雌、孕激素对人子宫内膜Pbx2蛋白表达的影响

目的:探讨人子宫内膜腺上皮细胞和基质细胞中雌、孕激素对Pbx2蛋白表达的影响。方法:采用免疫组织化学链霉菌抗生物素蛋白-过氧化物酶(SP)连接法检测人增生期、分泌中期和蜕膜期子宫内膜中Pbx2的表达和分布;体外培养人子宫内膜基质细胞和高分化子宫内膜癌细胞Ishikawa,分别加入雌激素、孕激素、雌、孕激素联合刺激48 h,并以不加雌、孕激素的基质细胞核和Ishikawa细胞为对照组,采用免疫组织化学、免疫印迹法测定各种条件下Ishikawa细胞中Pbx2蛋白的表达。结果:①在人各期子宫内膜组织中,Pbx2表达于腺上皮细胞和基质细胞的细胞核中;Pbx2在分泌中期和蜕膜期基质细胞中的表达显著高于增生期,但在腺上皮细胞中增生期组织的表达高于分泌中期和蜕膜期,差异均具有统计学意义(P<0.05)。②Western blotting结果显示,在孕激素和雌、孕激素联合处理Ishikawa细胞组中,Pbx2的表达显著低于对照组(P<0.05),雌激素处理后Pbx2的表达与其他各组间的表达无显著性差异(P>0.05),在人子宫内膜基质细胞(ESC)中,Pbx2的表达在雌、孕激素处理各组间比较均无统计学差异(P>0.05)。结论:在人子宫内膜腺上皮Ishikawa细胞中孕激素对Pbx2的表达具有下调作用,在人子宫内膜基质细胞中,Pbx2的调控可能是非雌、孕激素依赖性的。     

Bcl-2、Bax在米非司酮治疗前后子宫内膜单纯性增生组织中的表达

目的:探讨米非司酮对子宫内膜单纯性增生的调节机制。方法:正常子宫内膜组织10例(增生期和分泌期各5例),单纯性增生子宫内膜43例,同一患者口服3个月米非司酮(10 mg/d)治疗后的子宫内膜标本43例。采用链酶菌素-生物素(S-P)免疫组化染色方法测定bcl-2、bax的表达量采用SPSS11.5统计学软件包进行数据处理,以P<0.05为有显著性差异。结果:增生期子宫内膜bcl-2阳性信号明显高于分泌期(P<0.05),单纯性增生bcl-2表达明显高于正常增生期子宫内膜(P<0.05)。与单纯性增生相比,米非司酮治疗后,子宫内膜bcl-2明显下降(P<0.05)。分泌期bax表达明显高于增生期(P<0.05),单纯性增生bax表达明显低于正常增生期子宫内膜(P<0.05)。与单纯性增生相比,米非司酮治疗后,子宫内膜bax明显增高(P<0.05)。结论:RU486增加了子宫内膜bax的表达,减少了bcl-2的表达,可能是其促进细胞凋亡,抑制子宫内膜增生的机制之一。     

Foxp3在子宫内膜、正常早孕和原因不明复发性流产患者蜕膜上的表达

目的:探讨Foxp3在增生期、分泌期内膜和正常早孕、原因不明复发性流产(URSA)患者蜕膜的表达。方法:用免疫组化法观察15例增生期子宫内膜, 12例分泌期子宫内膜, 32例正常早孕和25例URSA患者蜕膜组织Foxp3的表达与分布。结果:增生期子宫内膜无Foxp3表达,分泌期内膜和蜕膜组织均有表达;分泌期内膜Foxp3的表达显著低于正常早孕组(P<0 .01)和URSA组(P<0. 05);URSA患者蜕膜Foxp3的表达显著低于正常早孕组(P<0 .01);Foxp3表达于胞浆,正常早孕组和分泌期内膜主要表达在腺上皮细胞,URSA患者主要表达在间质细胞。结论:Foxp3在分泌期子宫内膜和蜕膜组织的表达可能在胚泡植入和早期妊娠的维持中起重要作用,其表达水平降低可能与URSA的发生有关。     

Fhit和ki-67在子宫内膜异位症的表达及意义

目的:通过观察脆性组氨酸三联体Fhit和细胞核增殖抗原ki-67在子宫内膜异位症的表达,探讨Fhit和ki-67在子宫内膜异位症发病中的作用。方法:用免疫组化SP法检测58例子宫内膜异位症(EMS)患者的异位内膜和在位内膜及15例子宫肌瘤患者子宫内膜(对照组)Fhit和ki-67的表达。结果:Fhit主要在腺上皮细胞的胞浆和胞膜表达,其表达强弱顺序为:对照组子宫内膜>EMS组在位内膜>EMS组异位内膜,EMS在位和异位内膜Fhit表达在增生期和分泌期无统计学差异(P>0.05);EMS组在位内膜的Fhit表达在临床r-AFS不同分期中无统计学差异(P>0.05),但异位内膜的Fhit表达在临床r-AFS不同分期中差异显著。ki-67主要在腺上皮细胞的胞核和胞膜表达,EMS在位内膜和对照组子宫内膜增生期ki-67表达均显著强于分泌期,EMS异位内膜腺体中ki-67表达在增生期和分泌期无统计学差异(P>0.05);EMS组异位内膜腺体ki-67表达在增生期弱于自身在位内膜(P<0.01),但分泌期强于其自身在位内膜(P<0.01)。EMS异位内膜腺体Ⅰ~Ⅱ期的ki-67表达高于Ⅲ、Ⅳ期,但在位内膜ki-67表达在不同临床分期中无统计学差异(P>0.05)。结论:Fhit和ki-67可能与子宫内膜异位症有关。     

囊性纤维化跨膜转导调节因子在子宫内膜中表达的变化及其意义

目的　探讨囊性纤维化跨膜转导调节因子 (CFTR)在人月经周期子宫内膜中表达的变化及其意义。方法　采用免疫组化、免疫印迹、原位杂交方法检测人月经周期子宫内膜中CFTR的蛋白及mRNA的分布与含量变化。结果　 (1)CFTR仅在子宫内膜腺上皮细胞中表达 ,增生期晚期、分泌期、月经期内膜腺上皮细胞中均富含CFTR ,与增生期早期相比 ,差异有显著性 (P <0 0 5 ) ;免疫印迹法证实 ,增生期晚期、分泌期内膜存在特异性的、相对分子质量为 170 0 0 0的CFTR条带 ;(2 )CFTRmRNA在增生期晚期和分泌期早期子宫内膜中含量最高 ,与增生期早期、分泌期晚期相比 ,差异有显著性 (P<0 0 5 )。结论　人月经周期子宫内膜腺上皮细胞中存在CFTRmRNA和蛋白的表达 ,含量均随月经周期而变化 ;增生期晚期CFTR表达的增强。分泌期CFTR的表达及局部调节可能参与人类胚泡着床的调控。     

Wnt家族成员及其抑制子mRNA在人输卵管组织的表达

目的:证实Wnt家族成员及其抑制子在人输卵管的表达,并探讨孕激素及其拮抗剂对输卵管Wnt表达的调节作用。方法:选取生育年龄的正常妇女输卵管峡部组织,其中增生期、分泌期以及分泌期服用米非司酮50mg并于24h后手术的妇女输卵管组织各6例,应用实时荧光定量PCR(RT-PCR)技术检测输卵管组织的Wnt-3,Wnt-5a,Wnt-7a,Wnt-8b,DKK-1和FrpHEmRNA表达,用2-△△CT法分析结果。结果:Wnt-3,Wnt-5a,Wnt-7a,Wnt-8b,DKK-1和FrpHE的mRNA在人输卵管组织中均有表达。其中Wnt-5a,Wnt-7a,Wnt-8b在增生期与分泌期的表达无明显差异,Wnt-3和FrpHEmRNA在增生期表达明显高于分泌期(P<0.05),而DKK-1mRNA在分泌期的表达为增生期的1.6倍(P<0.05),服用米非司酮妇女的输卵管DKK-1表达明显下调(P<0.05)。结论:Wnt家族成员及其抑制子在人输卵管组织中存在表达,它们在月经周期不同时期的表达差异,可能是受到卵巢激素调控的结果。     

不同条件下小鼠围着床期子宫内膜整合素α_v和Le~y寡糖的表达

目的:探讨促排卵药、胚胎本身及孕后激素等条件对小鼠子宫内膜整合素av和Ley寡糖的表达及对着床窗开放的影响。方法:将成年雌鼠分为对照和实验两大组。对照组为自然受孕;实验组为用促排卵药后受孕,其中一部分为假孕,另一部分为事先行一侧输卵管结扎。各组分别于孕2-7 d取子宫标本2份,一份石蜡包埋行HE染色和免疫组化染色测定Ley寡糖,另一份于-80℃保存,冰冻切片行免疫组化染色测定整合素av。结果:①对照组于孕7 d,大部分小鼠子宫外观即见明显胚胎着床,显微镜下从孕5 d起出现子宫内膜间质的蜕膜反应,但实验组无1例有此变化。②整合素av在小鼠子宫内膜腺上皮细胞胞浆内表达。对照组的表达于孕4 d达高峰,而各实验组的表达高峰均推迟1-2 d,且表达水平较对照组明显减弱(P <0.05)。结扎侧和未结扎侧子宫内膜整合素av的表达高峰无明显差异。③Ley寡糖在小鼠着床期表达于子宫内膜腔上皮和腺上皮表面,其表达两组间无明显差异,不同妊娠天数之间的表达亦无明显差异。在对照组,于孕4 d开始Ley寡糖在子宫内膜间质有表达,孕6 d达高峰,而实验组未见一定的规律。结论:①小鼠子宫内膜腺上皮细胞胞浆内整合素av的表达与子宫内膜着床窗的开放一致。②促排卵药能抑制小鼠子宫内膜着床窗期整合素av的表达,延迟着床窗的开放;孕     

米非司酮治疗子宫内膜增生症前后ER和AR的表达及意义

目的:探讨米非司酮抑制子宫内膜增生的机制。方法:37例子宫内膜增生症(不伴非典型增生)患者,连续口服米非司酮(剂量10mg/d)治疗3个月,刮取治疗前、后子宫内膜组织,采用免疫组织化学二步法分别检测其ER和AR的表达进行自身对比;另取10例正常增殖期子宫内膜标本作为正常对照组进行比较。结果:子宫内膜增生症治疗前ER表达、AR表达均显著高于正常对照组(P<0.05),ER在腺体和间质细胞中均有表达,AR主要表达于间质细胞,腺体中几乎无表达。米非司酮治疗后腺体和间质ER表达比治疗前均下降(P<0.05),而间质和腺体中AR表达均比治疗前增加(P<0.05)。结论:米非司酮可通过下调ER和上调AR,从而抑制子宫内膜增生。     

基质金属蛋白酶-2、-9在子宫内膜异位症中的表达

目的 :探讨子宫内膜异位症患者在位及异位子宫内膜中基质金属蛋白酶 2、 9(MMP 2 ,MMP 9)的表达特点及在子宫内膜异位症发病机制中的作用。方法 :采用免疫组化SP法分别测定子宫内膜异位症 30例 (研究组 )、子宫肌瘤 2 1例 (对照组 )增生期和分泌期子宫内膜MMP 2、MMP 9的表达强度。结果 :对照组增生期和分泌期腺上皮细胞及基质细胞内MMP 2、MMP 9的表达呈阶段依赖性 ,分泌期高于增生期。在整个月经周期中 ,研究组在位及异位内膜中MMP 2、MMP 9的表达均持续高于对照组 (P <0 .0 5 )。结论 :MMP 2、MMP 9在子宫内膜的表达呈周期性变化 ,在位及异位内膜上MMP 2、MMP 9的持续过度表达可能与内异症的发生、发展有关     

子宫内膜增生性疾病患者内膜细胞凋亡的研究

目的 :研究凋亡在子宫内膜增生性疾病中的作用。方法 :用改良原位末端标记技术检测 15例正常月经周期的增生期、分泌期、月经期子宫内膜 ,11例增殖性子宫内膜 ,12例子宫内膜癌 ,以及术前用孕激素治疗的 13例异常增生子宫内膜中的凋亡细胞 ,并计算其凋亡指数 (AI)。结果 :分泌期、月经期子宫内膜、增殖性子宫内膜、子宫内膜癌AI均比正常增生期子宫内膜AI高 (P <0 .0 1)。增殖症患者内膜不典型增生组AI比单纯增生、复杂增生组AI高 (P <0 .0 5 ) ;内膜癌患者低分化组AI比高分化组、中分化组AI高 (P <0 .0 5 )。结论 :细胞凋亡与正常子宫内膜周期性变化有关 ,而在增殖性和癌变子宫内膜中的异常表达可能与子宫内膜的良恶性病变有关     

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     

Contraceptive use of antiprogestin.

OBJECTIVES: To study the effect of antiprogestin on ovarian function and endometrial development during the menstrual cycle and the possibility of using these compounds for contraceptive purposes. METHODS: Administration of different doses of the antiprogestin mifepristone during the menstrual cycle; intermittent measurements of luteinizing hormone, progestin and estrogen in blood and/or urine; endometrial morphology and concentration of markers for endometrial receptivity; efficacy trials of the contraceptive effect of mifepristone. RESULTS: A high dose of mifepristone administered in the follicular phase will inhibit follicular development. If mifepristone is given immediately after ovulation, the secretory development of the endometrium and the expression of, for instance, leukemia inhibitory factor and integrins will be inhibited. Similar effects on the endometrium are obtained with small weekly doses (2.5 or 5.0 mg) or small daily doses (0.5 mg) of mifepristone, which do not inhibit ovulation. Once-monthly administration of 200 mg mifepristone on the day after ovulation, and emergency postcoital treatment, are highly effective methods for preventing pregnancy. Even 5 mg once weekly has a significant contraceptive effect. CONCLUSIONS: The antiprogestin mifepristone has a number of effects during the menstrual cycle which makes the compound suitable for contraceptive use. Treatment after a single act of unprotected intercourse, and once-a-month treatment immediately after ovulation, have shown high contraceptive efficacy. A low-dose regimen which does not influence ovulation also has a contraceptive effect, but the efficacy needs to be improved before routine clinical use.     

长期应用小剂量米非司酮对子宫内膜形态和雌、孕激素受体的影响

本文利用HE染色和免疫组化方法结合计算机辅助图象分析技术观察了长期服用小剂量米非司酮对子宫内膜形态和雌、孕激素受体（ER、PR）的影响，15例子宫肌瘤患者于早卵泡期服用来非司酮（10例，10mg／日；5例，20mg／日）治疗，1～3个月后取内膜，另10例肌瘤患者取内膜作对照。结果：用药后所有的患者均闭经，内膜变薄，发育不良，无周期性改变，大部分基质致密，基质与腺体发育不同步，同时相间存在增殖与分泌期样改变，以增殖期样改变为主，腺体以分泌期样改变为主，腺体大小形态不一，但分泌不良。免疫组化显示内膜ER、PR明显分布不均，图象分析结果表明用药后内膜ER、PR相当于对照组增殖早中期水平。结果表明每日10mg米非司酮可明显抑制排卵并影响内膜成熟，在无孕激素存在的情况下，米非司酮有微弱的孕激素样作用。     

Expression of aminopeptidase N in human endometrium and regulation of its activity by estrogen

Objective: To determine whether aminopeptidase N (APN) regulates the cycle-dependent bioavailability of interleukin-8 (IL-8) in the endometrium.

Design: Prospective study.

Setting: University medical center.

Patient(s): Women without endometrial pathology from the proliferative (n = 25) or secretory (n = 18) phase of the menstrual cycle.

Intervention(s): We first immunolocalized APN in the endometrium using an anti-APN antibody. We then determined the regulation of APN kinetic activity by sex steroids in endometrial stromal cell cultures.

Main Outcome Measure(s): Expression of APN in human endometrium throughout the menstrual cycle. Regulation of APN activity by estradiol and progesterone in cultured endometrial stromal cells.

Result(s): Immunohistochemistry of endometrial sections revealed staining of endometrial stroma throughout the menstrual cycle. There was no detectable staining in glandular cells. The expression of APN as detected by immunohistochemistry was significantly lower in the early proliferative phase. In cultured cells, estradiol inhibited APN activity in a concentration-dependent manner. Progesterone did not have a significant effect.

Conclusion(s): Stromal localization of APN in endometrium may explain the epithelial rather than stromal presence of IL-8 in vivo. Decreased expression of APN may increase IL-8 bioavailability thus contributing to angiogenesis and polymorphonuclear leukocyte chemotaxis in early proliferative phase.     

Expression of complement system regulatory molecules in the endometrium of normal ovulatory and hyperstimulated women correlate with menstrual cycle phase

The present study describes the temporal expression of complement regulatory molecules membrane cofactor protein (MCP), decay accelerating factor (DAF), CR1, and CD59 in the human endometrium throughout the normal menstrual cycle and in patients submitted to ovarian hyperstimulation. During its proliferative phase, the endometrium expresses MCP, with increased expression during the secretory phase. Phase-dependent expression also was observed for DAF and CD59, mainly in the secretory phase. Expression of CR1 was not detected. These results suggest the presence of complement system activity during the menstrual cycle, with greater expression of regulatory molecules during the secretory phase to protect the epithelial integrity of human endometrium.     

Effect of mifepristone on estrogen and progesterone receptors in human endometrial and endometriotic cells in vitro

OBJECTIVE: To investigate the expressions of estrogen (E) receptor and progesterone (P) receptor in human eutopic and ectopic endometrium and the effect of mifepristone (RU486) on them. DESIGN: Prospective study. SETTING: University hospital. PATIENT(S): Twenty-two patients with ovarian endometriosis and 13 patients with uterine leiomyoma were recruited. INTERVENTION(S): Samples of ovarian endometrioma cyst tissue and endometrium were obtained from the 22 patients. A sample of endometrium was obtained from the 13 patients. MAIN OUTCOME MEASURE(S): Expressions of E and P receptors were determined using immunocytochemical method. RESULT(S): P receptor expression in endometrial epithelial cells with endometriosis was significantly higher than that without endometriosis in the early secretory phase. Estrogen receptor and epithelial P receptor expressions in endometriotic cells were significantly lower than those of endometrial cells during the proliferative phase, similar with the latter during the early secretory phase and significantly higher during the late secretory phase. RU486 down-regulated the expressions of E and P receptors in both the eutopic and the ectopic endometrial cells, and in some cases this down-regulating effect was more apparent when the concentration of RU486 was higher. CONCLUSION(S): Different steroid receptor expressions indicate different hormonal regulation between endometriotic and endometrial cells. The down-regulating effect on E and P receptors may be one of the therapeutic mechanisms of RU486 on endometriosis.     

Hoxa-10在人月经周期分泌中期子宫内膜腺上皮表达下降

目的 :对同源异型框基因 Hoxa-1 0在月经周期各阶段人子宫内膜的表达形式进行观察。方法 :通过原位杂交方法测定了 5例增殖期、9例分泌期的人子宫内膜 Hoxa-1 0的表达。结果 :Hoxa-1 0基因在人子宫内膜均有表达。但在各周期阶段 ,其表达的强度和部位却有所不同。首次发现 Hoxa-1 0在对应于着床期的分泌中期以及分泌晚期子宫内膜腺上皮的表达明显降低或不表达。结论 :Hoxa-1 0在人类着床过程中可能具有重要功能。     

Metallothionein and RCAS1 expression in comparison to immunological cells activity in endometriosis, endometrial adenocarcinoma and endometrium according to menstrual cycle changes

OBJECTIVE: Endometrium is a specialized organ in which phenomena controlling the level of cell proliferation and apoptosis are marked. The aim of our study was to determine the presence of proteins involved in apoptosis and proliferation: RCAS1, MT and the number of CD56-positive cells and their activity to elucidate their possible role in the development of adenocarcinoma and endometriosis. MATERIALS AND METHODS: MT, RCAS1, CD56-positivity and CD69 expression were assessed in 55 tissue samples by Western blot and immunohistochemistry methods. RESULTS: We found that endometrium during secretory menstrual cycle phase is characterized by significantly higher RCAS1 and higher MT expression than in proliferative phase. The number of CD56-positive cells and the CD69 antigen expression was significantly increased. Endometrial adenocarcinoma was characterized by significantly increased RCAS1 expression, while MT expression was comparable to the level found in the secretory phase. The number of CD56-positive cells was significantly decreased and their activity was comparable to the level found in the secretory phase. Endometriosis was accompanied by significantly lower RCAS1 and MT expressions, with lower number of CD-56 positive cells and lower expression of CD69 antigen in comparison to the secretory phase. CONCLUSIONS: The ability of endometrium to determine cytotoxic activity (RCAS1 expression changes) and high protection against DNA damage (MT expression) with concomitant changes in the number of immune cells and their activity, observed in normal endometrium during the menstrual cycle phases seems to be fundamental for pathological features of endometrial adenocarcinoma and endometriosis.