匹配的无BV孕妇阴道中MMP-8和MMP-9的水平和分子形态进行了研究。结果:bv阳性妇女阴道MMP-8浓度显著增高(P = 0.023)。bv阳性孕妇与健康孕妇间MMP-9水平无显著差异。Western blots上38-kd条带证实了MMP-8的存在。结论:我们的研究结果表明细菌性阴道炎与阴道液中MMP-8水平升高有关。胶原降解酶如MMP-8的产生增加是无症状BV孕妇自发性早产的可能原因。
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引用次数: 25
Endothelial activation and cell adhesion molecule concentrations in pregnant women living at high altitude. 高海拔地区孕妇内皮细胞活化及细胞黏附分子浓度。
Pub Date : 2006-09-01 Epub Date: 2006-07-31 DOI: 10.1016/j.jsgi.2006.05.001
Leah Marks, Stacy Zamudio, Frances Cousins, Elizabeth Duffie, Fiona Lyall

Objectives: Maternal physiology at high altitude could be considered to resemble an intermediate state between preeclampsia and normal pregnancy. The objective of the current study was to determine if cell adhesion molecules, known to be increased in preeclampsia, are increased with chronic maternal and placental hypoxia (due to high-altitude residence) in the absence of preeclampsia.

Methods: Serum was collected from women residing at 3100 m or 1600 m in the three trimesters of pregnancy and postpartum. Vascular cell adhesion molecule-1 (VCAM-1), E-selectin, and intercellular adhesion molecule-1 (ICAM-1) were measured by enzyme-linked immunosorbent assay (ELISA).

Results: General linear model (GLM) repeated measures analysis of VCAM-1, E-selectin, and ICAM-1 data showed there were no statistically significant effects of gestation within either the high- or moderate-altitude groups or between the different altitudes.

Conclusion: The increase in cell adhesion molecules reported in preeclampsia is not present in pregnant women at high altitude, suggesting that maternal systemic hypoxia is not responsible for this pathway of endothelial cell activation in preeclampsia.

目的:高海拔地区产妇的生理状态类似于子痫前期和正常妊娠之间的一种中间状态。当前研究的目的是确定在没有子痫前期的情况下,已知在子痫前期增加的细胞粘附分子是否在母体和胎盘慢性缺氧(由于高海拔居住)时增加。方法:在妊娠3个月和产后居住在3100米和1600米的妇女中采集血清。采用酶联免疫吸附试验(ELISA)检测血管细胞粘附分子-1 (VCAM-1)、e-选择素和细胞间粘附分子-1 (ICAM-1)。结果:VCAM-1、E-selectin和ICAM-1数据的一般线性模型(GLM)重复测量分析显示,高、中海拔组和不同海拔组之间妊娠无统计学意义。结论:高海拔孕妇在子痫前期报告的细胞粘附分子的增加不存在,提示母体全身性缺氧不是子痫前期内皮细胞激活这一途径的原因。
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引用次数: 8
Up-regulation of the endothelin receptor A in placental tissue from first trimester delayed miscarriages. 妊娠早期迟发性流产胎盘组织内皮素受体A的上调。
Pub Date : 2006-09-01 Epub Date: 2006-07-31 DOI: 10.1016/j.jsgi.2006.05.006
Martina Dieber-Rotheneder, Christina Stern, Gernot Desoye, Mila Cervar-Zivkovic

Objective: This study tested the hypothesis that the endothelin (ET)/ET receptor (ETR) system in biologic fluids and in the human placenta is altered in delayed miscarriages as compared to apparently normal early pregnancies (reference group).

Methods: Immunoreactive ET (irET) concentrations were measured in plasma, urine, and cervical smears from 57 pregnant women in the weeks 6 to 14 of gestation (46 delayed miscarriages, 11 references) with radioimmunoassay (RIA). ET-1, ETR-A, and ETR-B mRNA, and ETR protein expression were measured in placental tissue of 45 early pregnancies (31 delayed miscarriages, 14 references) using semiquantitative reverse-transcription polymerase chain reaction (RT-PCR) and immunoblotting, respectively.

Results: irET levels in plasma, urine, and cervical smears did not differ between groups. Two prevailing ETR-A and ETR-B proteins were found at 45 and 55 kd, and were distributed similarly in delayed miscarriages and references. ETR-A protein and mRNA levels were 54% (P = .009) and threefold (P = .021) higher, respectively, in delayed miscarriages versus references. There was no difference in placental ETR-B and ET-1 mRNA levels between groups.

Conclusion: Neither irET nor ET-1 mRNA levels differ between delayed miscarriages and normal early pregnancies. Pregnancies at risk for miscarriage cannot be identified by measurement of ET in plasma, urine, or cervical smears. Within the ET/ETR system, ETR-A is selectively up-regulated in placental tissue of delayed miscarriages as compared to normal pregnancies. ETR protein processing is similar in both groups.

目的:本研究验证了生物体液和人胎盘中的内皮素(ET)/ET受体(ETR)系统在延迟流产中与明显正常的早孕(参照组)相比发生改变的假设。方法:应用放射免疫分析法(RIA)测定57例妊娠6 ~ 14周(46例迟发性流产,11例文献)孕妇的血浆、尿液和子宫颈涂片中的免疫反应性ET (irET)浓度。采用半定量反转录聚合酶链反应(RT-PCR)和免疫印迹法分别检测45例早孕(31例迟发性流产,14篇文献)胎盘组织中ET-1、et - a、et - b mRNA和ETR蛋白的表达。结果:血浆、尿液和子宫颈涂片中的irET水平在两组之间没有差异。在45和55 kd处发现了两种主要的etra和etrb蛋白,并且在延迟流产和参考文献中分布相似。延迟流产组的etra蛋白和mRNA水平分别比对照组高54% (P = 0.009)和3倍(P = 0.021)。各组间胎盘et - b和ET-1 mRNA水平无差异。结论:irET和ET-1 mRNA水平在延迟性流产和正常早孕中均无差异。有流产风险的妊娠不能通过测定血浆、尿液或子宫颈涂片中的ET来确定。在ET/ETR系统中,与正常妊娠相比,延迟流产的胎盘组织中ET - a选择性上调。两组的ETR蛋白加工过程相似。
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引用次数: 6
The Future of the Journal of the Society for Gynecologic Investigation 妇科调查学会杂志的未来
Pub Date : 2006-09-01 DOI: 10.1016/j.jsgi.2006.07.007
H. Taylor
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引用次数: 0
Systemic activation and vascular infiltration of neutrophils with term labor. 足月分娩时中性粒细胞的全身活化和血管浸润。
Pub Date : 2006-09-01 DOI: 10.1016/j.jsgi.2006.06.001
Courtney E Leik, Scott W Walsh

Objective: The purpose of the current study was to determine if neutrophils infiltrate maternal systemic vascular tissue at the time of term labor.

Methods: Subcutaneous fat biopsies were obtained at cesarean delivery or abdominal surgery from laboring women (n = 5), non-laboring women (n = 5), and normal non-pregnant women (n = 5). Immunohistochemical staining was performed for CD66b, a neutrophil antigen, and intercellular adhesion molecule-1 (ICAM-1; CD54), an endothelial cell adhesion molecule for neutrophils. Vessels (10 to 200 microm) were analyzed for intensity of staining and percentage of vessels with staining.

Results: CD66b staining intensity was significantly greater for laboring women at term than for non-laboring women at term or for normal non-pregnant women (1.3 +/- 0.3 versus 0.2 +/- 0.1 versus 0.2 +/- 0.1, respectively, P < .01). Laboring women had significantly more vessels with staining for CD66b (79 +/- 4 versus 24 +/- 8 versus 19 +/- 6%, P < .001), more vessels with neutrophils adhered and flattened to endothelium (67 +/- 3 versus 16 +/- 7 versus 12 +/- 4%, P < .001), more vessels with neutrophils in the intima (30 +/- 6 versus 5 +/- 2 versus 2 +/- 1%, P < .05), and a greater number of neutrophils per vessel (5.4 +/- 1.1 versus 1.7 +/- 0.5 versus 1.2 +/- 0.3, P < .01) as compared to non-laboring or normal non-pregnant women. ICAM-1 staining was present in the endothelium of all groups, with no difference in staining intensity or percent of vessels stained. Between 86% to 96% of vessels stained for ICAM-1. Laboring patients had numerous leukocytes stained for ICAM-1 in their vessels.

Conclusion: This study demonstrates that neutrophils infiltrate maternal systemic vascular tissue at the time of term labor. Neutrophils were flattened and adhered to endothelium and infiltrated into the intimal space.

目的:本研究的目的是确定中性粒细胞是否在足月分娩时渗入母体全身血管组织。方法:在剖宫产或腹部手术中分别对产妇(n = 5)、非产妇(n = 5)和正常非孕妇(n = 5)进行皮下脂肪活检。对CD66b、中性粒细胞抗原和细胞间粘附分子-1 (ICAM-1)进行免疫组化染色;CD54),中性粒细胞的内皮细胞粘附分子。分析血管(10 ~ 200微米)染色强度和染色血管百分比。结果:产程妇女足月CD66b染色强度显著高于非产程妇女足月或正常非妊娠妇女(分别为1.3 +/- 0.3 vs . 0.2 +/- 0.1 vs . 0.2 +/- 0.1, P < 0.01)。劳动妇女有更显著的血管与染色CD66b(79 + / - 4和24 + / - 8和19 + / - 6%,P <措施),更多的船只与中性粒细胞粘附和扁平的内皮(67 + / - 3和16 + / - 7和12 + / - 4%,P <措施),血管内膜的中性粒细胞(30 + / - 6和5 + / - 2和2 + / - 1%,P < . 05),每船大量中性粒细胞(5.4 + / - 1.1和1.7 + / - 0.5和1.2 + / - 0.3,P < 0.01)。各组内皮细胞均有ICAM-1染色,染色强度和血管染色百分比无差异。86% - 96%的血管有ICAM-1染色。分娩患者的血管中有大量白细胞染色显示ICAM-1。结论:本研究表明,中性粒细胞在足月分娩时浸润母体全身血管组织。中性粒细胞变平,粘附在内皮细胞上,向内膜浸润。
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引用次数: 5
Female infertility and free radicals: potential role in adhesions and endometriosis. 女性不孕症和自由基:粘连和子宫内膜异位症的潜在作用。
Pub Date : 2006-09-01 Epub Date: 2006-07-26 DOI: 10.1016/j.jsgi.2006.05.002
Zeynep Alpay, Ghassan M Saed, Michael P Diamond

Free radicals are highly reactive molecules produced in the cell either as part of, or as end-products of, biochemical reactions that have crucial roles in the homeostasis of the organism. Thus, excess production or impaired elimination of free radicals leads to increased oxidative stress, which has been implicated in the development of several different disease states, including hypoxia-reperfusion injury, cancer, and aging. Peritoneal adhesions and endometriosis are relatively commonly identified in women, and are known to be associated with infertility without clearly understood pathophysiology. The prevention and treatment strategies of these conditions, both of which have tremendous propensity to recur, have not been completely established. The development of both disorders has been shown to be closely related to the presence of increased oxidative stress in the tissues. In this article, we review this relationship with reference to the mechanistic steps involved and their regulation. As our knowledge of both conditions expands, we believe there will be opportunities for specific steps to intervention in free radical metabolism to reduce and/or prevent further development of endometriosis and adhesions.

自由基是细胞内产生的高活性分子,是生物化学反应的一部分或最终产物,在生物体的体内平衡中起着至关重要的作用。因此,自由基的过量产生或消除受损导致氧化应激增加,这与几种不同疾病状态的发展有关,包括缺氧再灌注损伤、癌症和衰老。腹膜粘连和子宫内膜异位症在女性中比较常见,并且已知与不孕症有关,但没有明确的病理生理学。这些疾病的预防和治疗策略都没有完全确定,这两种疾病都有很大的复发倾向。这两种疾病的发展已被证明与组织中氧化应激增加的存在密切相关。在这篇文章中,我们回顾了这种关系,涉及的机械步骤和他们的调节。随着我们对这两种情况的了解的扩大,我们相信将有机会采取具体措施干预自由基代谢,以减少和/或防止子宫内膜异位症和粘连的进一步发展。
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引用次数: 129
Prolonged mild hypoxia alters fetal sheep electrocorticogram activity. 长时间轻度缺氧会改变胎羊的皮质电图活动。
Pub Date : 2006-09-01 Epub Date: 2006-07-31 DOI: 10.1016/j.jsgi.2006.05.007
Victor M Pulgar, Jie Zhang, G Angela Massmann, Jorge P Figueroa

Objective: To assess the effects of prolonged mild hypoxemia on fetal brain electrocorticogram (ECoG) in late gestation.

Study design: Fetal and maternal catheters were placed under general anesthesia and animals allocated at random to receive intratracheal maternal administration of either nitrogen (n = 8) or compressed air (n = 8). Five days after surgery (125 days' gestational age), nitrogen infusion was adjusted to reduce fetal brachial artery PO2 by 25%. The targeted decrease in fetal oxygenation was maintained for 5 days while fetal ECoG activity and fetal and maternal cardiovascular variables were continuously recorded. Data are presented as mean +/- SEM and were analyzed by two-way analysis of variance (ANOVA) or two-sample t test.

Results: Nitrogen infusion decreased fetal Po2 by 26% (20.5 +/- 1.7 versus 14.3 +/- 0.8) without changing fetal PCO2 or pH. Mild fetal hypoxemia was associated with fetal tachycardia and increased fetal blood pressure (P < .05). Fetal ECoG in hypoxic fetuses showed a significant decrease in the time spent in high voltage (HV) (P < .05) and an increase in the time spent in low voltage (LV) and in the number of low voltage events (P < .05). Also, a significant decrease in the proportion of 1-4 Hz and an increase in the proportion of 13-20 Hz frequencies was observed in LV events without a significant change in the frequency profile of HV events (P < .05).

Conclusion: Prolonged mild hypoxemia significantly altered fetal homeostasis as reflected by the sustained tachycardia and increased blood pressure. Fetal ECoG activity was affected significantly in a qualitatively and quantitative manner by mild prolonged hypoxemia.

目的:探讨妊娠后期长期轻度低氧血症对胎儿脑皮质电图(ECoG)的影响。研究设计:全身麻醉下放置胎儿和母体导管,随机分配动物接受母体气管内输注氮气(n = 8)或压缩空气(n = 8)。手术后5天(孕龄125天),调整输注氮气使胎儿肱动脉PO2降低25%。在持续记录胎儿ECoG活动和胎儿及母体心血管变量的同时,维持胎儿氧合水平的定向降低5天。数据以均数+/- SEM表示,并通过双向方差分析(ANOVA)或双样本t检验进行分析。结果:输注氮可使胎儿Po2降低26% (20.5 +/- 1.7 vs 14.3 +/- 0.8),而不改变胎儿PCO2或ph。轻度胎儿低氧血症与胎儿心动过速和胎儿血压升高相关(P < 0.05)。低氧胎儿的ECoG显示高电压(HV)持续时间显著减少(P < 0.05),低电压(LV)持续时间和低电压事件次数显著增加(P < 0.05)。此外,在LV事件中观察到1-4 Hz频率的比例显著降低,13-20 Hz频率的比例显著增加,而HV事件的频率分布没有显著变化(P < 0.05)。结论:长期轻度低氧血症显著改变胎儿体内平衡,表现为持续的心动过速和血压升高。轻度长时间低氧血症对胎儿脑电图活动有显著的定性和定量影响。
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引用次数: 9
Hyperpolarization-activated cation channels are expressed in rat hypothalamic gonadotropin-releasing hormone (GnRH) neurons and immortalized GnRH neurons. 超极化激活的阳离子通道在大鼠下丘脑促性腺激素释放激素(GnRH)神经元和永生化GnRH神经元中表达。
Pub Date : 2006-09-01 Epub Date: 2006-07-31 DOI: 10.1016/j.jsgi.2006.05.010
Armando Arroyo, Beomsu Kim, Randall L Rasmusson, Glenna Bett, John Yeh

Objectives: The current research was conducted to determine whether hyperpolarization-activated cyclic nucleotide-gated (HCN1-4) channels are expressed in gonadotropin-releasing hormone (GnRH) neurons in the female rat hypothalamus and immortalized GnRH neurons (GT1-7 cells).

Methods: Double-label fluorescence immunohistochemistry was used to colocalize HCN1-4 channels and GnRH in GnRH neurons in the female rat hypothalamus. Reverse transcriptase-polymerase chain reaction (RT-PCR), Western blotting, and immunocytochemistry were used to analyze HCN channel gene expression in GT1-7 cells.

Results: Double-label fluorescence immunohistochemistry showed that 43% of hypothalamic GnRH neurons immunostained for HCN2 and 90% of GnRH neurons immunostained for HCN3. RT-PCR and Western blot showed expression of all four HCN channel subunits in GT1-7 cells. Double-label immunocytochemistry showed cytoplasmic immunostaining of HCN2 and HCN3 in GT1-7 cells.

Conclusions: This study demonstrates for the first time that HCN channels are expressed in GnRH neurons in the rat hypothalamus and GT1-7 cells. Our research supports the hypothesis that HCN channels may be involved in electrical bursting activity and pulsatile GnRH secretion in endogenous GnRH neurons and GT1-7 cells.

目的:本研究旨在确定超极化激活的环核苷酸门控(HCN1-4)通道是否在雌性大鼠下丘脑促性腺激素释放激素(GnRH)神经元和永生化GnRH神经元(GT1-7细胞)中表达。方法:采用双标记荧光免疫组织化学方法对雌性大鼠下丘脑GnRH神经元中的HCN1-4通道和GnRH进行共定位。采用逆转录聚合酶链反应(RT-PCR)、Western blotting和免疫细胞化学分析GT1-7细胞中HCN通道基因的表达。结果:双标记荧光免疫组化显示下丘脑GnRH神经元HCN2免疫染色43%,HCN3免疫染色90%。RT-PCR和Western blot均显示GT1-7细胞中4种HCN通道亚基均有表达。双标记免疫细胞化学显示GT1-7细胞中HCN2和HCN3的胞浆免疫染色。结论:本研究首次证实了HCN通道在大鼠下丘脑和GT1-7细胞的GnRH神经元中表达。我们的研究支持了HCN通道可能参与内源性GnRH神经元和GT1-7细胞的电爆发活动和脉冲性GnRH分泌的假设。
{"title":"Hyperpolarization-activated cation channels are expressed in rat hypothalamic gonadotropin-releasing hormone (GnRH) neurons and immortalized GnRH neurons.","authors":"Armando Arroyo,&nbsp;Beomsu Kim,&nbsp;Randall L Rasmusson,&nbsp;Glenna Bett,&nbsp;John Yeh","doi":"10.1016/j.jsgi.2006.05.010","DOIUrl":"https://doi.org/10.1016/j.jsgi.2006.05.010","url":null,"abstract":"<p><strong>Objectives: </strong>The current research was conducted to determine whether hyperpolarization-activated cyclic nucleotide-gated (HCN1-4) channels are expressed in gonadotropin-releasing hormone (GnRH) neurons in the female rat hypothalamus and immortalized GnRH neurons (GT1-7 cells).</p><p><strong>Methods: </strong>Double-label fluorescence immunohistochemistry was used to colocalize HCN1-4 channels and GnRH in GnRH neurons in the female rat hypothalamus. Reverse transcriptase-polymerase chain reaction (RT-PCR), Western blotting, and immunocytochemistry were used to analyze HCN channel gene expression in GT1-7 cells.</p><p><strong>Results: </strong>Double-label fluorescence immunohistochemistry showed that 43% of hypothalamic GnRH neurons immunostained for HCN2 and 90% of GnRH neurons immunostained for HCN3. RT-PCR and Western blot showed expression of all four HCN channel subunits in GT1-7 cells. Double-label immunocytochemistry showed cytoplasmic immunostaining of HCN2 and HCN3 in GT1-7 cells.</p><p><strong>Conclusions: </strong>This study demonstrates for the first time that HCN channels are expressed in GnRH neurons in the rat hypothalamus and GT1-7 cells. Our research supports the hypothesis that HCN channels may be involved in electrical bursting activity and pulsatile GnRH secretion in endogenous GnRH neurons and GT1-7 cells.</p>","PeriodicalId":17373,"journal":{"name":"Journal of the Society for Gynecologic Investigation","volume":"13 6","pages":"442-50"},"PeriodicalIF":0.0,"publicationDate":"2006-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/j.jsgi.2006.05.010","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"26176187","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 22
Redox-induced apoptosis of human oocytes in resting follicles in vitro. 氧化还原酶诱导的体外静息卵泡卵母细胞凋亡。
Pub Date : 2006-09-01 Epub Date: 2006-07-31 DOI: 10.1016/j.jsgi.2006.05.005
Xiao Zhang, Xiao-Hong Li, Xu Ma, Zhao-Huei Wang, Shuang Lu, Ying-Lu Guo

Background: The age-related depletion of the resting follicle (RF) stock occurs as a result of two processes: atresia and entry in growth phase. Due to difficulties in obtaining sufficient numbers of RFs for study, little is known about the apoptotic mechanisms for RF atresia. The present study was designed to investigate the effects of oxidative stress on the apoptosis of RF oocytes.

Methods: RFs isolated from human adult ovaries were cultured in vitro, treated with H2O2 at various concentrations (50 microM, 100 microM, 1.0 mM) for 1 hour, and observed for up to 48 hours. The oxidant-induced apoptosis of oocytes were observed by detection of DNA fragments, mitochondria membrane potential (MMP), and cytochrome c release.

Results: Based on nuclear morphology and TUNEL (terminal deoxynucleotidyl transferase-mediated dDTP nick end-labeling), oocyte apoptosis was observed in the RFs treated with 50 microM H2O2 with rates of 35% and 43% at 24 and 48 h after treatment, respectively. But intensive oxidative stress (1 mM H2O2) caused mainly necrosis as measured by quantifying propidium iodide (PI)-positive oocytes (44% within 12 hours), with lower level of apoptosis (17%) being observed at 24 hours after treatment. RFs treated with 100 microM H2O2 showed both apoptosis with the similar rate observed at 50microM and necrosis (13% PI-positive oocytes). Although pre-incubation with cyclosporine A (CsA) could effectively prevent oxidant-induced MMP collapse, but failed to suppress apoptosis of oocytes in RFs.

Conclusions: Oocytes of RFs in adult ovaries retain their ability to undergo apoptosis under oxidative stress, which is both dose- and time-dependent.

背景:与年龄相关的静息卵泡(RF)存量耗竭是两个过程的结果:闭锁和进入生长期。由于难以获得足够数量的RF用于研究,因此对RF闭锁的凋亡机制知之甚少。本研究旨在探讨氧化应激对RF卵母细胞凋亡的影响。方法:体外培养成人卵巢RFs,用不同浓度H2O2(50微米、100微米、1.0毫米)处理1小时,观察48小时。通过检测DNA片段、线粒体膜电位(MMP)和细胞色素c的释放,观察氧化诱导的卵母细胞凋亡。结果:基于细胞核形态学和TUNEL(末端脱氧核苷酸转移酶介导的dDTP缺口末端标记),50 μ m H2O2处理后24和48 h,卵母细胞凋亡率分别为35%和43%。但通过对碘化丙啶(PI)阳性卵母细胞的定量测量,强化氧化应激(1 mM H2O2)主要导致坏死(12小时内占44%),在处理后24小时观察到较低水平的凋亡(17%)。RFs经100 μ m H2O2处理后,细胞凋亡和坏死(pi阳性卵母细胞占13%)的比例与50 μ m处理时相似。环孢素A (cyclosporine A, CsA)预孵育虽能有效预防氧化诱导的MMP塌陷,但不能抑制RFs卵母细胞的凋亡。结论:在氧化应激条件下,成年卵巢中RFs的卵母细胞仍具有细胞凋亡的能力,且细胞凋亡具有剂量依赖性和时间依赖性。
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引用次数: 83
Progestagenic effects of tibolone are target gene-specific in human endometrial cells. 替博龙对人子宫内膜细胞的孕激素作用是靶基因特异性的。
Pub Date : 2006-09-01 Epub Date: 2006-07-31 DOI: 10.1016/j.jsgi.2006.05.004
Wei Zhang, James Mazella, Helenius J Kloosterboer, Linda Tseng

Objectives: Tibolone (Tib) exhibits progestagenic activities in addition to its tissue-specific estrogenic activities. The purpose of the current study was to determine the progestagenic actions of Tib and its metabolites using target genes known to be regulated by progestins in human endometrial glandular and stromal cells.

Methods: Human endometrial glandular and stromal cells were isolated from endometrial tissue fragments and separately incubated with Tib and its metabolites. Real-time polymerase chain reaction (PCR) was used to determine the mRNA content of 17betahydroxy steroid dehydrogenase (17betaHSD, type 2) and sulfotransferase (SULT1E1) in endometrial glandular cells, and prolactin (PRL) and insulin-like growth factor binding protein-1 (IGFBP1) in endometrial stromal cells.

Results: In glandular cells, Tib and Delta4-tibolone (Delta4Tib) significantly increased the content of 17betaHSD and SULT1E1 mRNA. In stromal cells, Tib and Delta4Tib increased PRL mRNA ( approximately 30% of the capacity compared to progesterone) and had little effect on IGFBP1 mRNA. Anti-progestin, RU486, reversed the induction of SULT1E1 and PRL by progesterone or Tib. Also, the two 3 hydroxyl tobolone metabolites, especially 3betaOHTib, showed some progestagenic effects.

Conclusions: The data showed that Tib and Delta4Tib exhibited clear progestagenic effects in endometrial glandular cells by inducing 17betaHSD and SULT1E1, while in stromal cells the response was weaker in the induction of PRL and had little effect on IGFBP1. In addition, the 3betaOHTib metabolite expressed progestagenic activity. These disparate effects in two types of cells may be beneficial for maintaining endometrial cells in a quiescent state.

目的:Tibolone (Tib)除了具有组织特异性雌激素活性外,还具有孕激素活性。本研究的目的是利用已知在人子宫内膜腺细胞和基质细胞中受孕激素调节的靶基因来确定Tib及其代谢物的孕激素作用。方法:从子宫内膜组织碎片中分离人子宫内膜腺细胞和基质细胞,分别与Tib及其代谢物孵育。采用实时聚合酶链反应(Real-time polymerase chain reaction, PCR)检测子宫内膜腺细胞中17betaHSD脱氢酶(17betaHSD, type 2)和硫转移酶(SULT1E1) mRNA含量,以及子宫内膜基质细胞中泌乳素(PRL)和胰岛素样生长因子结合蛋白-1 (IGFBP1) mRNA含量。结果:在腺细胞中,Tib和Delta4-tibolone (Delta4Tib)显著增加了17betaHSD和SULT1E1 mRNA的含量。在基质细胞中,Tib和Delta4Tib增加了PRL mRNA(与黄体酮相比,其容量约为30%),而对IGFBP1 mRNA的影响很小。抗黄体酮RU486逆转了黄体酮或Tib对SULT1E1和PRL的诱导。另外,两种3羟基羟溴酮代谢物,特别是3betaOHTib,也显示出一定的孕激素作用。结论:数据显示,Tib和Delta4Tib通过诱导17betaHSD和SULT1E1对子宫内膜腺细胞有明显的孕激素作用,而在基质细胞中,对PRL的诱导作用较弱,对IGFBP1的影响较小。此外,3betaOHTib代谢物表达孕激素活性。这两种细胞的不同作用可能有利于维持子宫内膜细胞处于静止状态。
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引用次数: 4
期刊
Journal of the Society for Gynecologic Investigation
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