Norplant皮下埋植后子宫内膜形态和雌、孕激素受体含量的变化

目的探讨Ｎｏｒｐｌａｎｔ皮下埋植后子宫内膜形态和雌、孕激素受体（ＥＲ、ＰＲ）含量的变化与子宫异常出血的关系。方法应用免疫组化、ＷｅｓｔｅｒｎＢｌｏｔ和原位杂交技术结合计算机图像分析,观察１６例Ｎｏｒｐｌａｎｔ皮下埋植后和２３例正常周期的内膜的形态学改变、细胞增殖状态及ＥＲ、ＰＲ蛋白及其ｍＲＮＡ的表达。结果埋植Ｎｏｒｐｌａｎｔ后,内膜ＤＮＡ含量降低;螺旋动脉数量减少,腺体的构成比例降低;腺体构成比例与雌二醇（Ｅ２）水平呈正相关,与子宫异常出血呈负相关,与年龄和体重无相关性,随使用期延长,腺体构成比增加。埋植后,内膜ＥＲ含量低于月经周期各期水平,但ＰＲ含量相当于或高于月经周期最高水平。ＥＲ、ＰＲｍＲＮＡ表达减弱,以ＥＲｍＲＮＡ表达降低更明显。结论内膜腺体和螺旋动脉再生修复不良和ＥＲ、ＰＲ表达异常,可能是Ｎｏｒｐｌａｎｔ致子宫异常出血的病理学基础     

Effects of estradiol on in vitro maturation of buffalo and goat oocytes

PurposeThe effects of estradiol on oocyte development seem to be varied among species. The present study investigated the effects of 17β‐estradiol on in vitro maturation of buffalo and goat oocytes.MethodsCumulus oocyte complexes (COCs) were aspirated from large antral follicles of slaughtered buffalo and goat ovaries. COCs were cultured in TCM‐199 medium supplemented with 0, 0.5, 1, and 1.5 µg/mL of 17β‐estradiol for in vitro maturation. Then, oocytes were used for the examination of state of nuclear maturation and cumulus expansion.ResultsIn both species, oocytes treated with 17β‐estradiol showed higher cumulus expansion rate than control (0 µg/mL treated). In buffalo, the percentage of oocytes matured to the metaphase II (MII) stage increased in the concentration‐dependent manner of 17β‐estradiol. Similarly, estradiol positively influenced nuclear maturation of goat oocytes in vitro.ConclusionsEstradiol has promoting effects on normalprogress of in vitro oocyte meiosis in buffalos and goats.     

乳腺癌激素受体的表达与新辅助化疗疗效的相关性

目的 研究乳腺癌雌孕激素受体的表达与新辅助化疗疗效之间的相关性。方法 用免疫组化的方法检测46例乳腺癌患者在接受新辅助化疗之前雌孕激素受体的表达情况，新辅助化疗结束后进行疗效评价。结果 46例患者中，临床完全缓解(cCR)17例，部分缓解(PR)21例，无变化(NC)8例，其中病理完全缓解(pCR)9例，总有效率为82．6％，pCR率为19．57％。在受体双阴的22例患者中，20例有效(cCR PR)，总有效率为90．9％，NC2例；pCR6例(27．3％)，受体单阳或双阳的2，4例患者中，18例有效(cCR PR)，总有效率为75．0％，NC6例。pCR3例(12．5％)。受体双阴组患者的pCR率及总有效率都高于受体单阳或双阳组患者，但未见显著性差异。结论 受体双阴的患者可能较受体单阳或双阳的患者对化疗的敏感性更高。     

子宫肉瘤雌、孕激素受体表达与临床生物学特性及预后的关系

目的　研究子宫肉瘤雌、孕激素受体的表达情况以及其临床生物学意义。方法　选取 1 991年9月～ 2 0 0 1年 9月北京大学人民医院及第三医院收治的子宫肉瘤患者 4 4例 ,选取石蜡病理切片 ,用免疫组化法测定ER、PR表达情况。结果　①ER表达阳性率在子宫内膜间质肉瘤中为 75 % ,子宫平滑肌肉瘤为 31 8% ,子宫恶性苗勒管混合瘤为 2 /6。子宫内膜间质肉瘤ER表达阳性率显著高于子宫平滑肌肉瘤 (P <0 0 5 ) ;②PR表达阳性率在子宫内膜间质肉瘤中为 1 0 0 % ,子宫平滑肌肉瘤为 6 8 2 % ,子宫恶性苗勒管混合瘤为 3/6。子宫内膜间质肉瘤的PR表达阳性率显著高于子宫平滑肌肉瘤及恶性苗勒管混合瘤 (P <0 0 5 ) ;③与临床病理学特征的关系 :ER与PR间呈显著正相关性。无瘤存活者ER、PR表达率显著高于因瘤死亡者 (P <0 0 5 )。核分裂 <1 0个 /1 0HPs的子宫肉瘤PR表达阳性率显著高于核分裂≥ 1 0个 /1 0HPs的子宫肉瘤 (P <0 0 5 )。结论　①子宫内膜间质肉瘤ER、PR阳性率均显著高于子宫平滑肌肉瘤和子宫恶性苗勒管混合瘤 ,子宫内膜间质肉瘤患者可常规进行ER、PR测定 ,根据ER及PR表达情况辅助激素治疗 ;②无瘤存活者ER、PR表达率显著高于因瘤死亡者 ,说明ER、PR表达与子宫肉瘤的预后有关     

孕三烯酮对大鼠血清雌孕激素及在位和异位子宫内膜雌孕激素受体的影响

目的:探讨孕三烯酮(gestrinone)治疗子宫内膜异位症的作用机理。方法:用自体移植术建立大鼠子宫内膜异位症动物模型,随机分组给药,3周后剖腹,取血测定雌二醇(E2)、孕酮(P)浓度,取出子宫及移植物,固定、切片,免疫组化SP法测定各组动物子宫及移植物的雌激素受体(ER)和孕激素受体(PR)的表达。结果:孕三烯酮高剂量使大鼠血清E2、P水平降低,而中低剂量对E2、P水平影响不大。免疫组化结果则显示移植物的ER、PR含量较子宫内膜低,孕三烯酮高、中剂量可下调在位和异位子宫内膜ER、PR表达,而低剂量孕三烯酮则不能明显改变ER、PR的表达量。结论:孕三烯酮可通过降低体内雌孕激素水平以及下调在位及异位子宫内膜ER、PR的表达,发挥对子宫内膜异位症的治疗作用。     

Evaluation of 14-3-3 protein family levels and associated receptor expression of estrogen and progesterone in Human Uterine Leiomyomas

Objective: Uterine leiomyomas represents a major public health problem. Despite their prevalence, the causation and pathogenesis of leiomyomas are poorly understood. A broad range of organisms and tissues contain 14-3-3 proteins which have been associated with the pathogenesis of many diseases through participating in signal transduction pathways. This study was designed to evaluate which 14-3-3 isoforms might be optimal targets in leiomyomas, and to further explore their relationship with estrogen and progesterone receptor (ER and PR). Methods: Paired samples of leiomyoma and adjacent myometrium were obtained from 80 subjects who had surgical excision of uterine leiomyomas. The expression of 14-3-3 isoforms was detected by Western bolt and RT-PCR, and their relationship with ER and PR was analysed by immunohistochemistry. Results: The expressions of 14-3-3σ had decreased significantly in leiomyoma compared with that in normal myometrium and was negatively correlated with ER and PR by immunohistochemistry. Conclusion: The down-regulation of 14-3-3σ in leiomyoma suggests that 14-3-3σ may play a role in tumorigenesis, and that its mechanism may be involved in the up-regulation of ER and PR.     

17β雌二醇对人精子功能的非基因组调节效应及其机制的研究

目的：探讨17β雌二醇对人精子功能非基因组效应的可能机制。方法：运用计算机辅助精子分析系统与人精子穿透去透明带金黄地鼠卵异种体外受精实验,评价17β雌二醇对精子功能的调节作用；精子经信号转导途径抑制剂处理后,再采用流式细胞术检测精子胞内Ca~(2+)浓度的变化。结果：1μmol/L和5μmol/L E_2-BSA能提高人精子的A级精子百分比、平均曲线运动速度、平均直线运动速度、平均路径速度(P＜0．05)；自身受精率偏低的继发不育患者精子经1μmol/L和5μmol/L E_2-BSA作用后受精率提高(P＜0．05),但对本身受精率较高的精子影响不大(P＞0．05)；应用腺苷酸环化酶、磷脂酶C和酪氨酸蛋白激酶特异性抑制剂分别作用后均明显抑制E_2-BSA引起的胞内Ca~(2+)浓度的上升(P＜0．05)。结论：17β雌二醇能显著提高精子的运动能力和受精率,17β雌二醇可能通过腺苷酸环化酶、磷脂酶C或酪氨酸蛋白激酶信号转导途径实现对人精子功能调节的非基因组效应,为改善人精子功能提供了可能的新途径。     

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.     

小鼠子宫内膜上皮细胞表面肝素受体及性甾体激素对它的影响

取5～6周龄ICR 小鼠子宫,用酶消化,分离出上皮细胞,并接种于24孔培养碟中,待细胞长成片后,加入不同剂量[~3H]标记的肝素孵化。实验结果表明,上皮细胞表面肝素受体量,随[~3H]肝素含量增加而增加,直至饱和为止。另外,上皮细胞上肝素受体量也与[~3H]标记肝素孵化时间相关。随着孵化时间增加,其表面受体含量也增加,30min 基本饱和。在培养液内加入0.045μmol/L 及0.09μmol/L 雌二醇,其结果表明,雌二醇可增加小鼠子宫内膜上皮细胞表面肝素受体的含量,但两种剂量组间未见明显差异。     

雌激素对去势小鼠脾细胞凋亡的作用机制探讨

目的:探讨雌激素对去势小鼠脾细胞凋亡、凋亡基因Fas、FasL及ER亚型表达的影响。方法:流式细胞仪检测不同浓度E2时去势小鼠脾细胞的凋亡率;RT-PCR方法检测细胞内Fas、FasL及ERα和ERβmRNA的表达。结果:除最低浓度(10-5μmol/L)外,去势+E2组凋亡率及Fas、FasL基因的表达都高于青年组(P<0.05);与去势对照组相比,E2在10-3μmol/L、10-2μmol/L浓度时凋亡率及Fas、FasL基因的表达增高(均P<0.01)。E2在10-4μmol/L时ERα、ERβ基因的表达升高(ERα:P<0.05;ERβ:P<0.01),以后随浓度增加ERα和ERβmRNA水平有下降去势。结论:高浓度E2致脾细胞凋亡,与上调凋亡促进基因Fas、FasL基因表达有关,低浓度E2可能逆转去势小鼠脾细胞凋亡。不同浓度E2对去势小鼠脾细胞ERα和ERβ基因表达的影响支持雌激素对ER表达的调节作用:在生理范围内为正向调节,超生理范围为负向调节。     

Ovarian cancer in infertile women during or after ovulation-inductiontherapy: expression of LH/hCG receptors and sex steroid receptors

Kuroda H, Konishi I, Mandai M, Nanbu K, Rao Ch V, Mori T. Ovarian cancer in infertile women during or after ovulation-induction therapy: expression of LH/hCG receptors and sex steroid receptors. Int J Gynecol Cancer 1997; 7: 451–457.
A possible association between the use of fertility drugs and development of ovarian cancer has recently been suggested. Review of 131 patients with ovarian carcinoma in our hospital revealed that six (4.6%) cases had developed during or after ovulation-induction therapy. To examine the hormone sensitivity of these tumors, the expression of luteinizing hormone/chorionic gonadotropin (LH/hCG) receptors and sex steroid receptors in the tumor cells was analyzed by immunohistochemistry. In addition, to elucidate the clinical and pathological characteristics of ovarian cancer in this setting, all of the 41 cases, including both of our series and previously reported cases in the literature, were reviewed. Immunohistochemical analysis showed that expression of LH/hCG receptors or sex steroid receptors in the tumor cells was observed in five of the six cases. This suggests that gonadotropins and/or sex steroids may influence the biological behavior of the tumor. Ovarian cancer during or after ovulation-induction consisted of two different types: one is serous, low-malignant potential (LMP) or invasive carcinoma and the other is endometrioid or clear cell carcinoma arising in an endometriotic cyst. Although the latter group is usually detected at an early stage, serous tumors are frequently at an advanced stage, even under close examination during infertility treatment. This is essential for giving informed consent during ovulation-induction therapy for infertile women.     

Breast cancer and sex steroids: Critical review of epidemiological,experimental and clinical investigations on etiopathogenesis,chemoprevention and endocrine treatment of breast cancer

There is strong epidemiological, experimental and clinical evidence that the etiology of breast cancer is closely related to long-term exposure of breast epithelium to sex steroid hormones. Estrogens can enhance the development of breast cancer by stimulating cell proliferation rate and thereby increasing the number of errors occurring during DNA replication, as well as by causing DNAdamage via their genotoxic metabolites produced during oxidation reactions. Anti-estrogenic drugs, including tamoxifen, raloxifene and anastrozole, have been tested with promising results in the chemoprevention of breast cancer in high-risk women. As for the use of exogenous sex-steroids in the gynecological practice, data about breast cancer risk associated with oral contraception are reassuring, and available data on oral hormone replacement therapy (HRT) use for not more than 5 years have failed to detect a significant increase in the risk of developing a breast cancer. Long-term HRT administration increases the incidence of this tumor slightly, with a relative risk ranging from 1 to 2 depending on hormone preparation. Estrogens alone, even if taken for long periods of time, seem to be safer than estrogen/progestin combinations. New administration routes and novel hormone regimens are currently under evaluation,.and these new HRT modalities could have different impact on breast cancer risk because of their metabolic and pharmacodynamic effects.     

Effects of estradiol and progestogens on tumor-necrosis factor-alpha-induced changes of biochemical markers for breast cancer growth and metastasis

Background. Epidemiological data suggest an enhanced breast cancer risk during estrogen/progestogen therapy as compared to estrogen monotherapy in postmenopausal women. The underlying mechanism, however, still remains unknown. Estrogens are known to be mitogenic agents for benign and cancerous breast epithelial cells whereas the role of progestogens is unclear. Tumor-associated macrophages play a crucial role in tumor growth and metastasis due to the synthesis of various cytokines such as tumor necrosis factor-α (TNF-α), which can stimulate the synthesis of proliferative and angiogenic factors in tumor cells. In an in vitro model we investigated the influence of estradiol and estradiol/progestogens combinations on the changes of TNF-α- induced markers.

Methods. MCF-7 cells, a human estrogen- and progesterone-receptor-positive human breast cancer cell line, were used for the experiments. Estradiol (E₂), at a concentration of 0.1 nM, and the progestogens progesterone (P), norethisterone (NET) and medroxyprogesterone acetate (MPA), each at concentrations of 0.01 to 1 μM, were tested alone and in combination. The cells were incubated for 4 days and the markers monocyte chemoattractant protein-1 (MCP-1), vascular endothelial growth factor (VEGF) and matrix metalloproteinase-9 (MMP-9) were measured in the supernatant by enzyme-linked immunosorbent assay.

Results. E₂ in combination with TNF-α elicited significant increases in MCP-1 and VEGF concentrations compared with TNF-α alone. For the progestogens alone an increase of MCP-1 was observed for NET, whereas MPA induced a decrease. An increase of VEGF was observed for all progestogens, the effect being greatest for MPA. No changes were found for MMP-9. In combinations with E₂, the E₂-induced increase of MCP-1 was reduced by NET and MPA and the increase of VEGF was diminished by P and NET, but not by MPA. The E₂-induced decrease of MMP-9 was not antagonized by P and NET, but completely abolished by MPA.

Conclusion. Our results indicate that E₂ may have a stimulating effect on pre-existing tumor growth and metastasis. This effect seems to be influenced by progestogens in a different manner. Thus the choice of progestogen addition to estrogen therapy may be important, especially since different effects can occur in the case of pre-existing tumor cells.     

Association of endometriosis risk and genetic polymorphisms involving biosynthesis of sex steroids and their receptors: an updating meta-analysis

The objective of our study is to assess the association of endometriosis risk and genetic polymorphisms involving biosynthesis of sex steroids and their receptors. A systematic search of three databases was conducted. Twenty-seven studies on the association of the cytochrome P450 subfamily 17 (CYP17), estrogen receptor gene (ER), progesterone receptor gene (PR), 17-beta-hydroxysteroid dehydrogenase type 1 gene (HSD17B1), and cytochrome P450 subfamily 19 (CYP19) polymorphisms with endometriosis risk were identified. When all groups were pooled, we found an association between HSD17B1 (A variant allele vs. G wild allele: odds ratio (OR)=1.42, 95% confidence interval (CI)=1.10-1.84, P=0.007) and PR (P2 variant allele vs. P1 wild allele, OR=1.43, 95% CI=0.99-2.08, P=0.058) polymorphisms and endometriosis risk, while failing to detect links with CYP17, ER, and CYP19 polymorphisms examined. In the subgroup analysis, a significant association of CYP17 and ERα-PvuII polymorphisms with endometriosis was found neither in a Caucasian population nor in an Asian population. The findings of our study suggest that HSD17B1 and PR polymorphisms are associated with an increased risk of endometriosis. Further investigation into the association between CYP17, ER, PR, HSD17B1, and CYP19 polymorphisms and endometriosis risk is warranted and should include larger sample sizes.     

Effects of phytoestrogens genistein and daidzein on progesterone and estrogen (estradiol) production of human term trophoblast cells in vitro

Objective.?Phytoestrogens are a diverse group of nonsteroidal plant compounds that occur naturally in many plants. Because they possess a ring system similar to estrogens they are able to bind on estrogen receptors alpha and beta in humans. The effects of the phytoestrogens genistein and daidzein on the production of progesterone and estrogen in isolated human term trophoblast cells in vitro were tested in this study.

Material and methods.?Cytotrophoblast cells were isolated from human term placentas. Phytoestrogens genistein and daidzein were incubated in different concentrations with trophoblast cells. Untreated cells were used as controls. After 24 h aliquots were removed and tested for progesterone and estrogen production.

Results.?The production of the steroid hormones progesterone and estrogen are influenced by phytoestrogens genistein and daidzein in human term trophoblast cells. A strong inhibition effect of both phytoestrogens tested in the production of progesterone was demonstrated. In addition, a significant stimulating effect on estrogen production by genistein and daidzein was observed.

Conclusion.?Results obtained with this study show that phytoestrogens (genistein and daidzein) sufficiently reduce progesterone production in human term trophoblast cells. Because blockade of progesterone is a possible mechanism involved in initiation of labor, we may speculate that high doses of phytoestrogens at the feto-maternal interphase could play a negative role in maintenance of pregnancy. Stimulation of estrogen production by genistein and daidzein in trophoblast cells is probably due to estrogen receptor blocking effects of both phytoestrogens. Trophoblast cells seem to compensate blocking of its estrogen receptors by higher estrogen production.     

Effects of tibolone and its metabolites on prolactin and insulin-like growth factor binding protein-1 expression in human endometrial stromal cells

Abstract

The effects of the postmenopausal replacement steroid tibolone and its 3α-, 3β-OH and Δ-4 tibolone metabolites were evaluated on progesterone receptor-mediated classic decidualization markers insulin-like growth factor binding protein-1 (IGFBP-1) and prolactin expression in human endometrial stromal cells (HESCs). Supernatants of conditioned medium or erxtracted RNA from experimental cell incubations of confluent HESCs were subjected to ELISAs, Western blot analysis and RT/PCR, and results were statisically assesed. Over 21 days, specific ELISAs observed linear increases in secreted IGFBP-1 and prolactin levels elicited by tibolone and its metabolites. Cultured HESCs were refractory to E2 and dexamethasone, whereas tibolone and each metabolite exceeded medroxyprogesterone acetate in significantly elevating IGFBP-1 and prolactin output. Anti-progestins eliminated IGFBP-1 and prolactin induction by tibolone and its metabolites. Immunoblotting and RT/PCR confirmed ELISA results. These observations of IGFBP-1 and prolactin expression: (a) indicate the relevance of cultured HESCs in evaluating the chronic effects of tibolone administration to women; (b) are consistent with PR-mediated endometrial atrophy and protection against endometrial bleeding despite the persistence of circulating ER-binding, but not PR-binding metabolites following tibolone administration to women.     

RU_(486)对早孕妇女蜕膜组织甾体激素受体及内分泌的影响

目的 了解米非司酮及米索对人早孕蜕膜雌激素受体(ER)、孕激素受体(PR)的表达及血清内分泌水平的影响。方法 取正常早孕、服用米非司酮及服米非司酮配伍米索后各20例的蜕膜,应用单克隆抗体链霉素亲生物蛋白——过氧化酶 (SP) 免疫组织化学方法测定 ER、PR 的表达及血清激素水平的变化。结果米非司酮配伍米索使蜕膜组织中 ER、PR 表达下调,正常早孕组、服米非司酮组及服米非司酮配伍米索组之间差异有显著性。血清雌二醇(E_2)、睾酮(T)、泌乳素(PRL)含量上升,孕酮(P)下降;而对 FSH、hCG 的含量无明显影响。结论 大剂量RU_(486)能竞争地结合蜕膜组织孕激素受体,使早孕期高浓度的内源性孕酮与其受体的正常结合受到干扰,从而达到抗早孕的目的。