宫内节育器对大鼠子宫内膜血管内皮生长因子及其受体表达影响的研究

目的:研究置固定式铜宫内节育器(FCu-IUD)和固定式吲哚美辛铜宫内节育器(FICu-IUD)的大鼠子宫内膜组织中血管内皮生长因子(VEGF)及其受体Flk-1的表达。方法:采用蛋白印迹法(Western blotting)检测FCu-IUD组、FICu-IUD组大鼠子宫内膜组织VEGF及Flk-1的表达,逆转录聚合酶联反应(RT-PCR)检测各组VEGF mRNA的表达。以未置器一侧作为对照。结果:大鼠置器侧与对照侧子宫内膜均可检测到与VEGF121和VEGF165两种VEGF亚型相对应的mRNA与蛋白表达。FCu-IUD组大鼠置器侧子宫内膜VEGF121及VEGF165表达在mRNA水平和蛋白水平均显著升高,与对照侧比较差异有统计学意义(P<0.001,P<0.05)。置器侧Flk-1蛋白的表达明显高于对照侧(P<0·01)。FICu-IUD组置器侧与对照侧比较,VEGF、Flk-1蛋白及VEGF mRNA表达差异均无显著性。结论:Cu-IUD引起的子宫异常出血可能与VEGF的增加有关,吲哚美辛减少Cu-IUD引起的子宫异常出血的作用可能与通过抑制前列腺素合成从而使VEGF生成减少有关。     

原因不明不孕患者子宫内膜胰岛素样生长因子Ⅱ及其受体mRNA的表达及其与性激素水平的相关性

目的探讨胰岛素样生长因子Ⅱ(IGF-Ⅱ)及其Ⅰ型受体(IGF-ⅠR)、Ⅱ型受体(IGF-ⅡR) mRNA在原因不明不孕患者黄体中期子宫内膜的表达,及其与性激素水平变化的相关性.方法采用原位杂交、逆转录PCR(RT-PCR)技术,检测38例原因不明不孕患者(研究组)和20例正常育龄女性志愿者或因男方因素不孕患者(对照组)黄体中期子宫内膜IGF-Ⅱ、IGF-ⅠR、IGF-ⅡR mRNA的表达;放射免疫法测定同期血清雌二醇、孕酮水平.结果黄体中期子宫内膜间质细胞胞浆内,可见IGF-Ⅱ、IGF-ⅡR蓝色颗粒弥散分布,IGF-ⅠR 则主要在上皮细胞中分布.研究组IGF-Ⅱ mRNA的表达(灰度值,下同)为0.71±0.34,对照组为0.96±0.34,两组比较,差异有显著性(P<0.05);研究组IGF-ⅠR mRNA的表达为0.62±0.28,显著低于对照组的0.93±0.51(P<0.05);研究组IGF-ⅡR mRNA的表达为0.49±0.27,也显著低于对照组的0.73±0.36(P<0.05).研究组黄体中期孕酮水平为(34±15)nmol/L,显著低于对照组(53±17)nmol/L(P<0.01), 而血清雌二醇水平与对照组比较,差异无显著性(P>0.05).研究组与对照组IGF-Ⅱ mRNA与IGF-ⅠR、IGF-ⅡR mRNA的表达呈正相关;两组妇女孕酮与IGF-Ⅱ、IGF-ⅠR mRNA表达水平呈正相关;对照组孕酮与IGF-ⅡR mRNA表达水平呈正相关.结论原因不明不孕患者黄体中期孕酮不足,可能影响子宫内膜IGF-Ⅱ、IGF-ⅠR、IGF-ⅡR mRNA的表达,并可能是导致不孕的病因之一.     

经宫颈子宫内膜电切术后微血管密度、血管内皮因子及其受体表达变化的研究

目的 研究微血管密度(microvessel density,MVD)、血管内皮生长因子(vascular endothelial growth factor,VEGF)及其受体(vascular endothelial growth factor receptor,KDR)在单纯增生、腺肌病患者经宫颈子宫内膜电切手术(transcervical resection of endometrium,TCRE)前后表达的变化，探讨子宫内膜电切手术治疗子宫异常出血的分子学机制。方法 功能失调性子宫出血患者诊刮病理为单纯增生内膜，于刮宫后12～15d行电切手术，取电切内膜标本10例、子宫腺肌病增殖中晚期的患者电切子宫内膜10例为术前实验组；功血患者于TCRE术后6～18个月取内膜8例、子宫腺肌病患者于TCRE术后6～18个月取内膜7例为术后实验组；正常对照选取增殖中晚期子宫内膜8例为对照组。采用免疫组织化学LSAB法检测组织中MVD、VEGF及其受体KDR的表达，并抗第八因子相关抗原抗体标记间质微血管并进行微血管计数、以计算机图像分析获得VEGF、KDR值。结果 单纯增生及腺肌病之子宫内膜其MVD、VEGF及KDR的表达在TCRE术前与正常对照组和术后的表达比较差异均有显著性；单纯增生和腺肌病TCRE术后的子宫内膜MVD、VEGF及KDR的表达与正常对照组比较差异无显著性。单纯增生与腺肌病之子宫内膜MVD、VEGF及KDR表达比较无明显统计学意义，但腺肌病子宫内膜MVD、VEGF及KDR的均值比单纯增生高。结论 子宫内膜电切术对子宫内膜MVD、VEGF及KDR的表达起降调节，为子宫内膜电切术治疗子宫异常出血提供理论依据。对于腺肌症患者术后辅以抗血管药物可能有助于抑制复发。     

手术后口服小剂量米非司酮治疗子宫内膜异位症的近期疗效和副作用的初步观察

目的探讨保留生育功能或保留卵巢功能手术后服用小剂量米非司酮治疗子宫内膜异位症(内异症)的临床疗效、副反应及对生殖激素水平和骨代谢的影响.方法米非司酮组31例,予米非司酮10 mg口服,每日1次;达那唑组30例,予达那唑200 mg口服,每日2～3次,均连续用药3个月.观察两组症状、体征改善情况,用药后的副反应及用药3个月时血清生殖激素水平和骨代谢生化指标的变化.结果两组症状、体征缓解率相似.米非司酮组潮热、阴道出血、腰背疼痛、体重增加、痤疮等副反应发生率显著低于达那唑组(P<0.05).用药3个月时,米非司酮组血清促卵泡激素和黄体生成激素与达那唑组比较,差异无显著性(P>0.05);米非司酮组雌二醇(E2)为(204.9±45.3) pmol/L,保持在卵泡期水平,达那唑组为(94.3±33.0) pmol/L,为绝经后水平,两组比较,差异有极显著性(P<0.01).停药13～15 d,米非司酮组E2水平为(1 221.6±384.2) pmol/L,较用药前排卵期下降,但差异无显著性(P>0.05),达那唑组为(815.1±376.0) pmol/L,明显下降(P<0.05).停药当月黄体高峰期孕酮水平,米非司酮组为(33.1±5.6) nmol/L,与治疗前相似,而达那唑组为(27.4±4.9) nmol/L,显著降低(P<0.01),两组间差异有显著性(P<0.05).用药3个月时,米非司酮组尿脱氧吡啶啉/尿肌酐(UDpd/Cr)、血碱性磷酸酶(AKP)和骨钙素(BGP)与用药前比较,差异均无显著性(P>0.05);达那唑组血AKP为(54.0±10.7) U/L,BGP为(7.7±1.9) μg/L,较用药前明显升高(P<0.01),而UDpd/Cr水平无显著变化(P>0.05).米非司酮组停药后基础体温上升和月经复潮时间较达那唑组为短.结论手术后加用米非司酮可明显改善患者症状和体征,疗效与达那唑类似,但副反应明显减少.10 mg/d米非司酮3个月治疗可使血清E2保持在卵泡期水平,且对骨代谢无明显影响.     

血管内皮生长因子在卵巢过度刺激综合征发病机理中作用的初步研究

目的通过对卵巢过度刺激综合征(OHSS)患者血管内皮生长因子(VEGF)水平和表达的测定,初步探讨VEGF在OHSS发病机理中的作用.方法采用酶联免疫吸附试验方法对14例中、重度OHSS患者(OHSS组)和同期13例非OHSS患者(对照组)取卵日卵泡液、人绒毛膜促性腺激素(hCG)注射日、胚胎移植(ET)日血清VEGF水平进行测定;采用免疫组织化学方法和逆转录聚合酶链反应技术检测取卵日卵巢黄素化颗粒细胞VEGF及其mRNA的表达.结果 OHSS组和对照组的卵泡液、hCG注射日血清VEGF水平分别为(1 257.2±648.0)与(1 745.1±802.4) ng/L,(250.1±109.5)与(196.7±81.7) ng/L,两组比较,差异无显著性(P>0.05);OHSS组ET日血清VEGF水平及注射hCG后血清VEGF水平变化程度,显著高于对照组[分别为(342.9±158.5)与(222.2±84.6) ng/L,(92.8±106.7)与(25.9±21.8) ng/L,P<0.05].OHSS组取卵日卵巢黄素化颗粒细胞VEGF mRNA的表达,显著高于对照组,VEGF/β肌动蛋白比值分别为1.50±0.60、0.96±0.56(P<0.05).两组取卵日卵巢黄素化颗粒细胞有较活跃的VEGF表达.结论 VEGF在OHSS的发病机理中可能起一定的作用.     

VEGF及其受体KDR在子宫内膜异位症中的表达及意义

目的:通过检测血管内皮生长因子(VEGF)、激酶插入区受体(KDR)在子宫内膜异位症(EMs)患者异位内膜、在位内膜及血清和腹腔液中的表达,探讨VEGF及KDR在EMs发病机制中的作用。方法:应用免疫组化SP法检测并比较EMs异位内膜、在位内膜及对照组子宫内膜组织中VEGF及KDR的表达水平。应用酶联免疫吸附法(ELISA)检测并比较EMs组及对照组血清和腹腔液中VEGF及KDR的含量。结果:VEGF和KDR在EMs组异位内膜的表达明显高于在位内膜和正常内膜组(P<0.05),VEGF和KDR在在位内膜的表达明显高于正常对照组(P<0.05),且两者在EMs异位内膜的表达呈正相关(r=0.971,P<0.05)。EMs患者血清和腹腔液中VEGF及KDR含量明显高于对照组(P<0.01),临床分期较早(Ⅰ～Ⅱ)的EMs患者的血清和腹腔液中VEGF及KDR水平明显高于临床分期较晚(Ⅲ～Ⅳ)的患者(P<0.05)。EMs组腹腔液中VEGF及KDR浓度显著高于其血清中浓度(P<0.05)。结论:VEGF及KDR可能在EMs发生、发展过程中发挥了重要的作用,检测血清VEGF及KDR水平为EMs的早期诊断提供了一种新的可能方法,也为通过VEGF-VEGFR双靶向阻断血管来治疗EMs奠定了理论基础。     

血管内皮生长因子及其受体在月经周期子宫内膜中的表达

目的　检测血管内皮生长因子 (VEGF)及其受体fms样酪氨酸激酶 (flt 1)、含插入区的激酶受体 (KDR)在正常子宫内膜中的表达 ,探讨其在子宫内膜血管生成及内膜生长分化中的作用。方法　采用免疫组织化学及原位杂交方法 ,对 5 0例妇女正常月经周期子宫内膜中VEGF、flt 1、KDR蛋白质及mRNA进行检测 ,用蛋白印迹法分析子宫内膜中VEGF亚型 ,用内皮细胞标志物Ⅷ因子标记内膜微血管密度。结果　VEGF、flt 1、KDR蛋白质及mRNA主要分布在正常子宫内膜血管内皮细胞及腺上皮细胞 ,VEGF、flt 1在分泌中期、月经期的表达较强 ,KDR在增生中期即达高峰 ,三者在增生早期的表达均最低。分泌期血管壁面积及血管腔面积大于增殖期 (P <0 0 5 ) ,血管密度无明显周期性改变 (P >0 0 5 )。正常子宫内膜组织 4种VEGF蛋白亚型中 ,以VEGF12 1、VEGF16 5为主 ,其免疫条带积分吸光度明显高于VEGF189、VEGF2 0 6 (P <0 0 5 )。结论　VEGF、flt 1、KDR蛋白质及mRNA在正常子宫内膜中的表达呈明显周期依赖性 ,分泌期表达增强提示其可能参与胚胎着床 ,月经期表达增加可能与此期子宫内膜脱落缺氧有关     

血管内皮生长因子在预测卵巢过度刺激综合征中的作用

目的探讨在体外受精-胚胎移植(IVF-ET)周期中血清及卵泡液中血管内皮生长因子(VEGF)水平的变化,及其在预测卵巢过度刺激综合征(OHSS)中的作用.方法收集42例行IVF-ET患者的血清和卵泡液标本,采用定量酶联免疫方法检测血清、卵泡液中VEGF水平,按是否发生OHSS分为OHSS组(10例)及对照组(32例),对两组临床资料、性激素和VEGF水平进行回顾性相关分析.结果 OHSS组血清VEGF水平在月经周期的各个时期均高于对照组,但差异无显著性(P>0.05).OHSS组卵泡液VEGF水平为(1 487.7±365.8) ng/L,较对照组的(1 025.8±474.7) ng/L明显升高;取卵时卵泡数目增多,OHSS组为(10.0±5.9)个,对照组为(6.1±2.3)个;基础黄体生成素(LH),人绒毛膜促性腺激素(hCG)、注射日雌二醇(E2)水平均比对照组高,差异有显著性(P<0.05).结论 OHSS组卵泡液VEGF水平明显高于对照组,提示VEGF可能参与了OHSS的发病;卵泡液VEGF水平可以作为预测OHSS发生的检测指标.     

白血病抑制因子在早期妊娠、先兆流产及难免流产患者蜕膜组织中表达的研究

目的探讨白血病抑制因子(LIF)在蜕膜组织中的表达,及其与早期妊娠、流产的关系. 方法采用放射免疫方法检测正常早孕妇女(正常早孕组)、先兆流产妇女(先兆流产组)及难免流产妇女(难免流产组)的血清孕酮及人绒毛膜促性腺激素(hCG)水平,并采用逆转录-聚合酶链反应(RT-PCR)技术对3组孕妇蜕膜组织中LIF-mRNA的表达进行定量分析.结果 (1)孕酮及hCG水平在3组孕妇间的比较正常早孕组孕妇血清中孕酮、hCG水平分别为(91.5±27.2) nmol/L、(69.9±14.9) kU/L,先兆流产组孕妇分别为(88.4±24.7) nmol/L、(57.6±11.2) kU/L,两组孕妇血清孕酮、hCG水平比较,差异均无显著性(P>0.05);而难免流产组孕妇血清孕酮、hCG水平分别为(33.1±19.6) nmol/L、(10.3±3.2) kU/L,与前两组分别两两比较,差异均有极显著性(P<0.01).(2)LIF-mRNA平均相对含量在3组孕妇间的比较正常早孕组孕妇为2.10±0.32;先兆流产组孕妇为1.92±0.20;难免流产组孕妇为0.70±0.06.正常早孕组与先兆流产组比较,差异无显著性(P>0.05).而难免流产组与前两组分别行两两比较,差异均有显著性(P<0.05).结论 LIF-mRNA在早期妊娠蜕膜组织中的表达量降低,可能是导致hCG及孕酮分泌下降,最终造成难免流产的原因之一.     

单核细胞趋化蛋白-1在子宫内膜异位症患者子宫内膜组织中的表达

目的检测单核细胞趋化蛋白-1（MCP-1）在子宫内膜异位症(内异症)患者子宫内膜组织中的表达情况，并探讨其与内异症发病的关系。方法内异症患者30例作为研究组，非内异症患者20例作为对照组。用半定量逆转录聚合酶链反应（RT-PCR）方法检测两组子宫内膜组织中MCP-1信使核糖核酸(mRNA)的阳性表达率及相对水平。结果研究组26例(87%)内膜组织中可检测到MCP-1mRNA的表达，其中19例为强阳性表达，但其表达强度与内异症的严重程度无明显相关性。对照组15例(75%)内膜组织中可检测到MCP-1mRNA表达，仅3例为强阳性表达。研究组内膜组织中MCP-1mRNA的表达强度高于对照组(两组相对表达强度分别为0.746±0.345及0.460±0.341，P<0.05)。结论内异症患者的子宫内膜组织中MCP-1mRNA的表达增强，MCP-1对内异症的发病有一定促进作用。     

VEGF在子宫内膜异位症发病机制中的作用

目的:探讨血管内皮生长因子(Vascular endothelial growth factor,VEGF)在子宫内膜异位症(Endometriosis,EMs)组织中的表达及其在EMs发生发展中的作用。方法:采用ELISA法检测30例EMs患者(研究组)和16例子宫肌瘤患者(对照组)血清和腹腔液中的VEGF水平,免疫组化和RT-PCR检测并比较VEGF蛋白、VEGF mRNA在EMs异位内膜、在位内膜和对照组正常内膜的表达及其相关性。结果:EMs患者腹腔液、异位内膜、在位内膜VEGF蛋白及mRNA均明显高于对照组(P<0.05),但与EMs临床分期无关。EMs患者血清VEGF与对照组相比无明显的差异(P>0.05)。EMs患者异位内膜和在位内膜的微血管密度(MVD)明显高于对照组内膜(P<0.05)。结论:VEGF在EMs患者的高表达,说明其通过促血管生成在该病发生发展中起着重要作用。     

Expression of vascular endothelial growth factor in adenocarcinomas of the uterine cervix and its relation to angiogenesis and p53 and c-erbB-2 protein expression

OBJECTIVE: The purpose of this study was to evaluate vascular endothelial growth factor (VEGF) expression in adenocarcinomas of the uterine cervix and its correlation with clinicopathologic features, angiogenesis, and expression of p53 and c-erbB-2 proteins. METHODS: Thirty-seven cases of FIGO clinical stage I and II adenocarcinoma of the uterine cervix were examined by immunohistochemical studies with anti-VEGF, anti-CD34, anti-p53, and anti-c-erbB-2 antibodies. Computerized image analysis was used to evaluate microvessel density (MVD). RESULTS: Thirty-one tumors (83.8%) were classified as VEGF positive. Six tumors (16.2%) showed p53 protein expression while 11 tumors (29.7%) expressed the c-erbB-2 protein. MVD ranged from 13.3 to 44.8, with a median value of 25.5 (26.9 +/- 7.5). Tumors expressing VEGF had a significantly higher MVD than those that did not express VEGF (P < 0.05). VEGF expression was significantly associated with c-erbB-2 protein expression (P < 0.05). The spatial distributions of both VEGF expression and c-erbB-2 expression were similar in tumor tissues. In univariate log-rank analysis, stage (P = 0.0250), lymphovascular space invasion (P = 0.0156), and MVD (P = 0.0360) were associated with shortened survival. CONCLUSION: VEGF expression plays a role in promoting angiogenesis in cervical adenocarcinomas and c-erbB-2 is likely to be involved in the up-regulation of VEGF expression.     

子宫内膜增生过长和子宫内膜样腺癌中VEGF 和TSP-1表达与血管新生研究

目的探讨子宫内膜增生过长及子宫内膜样腺癌中血管内皮生长因子(VEGF)和血小板反应素1(TSP-1)表达与血管新生的关系以及对子宫内膜样腺癌的发生、发展中的作用.方法采用免疫组织化学方法分别检测子宫内膜正常(12例)、增生过长(13例)、不典型增生(18例)及子宫内膜样腺癌(50例)中微血管密度(MVD)、VEGF及TSP-1表达情况.结果子宫内膜不典型增生和内膜样腺癌中MVD明显大于内膜正常及增生过长者(P＜0.05),子宫内膜样腺癌IA期与不典型增生二者的MVD差异无显著性(P＞0.05),而与正常内膜和增生过长者差异有显著性(P＜0.05);VEGF表达与上述不同内膜病变中MVD呈正相关(r=0.843,P=0.000 1),而TSP-1表达仅在子宫内膜样腺癌中分别与MVD和VEGF呈负相关趋势(r=-0.233,P=0.1041;r=-0.235,P=0.100 3);在子宫内膜样腺癌中TSP-1间质高表达且具有异质性.结论子宫内膜不典型增生和子宫内膜样腺癌中VEGF对血管新生起正向调节作用;TSP-1在部分腺癌患者中表达增强,但其负向调节作用较弱,可能会使血管新生开关平衡失调,与子宫内膜样腺癌发生及发展有关.     

The association between vascular endothelial growth factor, microvessel density and clinicopathological features in invasive cervical cancer

OBJECTIVE: The aim of this study was to analyse the vascular endothelial growth factor (VEGF) expression in a series of cervical carcinomas and to compare the results with the microvessel density (MVD) and clinicopathological features. STUDY DESIGN: The immunoreactivity for VEGF was studied in 130 invasive cervical carcinomas and in 22 patients with a carcinoma in situ of the cervix. The results were compared with the MVD. RESULTS: Staining for VEGF of less then 50% per slide occurred in 80% of the invasive carcinomas and in 82% of the in situ carcinomas. The median MVD was 261 vv/mm(2) (range: 11-1000) in the invasive group and 146 vv/mm(2) (range: 25-536) in the in situ group. Unlike the microvessel density there was no association between VEGF expression and survival. The MVD was higher in VEGF poorer (<50%) tumours (P=0.055). Beside tumour histology (P=0.012) there were no other significant relationships between the remaining histopathological findings and VEGF expression. CONCLUSION: Tissue VEGF expression has no prognostic value in contrast with the MVD in patients with invasive cervical cancer.     

Matrix metalloproteinase-2 and -9 expression correlated with angiogenesis in human adenomyosis

BACKGROUND: Adenomyosis is an important cause of dysmenorrhea and infertility for women all over the world, however, the pathogenesis has not been completely elucidated. The purpose of the study was to investigate the role of matrix metalloproteinases (MMPs) and their relation to angiogenesis in human adenomyosis. METHODS: Adenomyotic endometrial specimens were removed by hysterectomy from 68 women with adenomyosis. The control group consisted of 26 normal endometrial specimens. Immunohistochemistry was used to demonstrate expression of MMP-2, -9, vascular endothelial growth factor (VEGF) and microvessel density (MVD). Staining intensity was analyzed by computerized image analysis system. RESULTS: In both eutopic and ectopic endometrium of adenomyosis, the expression of MMP-2, -9 as well as VEGF was significantly greater than in normal endometrium (p < 0.05). MVD was higher in ectopic endometrium than eutopic endometrium with or without adenomyosis (p < 0.05). In adenomyosis, a positive correlation was observed between VEGF expression and MMP-2 (p < 0.001, r = 0.583) as well as MMP-9 expression (p = 0.002,r = 0.490). A positive correlation was also found between MVD and MMP-2 (p < 0.001,r = 0.589) or MMP-9 expression (p < 0.001,r = 0.589). CONCLUSION: Our results indicate that the elevation of MMP-2, -9 expression may have an important role in the development of adenomyosis, probably through contributing to invasion of endometrial tissues into the myometrium and angiogenesis in adenomyotic implants.     

Relationship of hypoxia-inducible factor-1alpha expression and vascular endothelial growth factor in SKOV-3 ovarian cancer model

OBJECTIVE: To investigate the correlation of hypoxia-inducible factor (HIF)-1alpha and vascular endothelial growth factor (VEGF) or micro-vessel density (MVD). METHODS: The ovarian cancer cell line SKOV3 was transplanted into nude mice to form xenogeneic tumor. Mice were treated with rapamycin 4 mg/kg, sulindac 100 mg/(kg.d) and saline 200 microl respectively. Expression of HIF-1alpha and VEGF proteins and MVD were determined by immunohistochemistry. The mRNA of Glut1 and VEGF was studied by RT-PCR. RESULTS: The positive expression of HIF-1alpha and VEGF was moderate to strong, and MVD was high (31 +/- 8) in control group. In rapamycin treated group, the expression of HIF-1alpha was inhibited to weak positive or negative (P < 0.05), the VEGF protein and mRNA expression decreased (P < 0.05), MVD was also suppressed to a low level (13 +/- 5). There was a significant correlation between HIF-1alpha expression and VEGF expression (by Spearman's coefficient r = 0.8264, P < 0.01), and MVD (r = 0.4842, P < 0.05). CONCLUSIONS: HIF-1alpha protein can regulate VEGF protein and its mRNA expressions. It correlates with MVD in ovarian tumor tissue.