反义寡脱氧核苷酸及卵泡刺激素对卵巢黏液性囊腺癌细胞增殖细胞核抗原表达的影响

目的 观察卵泡刺激素受体（FSHR）反义寡脱氧核苷酸（antisense ODN）及卵泡刺激素（FSH）对体外培养人卵巢黏液性囊腺癌（OMC）细胞增殖细胞核抗原（PCNA）表达的影响。方法 OMC细胞与不同浓度的FSH、antisense ODN和无义ODN（onnsensc ODN）共培育48、72h后．采用免疫细胞化学SP法检测细胞核内PCNA的表达。结果FSH明显增强PCNA的表达，而antiscnse ODN则显著降低PCNA的表达．与对照组比较．差异均有显著性意义（均P＜0．05或P＜0．01）；应用nonsensc,ODN未观察到其对PCNA表达的影响。同时．antisense ODN可明显拮抗FSH促PCNA表达增加的作用，差异有极显著性意义（P＜0．01）。结论 在OMC发展过程中，FSH具有一定的促进作用．antisense ODN可有效抑制癌细胞的增殖以及FSH的促增殖作用。     

Antisense oligodeoxynucleotide inhibits expression of recombinant porcine follicle-stimulating hormone receptor

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Effects of Progestin and Antiprogestin on the Expression of FSH Receptor and LH Receptor mRNA in Porcine Granulosa and Thecal Cells

We havereported thattheprogestin and antiprogestin both inhibited secretion ofsteroid by porcine granulosa and thecal cells cultured in a dual chambersystem.Theprevious results suggested that this inhibition seemed to be via reduction of go-nadotropin stimulation[1 ,2 ] . The anterior pituitary cell types are responsible for thesynthesis and secretion of LH and FSH known as gonadotropes. LH and FSH exerttheir effects through activation of gonadotropin receptors present on the plasmamembran…     

Effects of Progestin and Antiprogestin on the Expression of FSH Receptor and LH Receptor mRNA in Porcine Granulosa and Thecal Cells

In order to investigate the mechanism of progestin and antiprogestin in the regula tion of ovarian steroidogenesis, a dual-chamber culture system was prepared with the amnion membrane of human placenta. Isolated porcine granulosa and thecal cells from 4～6 mm-diameter follicles were grown on both sides of the amnion, respectively, and co-cultured with or without LNG and RU486. After 48 h incubation, the mRNAs of FSH receptor (FSH-R) and LH receptor (LH-R) of both cells were observed by in situ hybridization. The results showed that granulosa cells expressed both FSH-R mR NA and LH-R mRNA, while thecal cells expressed LH-R mRNA only. Under the stimulation of FSH, both LNG and RU486 increased FSH-R mRNA expression of granulosa cells. Under the stimulation of LH, LNG enhanced LH-R mRNA expres sion of thecal cells; while RU486 decreased its expression. When granulosa and thecal cells were exposed to FSH and LH both, the actions of LNG and RU486 in thecal cells showed the same result as that stimulated by LH alone. In granulosa cells LNG de creased LH-R mRNA expression, while RU486 increased its expression. These data suggest that: (1) granulosa cells expressed FSH-R mRNA significantly;(2) both the progestin and antiprogestin directly acted on the mRNA expression of gonadotropin re ceptors of ovarian cells, but effects were different; (3) the response of granulosa or thecal cells to the action of LNG and RU 486 was not the same. The mechanism needs to be further investigated.     

来曲唑诱导多囊卵巢综合征大鼠卵巢组织中FSH受体的表达及其意义

目的：研究来曲唑诱导的多囊卵巢综合征（PCOS）模型大鼠卵巢组织中促卵泡刺激素受体(FSHR)基因的表达,为该模型的应用提供一定的理论依据。方法：40只雌性SD大鼠随机分为模型组和对照组,每组20只,模型组应用来曲唑诱导大鼠PCOS模型,对照组不作任何处理。应用放射免疫法测定大鼠血清性激素水平;HE染色观察卵巢组织学变化,Real-time-PCR、免疫组织化学和Western blotting法检测FSHR基因在卵巢组织中的表达。结果：与对照组比较,模型组血清睾酮(T)、黄体生成素(LH)、卵泡刺激素(FSH)浓度显著增高（P<0.05）,雌二醇(E2)、孕酮(P)浓度降低（P<0.05）。卵巢质量两组间比较差异无显著性(P>0.05)。与对照组比较,模型组可见囊状扩张卵泡明显增多,卵巢白膜增厚,黄体数量明显减少。模型组卵巢组织中FSHR mRNA及蛋白表达水平均显著高于对照组（P<0.05）。结论：来曲唑诱导的PCOS大鼠卵巢组织中FSHR基因表达水平与人类PCOS患者相似,该模型是研究PCOS病理机制的一种理想的动物模型。     

Further evidence for differential regulation of follicle-stimulating hormone (FSH) and luteinizing hormone (LH): increased FSH and decreased LH levels in a patient with familial pure gonadal dysgenesis.

There is experimental evidence that a portion of follicle-stimulating hormone (FSH) secretion is independent of hypothalamic influences. A 29 year old woman with familial pure gonadal dysgenesis developed myelodysplastic syndrome. Endocrine investigations showed discrepancy between serum FSH and luteinizing hormone (LH) levels. FSH levels remained elevated while LH levels decreased. The FSH to LH ratio was 10 (normal 2-2.5). The fall in LH is likely to be due to factor(s) involved directly and specifically in LH synthesis and release. Exogenous LH releasing hormone administration as well as hormonal replacement treatment increased LH levels. The FSH to LH ratio decreased to 7. This case supports the hypothesis of differential regulation of FSH and LH, and that FSH secretion is at least partly autonomous.     

Effects of Antisense Oligodeoxynucleotide to Follicle-stimulating Hormone Receptor on the Expression of Proliferating Cell Nuclear Antigen and Vascular Endothelial Growth Factor in Primary Culture Cells Derived from Human Ovarian Mucinous Cystadenocarcinoma

The effects of antisense oligodeoxynucleotide （antisense ODN） to follicle-stimulating hormone receptor （FSHR） and follicle-stimulating hormone （FSH） on the expression of proliferating cell nuclear antigen （PCNA） and vascular endothelial growth factor （VEGF） were studied in primary culture cells derived from human ovarian mucinous cystadenocarcinoma （OMC）. The primary （）MC cells were cultured with the enzyme digestion method, and the expression of pan Keratin protein and FSHR mRNA was detected for identification of the cells. OMC cells were co-cultured with antisense ODN, nonsense ODN and FSH with different concentrations for 48 h and 72 h. The expression of PCNA and VEGF was detected by using SP immunohistochemistry. Compared with that in the control group, the PCNA and VEGF expression was increased obviously in FSH groups （P〈0.05 or P〈 0.01）, while decreased significantly in antisense ODN groups （P〈0.05 or P〈 0.01） and unchanged in nonsense ODN groups, respectively. Meanwhile, antisense ODN could antagonize the increased expression of PCNA and VEGF caused by FSH significantly （P〈0.01）. It was suggested that FSH might promote the development of OMC to some extent. Antisense ODN could inhibit the proliferative activity of OMC cells and the promoting proliferative activity enhanced by FSH.     

Effects of Antisense Oligodeoxynucleotide to Follicle-stimulating Hormone Receptor on the Cell Proliferation and Apoptosis in Cells Derived from Human Ovarian Mucinous Cystadenocarcinoma in Vitro

The human ovarian mucinous cystadenocarcinoma (hOMC) cells were co-cultured with antisense oligodeoxynucleotide (antisense ODN), nonsense ODN, and follicle-stimulating hormone (FSH) at different concentrations for the purpose of observing the effects of antisense ODN to FSH receptor (FSHR) on the proliferation and apoptosis of cultured hOMC cells in vitro. The inhibitory rates of growth were measured by using MTT method on the 2nd, 4th, 6th, 8th and 10th days after the interference of antisense ODN, nonsense ODN, and FSH, respectively. The apoptotic rates and the cell cycles were determined by means of flow cytometry, the apoptosis indexes were detected by us-ing TUNEL, and the expression of caspase-3 was measured by using SP immunohistochemistry. Compared with that in the control group, the proliferative activity of hOMC cells was increased ob-viously in FSH groups (P<0.05 or P<0.01), decreased distinctly in antisense ODN groups (P<0.05 or P<0.01), and unchanged in nonsense ODN groups, respectively. Meanwhile, antisense ODN could significantly antagonize the FSH-promoted cell proliferative activity (P<0.01). Compared with those in the control group, the apoptotic rates and the expression of caspase-3 were dramatically increased in the mid- and high-dose antisense ODN groups (P<0.05 or P<0.01), while the number of cells in G1/G0 phase was significantly decreased and that in S phase distinctly increased (P<0.01). There was no change in nonsense ODN groups (P>0.05). It was suggested that FSH may improve the develop-ment of hOMC cells. However, antisense ODN could inhibit proliferative activity and the FSH-promoted proliferative activity in hOMC cells, at the same time, antisense ODN could inhibit hOMC cell growth by inducing apoptosis.     

软脂酸增加血管内皮细胞Toll样受体4表达及活化TLR4炎症信号通路

目的研究饱和脂肪酸软脂酸对血管内皮细胞Toll样受体4（TLR4）表达及信号转导的影响。方法采用软脂酸处理猪髋
动脉内皮细胞（PIECs）,实时荧光定量PCR检测PIECs TLR4 mRNA表达,蛋白印迹法检测PIECs TLR4及IκBα蛋白表达,流式
细胞仪检测PIECs细胞表面TLR4蛋白水平,酶联免疫吸附测定法检测细胞培养液TNF-α及IL-6水平。结果与对照组比较,
软脂酸组PIECs TLR4 mRNA及蛋白表达显著升高（TLR4 mRNA：4.73±0.61 vs 1.25±0.90,P<0.05;TLR4 蛋白：5.79±0.05 vs
4.07±0.31,P<0.05）,PIECs细胞表面TLR4蛋白水平显著增加（38.070±3.907 vs 29.390±1.072,P<0.05）;软脂酸组PIECs IκBα蛋
白表达显著降低（2.04±0.22 vs 3.98±0.18,P<0.05）,细胞培养液TNF-α及IL-6水平显著升高（TNF-α：2.52±0.30 vs 1.38±0.26,P<
0.05;IL-6：3.28±0.32 vs 1.44±0.28,P<0.05）。结论软脂酸在诱导血管内皮细胞TLR4表达的同时活化TLR4炎症信号通路。
     

卵泡刺激素(FSH)对卵巢癌HEY细胞系Notch通路及转录因子Snail表达的影响

 目的  研究卵泡刺激素(follicle stimulating hormone,FSH)体外对卵巢癌HEY细胞系Notch通路上Notch1胞内段蛋白(N1ICD)、转录因子CSL与靶基因表达以及对转录因子Snail家族的调节作用,以探讨FSH促进卵巢癌侵袭的可能机制。 方法  体外培养卵巢癌HEY细胞系,设PBS对照组和FSH 40mU/mL实验组,改良的Boyden小室检测两组细胞侵袭力,Western blot检测两组N1ICD、CSL、Snail、Slug蛋白表达,RT PCR检测HEY 1、HEY 2、HES 1 mRNA表达。结果  FSH 40 mIU/mL可显著增强卵巢癌HEY细胞系的侵袭能力(P=0.001)。相比于对照组,实验组FSH 40 mIU/mL作用24 h后N1ICD和CSL蛋白表达显著上升(P=0.001,P=0.028)。FSH 40 mIU/mL作用后18~24 h HEY 1 mRNA的表达增加,作用24 h时HEY 2 mRNA 的表达增加。而FSH作用后HES 1 mRNA水平无明显变化。FSH 40 mIU/mL作用24 h后,相比于对照组 Snail 蛋白表达显著上升(P=0.009),而Slug蛋白的表达无显著变化(P>0.05)。结论  FSH可能激活Notch1信号通路和Snail转录因子,进而促进卵巢癌细胞的侵袭能力。     

雌激素受体在人类多激素型垂体腺瘤中的表达

目的 研究人类多激素型垂体腺瘤中雌激素受体(ER)在分泌不同类型激素的垂体腺瘤细胞中的表达,探讨多激素型垂体腺瘤中分泌特定类型激素的垂体腺瘤细胞与ER阳性细胞的关系。方法 采用免疫组织化学双标法检测33例多激素型垂体腺瘤标本中垂体激素合并ER表达的情况,即分别检测每1例标本中PRL ER、GH ER、ACTH ER、PSH ER、FSH ER和LH ER的表达情况。结果 33例多激素型垂体腺瘤标本中有22例(66．67％)表达ER,其中：PRL ER双标染色阳性标本10例、IH ER双标染色阳性标本9例、FSH ER双标染色阳性标本7例、GH ER双标染色阳性标本2例,33例标本的ACTH ER和TSH ER的双标染色均为阴性。结论 多激素型垂体腺瘤中,分泌：PRL、IH或FSH的垂体腺瘤细胞可表达ER;分泌ACTH或TSH的垂体腺瘤细胞不表达ER;分泌GH的垂体腺瘤细胞是否表达ER可能与该垂体腺瘤是否同时分泌PRL有关。ER很可能在多激素型垂体腺瘤PRL、IH及FSH腺瘤细胞的发生、发展过程中发挥作用。     

卵泡刺激素对体外培养大鼠颗粒细胞凋亡的影响

目的：探讨FSH对体外培养大鼠颗粒细胞凋亡的影响。方法：分别将新鲜卵巢和冻融卵巢卵泡颗粒细胞分离出来，体外培养4d，对比培养液中添加FSH与未添加FSH条件下的颗粒细胞凋亡率。结果：培养液中添加FSH后，冻融颗粒细胞凋亡率降低。结论：培养液中添加10ng／ml FSH，可减少从新鲜和冻融卵泡分离的颗粒细胞在体外培养过程中发生凋亡的几率。     

FSH/LH在AMH正常人群中评估卵巢低反应的价值

目的 研究在抗苗勒管激素（anti-müllerian hormone,AMH）正常人群中,促卵泡激素与黄体生成素比值（follicle-stimulating hormone/luteinizing hormone,FSH/LH）升高是否与卵巢低反应的发生相关,以期找到诊断临界值。 方法 分析我院生殖中心进行体外受精-胚胎移植（in vitro fertilization-embryo transfer,IVF-ET）助孕的周期4 363个,通过平滑曲线拟合分析在1.1＜AMH＜6 μg/L人群中FSH/LH与卵巢低反应发生的关系,寻找阈值或饱和点,根据找到的阈值或饱和点分组,比较两组人群特征、周期信息、周期结局,分析FSH/LH升高与卵巢低反应及周期结局的变化。 结果 在AMH正常人群中,通过平滑曲线拟合发现,随FSH/LH水平增加,卵巢低反应发生增加,但通过阈值与饱和分析找出最佳饱和点为3.3。通过FSH/LH≥3.3组与FSH/LH＜3.3组比较,年龄、体重指数、FSH、促性腺激素（gonadotropins,Gn）启动剂量、Gn总剂量均显著升高;AMH、窦卵泡数（antral follicle counting,AFC）、基础LH、雌激素（estrogen,E2）、获卵总数、可移植胚胎率、扳机日E2、孕激素（progesterone,P）水平均降低（P＜0.05）。 结论 在控制性超促排卵过程中,AMH是预测卵巢反应性的可靠指标。在AMH正常人群中,随FSH/LH升高卵巢低反应的发生率增加,FSH/LH=3.3为饱和值,但预测卵巢低反应发生的准确率仅有23%, FSH/LH升高的人群中,FSH升高及LH降低都与卵巢低反应发生相关。FSH/LH升高使最终获卵率及可移植胚胎率降低,但不影响优质胚胎率及最终的妊娠率。     

FSH和IGF-1对大鼠卵巢细胞微囊化前后的影响

目的：观察微囊化卵巢细胞的形态变化及卵泡刺激素（FSH）和胰岛素样生长因子-1（IGF-1）对其增殖功能的影响,探讨微囊化卵巢细胞作为内源性雌激素来源的可行性。方法：分离21 d龄雌性SD大鼠卵巢细胞,将培养至第一代的卵巢细胞微囊化包裹后接种于96孔板,加入含不同浓度的FSH和IGF-1（浓度分别为0、6.3、12.5、25、50、100 ng/mL）的无血清培养液;另将正常培养的卵巢细胞培养24 h后换无血清培养液进行相同的处理,培养44 h后,加入WST-1试剂,继续孵育4 h。全自动酶标仪测定光密度。结果：卵巢细胞在微囊内呈圆形均匀分布,48 h后形态无显著变化。FSH对培养的卵巢细胞和微囊化的卵巢细胞的增殖刺激作用均呈剂量依赖性,卵巢细胞的增殖刺激作用在FSH 50 ng/mL时,达到最高峰（P〈0.05）;对微囊化卵巢细胞的刺激作用在FSH 100 ng/mL时,细胞的增殖活性达到最高峰（P〈0.05）;IGF-1对卵巢细胞的刺激增殖作用在低剂量组不显著（6.3、12.5、25 ng/mL,P〉0.05）,在高剂量组（50、100ng/mL）对卵巢细胞的刺激作用与对照组相比差异有统计学意义（P〈0.05,P〈0.01）。IGF-1对微囊化卵巢细胞的刺激增殖作用呈剂量依赖性（P〈0.05）,在IGF-1 50 ng/mL和100 ng/mL组,微囊化卵巢细胞的OD值与对照组相比,差异有统计学意义（P〈0.01）。结论：卵巢细胞在微囊化后,可以在微囊内存活并保持其生物学活性,并且可以对外源性的FSH和IGF-1的刺激产生反应。     

Effects of Antisense Oligodeoxynucleotide to Follicle-stimulating Hormone Receptor on the Expression of Proliferating Cell Nuclear Antigen and Vascular Endothelial Growth Factor in Primary Culture Cells Derived from Human Ovarian Mucinous Cystadenocarcino

The effects of antisense oligodeoxynucleotide (antisense ODN) to follicle-stimulating hormone receptor (FSHR) and follicle-stimulating hormone (FSH) on the expression of proliferating cell nuclear antigen (PCNA) and vascular endothelial growth factor (VEGF) were studied in primary culture cells derived from human ovarian mucinous cystadenocarcinoma (OMC). The prlmary OMC cells were cultured with the enzyme digestion method, and the expression of pan Keratin protein and FSHR mRNA was detected for identification of the cells. OMC cells were co-cultured with antisense ODN, nonsense ODN and FSH with different concentrations for 48 h and 72 h. The expression of PCNA and VEGF was detected by using SP immunohistochemistry. Compared with that in the control group, the PCNA and VEGF expression was increased obviously in FSH groups (P＜0.05 or P＜ 0.01), while decreased significantly in antisense ODN groups (P＜0. 05 or P＜0.01) and unchanged in nonsense ODN groups, respectively. Meanwhile, antisense ODN could antagonize the increased expression of PCNA and VEGF caused by FSH significantly (P＜0.01). It was suggested that FSH might promotethe development of OMC to some extent. Antisense ODN could inhibit the proliferative activity of OMC cells and the promoting proliferative activity enhanced by FSH.     

促卵泡生成素,黄体生成素和胰岛素对多囊卵巢病人颗粒细胞甾 …

目的 研究在促卵泡生成素（ＦＳＨ）促黄体生成素（ＬＨ）和胰岛素（ＩＮＳ）刺激作用下多囊卵巢综合征（ＰＣＯＳ）病人与正常卵巢的颗粒细胞甾体激素产生的异同。方法 对１８例月经周期正常妇女卵泡期的３３个卵泡和８例ＰＣＯＳ的１００个肺泡的颗粒细胞行体外培养,利用放射免疫法测定培养液中的甾体激素水平,比较了两组颗粒细胞在ＦＳＨ,ＬＨ和ＩＮＳ刺激作用下下雌二醇（Ｅ２）和孕酮（Ｐ）的养液中的甾体激素水平,比较了     

Reactivity of human preformed natural antibodies with various porcine pancreatic cells

Pancreatic islet transplantation is a promisingtherapeutic approach for the treatment of insulin- de-pendent diabetes mellitus( IDDM) [1] .Because of se-vere shortage of human organs,the pig appears to bethe most likely donor species for xenotransplantationinto humans for a variety of reasons[2 ] .However,successful organ xenotransplantation requireselimina-tion of preformed natural antibodies( PNAbs) re-sponsible forhyperacute graftrejection.Itis notwellknown whether human PNAbs to the po…     

无精症睾丸病理与血清FSH水平的关系及对不育诊治的意义

目的:探讨无精症患者睾丸活检与血清FSH水平关系,及其在无精症诊断、治疗中的意义.方法:用放免法测定67例无精症患者血清FSH水平,并取睾丸活检,对其进行病理学分类.比较不同病理学类型的无精症患者血清FSH水平.结果:唯支持细胞综合征患者血清FSH水平显著高于正常组、生精功能低下组及生精阻滞组.而生精功能低下、生精阻滞组血清FSH水平与正常组间无差异.结论:无精症患者血清FSH水平对于诊断难支持细胞综合征具有一定价值.对于其他类型的生精功能受损,测定血清FSH水平预测作用.睾丸活检对无精症的诊治具有重要的指导意义.     

不同年龄段患者基础FSH/LH比值对IVF/ICSI-ET临床结局的预测价值

目的 探讨患者基础卵泡刺激素/基础黄体生成素(FSH/LH)比值对体外授精/卵胞浆内单精子注射-胚胎移植(IVF/ICSI-ET)过程中获卵总数、优质胚胎的获得和临床结局的预测价值.方法 回顾性分析因单纯输卵管因素或男方因素行IVF/ICSI-ET(长方案)助孕并行新鲜胚胎移植的1629例患者的病历资料,根据患者的年龄和FSH/LH比值分为A、B、C共3组,A组:年龄<30岁(A1亚组:FSH/LH<2,n=452;A2亚组:FSH/LH≥2,n=144);B组:年龄30~35岁(B1亚组:FSH/LH<2,n=551;B2亚组:FSH/LH≥2,n=240);C组:年龄36~40岁(C1亚组:FSH/LH<2,n=148;C2亚组:FSH/LH≥2,n=94).比较各年龄段亚组患者控制性超促排卵(COH)结果以及IVF结局.结果 A组中,A1亚组的促性腺激素(Gn)用量明显低于A2亚组,差异有统计学意义(P<0.05);A1亚组的获卵数和优质胚胎数明显高于A2亚组,差异有统计学意义(P<0.05).B组中,B1亚组的获卵数明显低于B2亚组,差异有统计学意义(P<0.05).C组中,C1亚组和C2亚组在获卵数和优质胚胎数之间差异也具有统计学意义(P<0.05).A、B、C 3组各亚组在患者的妊娠率、流产率差异均无统计学意义.结论 基础FSH/LH比值可以作为判断不同年龄段患者COH结果的重要指标,但却不能单独预测IVF/ICSI-ET临床结局.