雌激素及植物雌激素对去卵巢大鼠子宫组织VEGF表达和形态的影响

目的比较雌激素及植物雌激素对去卵巢大鼠子宫组织VEGF和形态表达的影响。方法48只Wistar大鼠随机分为8组:假手术组(Sham)、去卵巢组(Ovx)、17β-雌二醇组(Ovx+Est)、17-β雌二醇+黄体酮组(Ovx+Est+Pro)、黄体酮组(Ovx+Pro)、染料木素组(Ovx+Gen)、白黎芦醇组(Ovx+Res)、根皮素组(Ovx+Phl)。给药21 d,用蛋白免疫印迹法观察各组子宫VEGF的表达、并用免疫组化对VEGF的表达部位和其形态学变化进行观察。结果与Sham组比较,Ovx组的VEGF表达下调;与Ovx比较,Ovx+Est和Ovx+Est+Pro两组VEGF表达均明显上调。Gen、Res和Phl对卵巢切除大鼠VEGF无影响。大鼠卵巢切除后,子宫明显萎缩且重量下降。皮下注射Est或Est合并Pro后,子宫组织密度增加,子宫被覆上皮细胞和腺体明显增生。皮下注射Gen和Phl后,子宫形态变化类似卵巢切除大鼠,注射Res可促进去卵巢大鼠子宫腺体的增生,但作用没有雌激素强。结论雌孕激素使VEGF表达明显上调;植物雌激素Res有促进腺泡增生作用。     

雌激素替代治疗对去势大鼠阴道组织雌激素受体表达的影响

目的 研究雌激素对去势大鼠阴道组织ERα与ERβ表达的影响，探讨雌激素治疗老年性阴道炎的分子机制。方法将40只雌性SD大鼠随机分成对照组、假手术组、去势组和雌激素替代治疗组各10只。对照组为空白对照；假手术组去除卵巢周围与卵巢相同大小脂肪组织；去势组行双侧卵巢切除术；雌激素替代治疗组在去卵巢手术3周后行戊酸雌二醇800 μg•kg 1•d 1替代治疗8周。免疫组织化学染色检测大鼠阴道组织ERα与ERβ表达情况。结果去势组大鼠ERα、ERβ表达显著低于对照组（均P＜0.01），雌激素替代组ERα、ERβ表达上升，较去势组显著增强（均P＜0.01）。结论雌激素替代治疗可导致雌激素受体表达上升，可能导致雌激素受体效应增强，从而提高阴道局部抵抗力。     

雌激素抑制大鼠肝癌肺转移机制研究

目的 探讨雌激素对大鼠肝癌肺转移形成的影响及其可能的机制.方法 将雌性大鼠随机分为切除卵巢组、对照组、假手术组和去卵巢后加雌激素组,建立大鼠肝癌模型.诱癌20周后观察肿瘤发生、肺转移的情况;免疫组织化学法检测4组成瘤大鼠肝癌组织中雌激素受体α(ERα)、磷酸化雌激素受体α(P-ERα)、肝细胞生长因子(HGF)和白介素6(IL-6)的表达;半定量逆转录聚合酶链反应法检测ERα、HGF及IL-6的mRNA水平.结果 切除卵巢组大鼠肝癌肺转移发生率明显高于其他3组(P<0.05);大鼠肝癌肺转移发生率越高,肝癌组织中HGF、IL-6的蛋白及分子水平表达越高(P<0.05),而ERα的蛋白及分子水平表达越低(P<0.05).结论 雌激素可能通过下调HGF和IL-6的表达来抑制肝癌肺转移的发生.     

乳结平胶囊对乳腺增生大鼠乳腺组织ER、PR表达的影响

目的:探讨乳结平胶囊对乳腺增生大鼠病理组织雌激素受体(ER)和孕酮受体(PR)表达的影响。方法:先采用苯甲酸雌二醇注射液连续im25d,后改用黄体酮注射液连续im5d,从而建立大鼠乳腺增生模型,以三苯氧胺为阳性对照药物,用乳结平胶囊高、低剂量治疗30d。结果:三苯氧胺组及乳结平胶囊高、低剂量组均能减轻大鼠乳头红肿或增生症状,降低大鼠血清雌二醇(E2)、催乳素(PRL)水平及升高孕酮(P)水平,降低子宫指数,明显减少导管上皮细胞层数和腺泡数,同时减少乳腺组织ER、PR的含量,抑制乳腺组织增生。结论:乳结平胶囊可通过调节雌激素水平,改善增生乳腺的病理形态变化,从而有效治疗大鼠乳腺增生。     

两种雌激素对去卵巢大鼠子宫内膜的影响

目的：探讨戊酸雌二醇和炔雌醇对去卵巢大鼠子宫内膜的作用,为绝经后妇女雌激素补充治疗提供理论依据。方法选用3月龄雌性Sprague-Dawley（ SD）大鼠40只,实验分为Sham组（假手术组）、Ovx组（去卵巢组）、A组（炔雌醇组）、B组（戊酸雌二醇组）。除Sham组外,其余各组均切除双侧卵巢建立骨质疏松动物模型。去势后4周开始A组炔雌醇（0.1 mg·kg-1·d-1）、B组戊酸雌二醇（0.4 mg·kg-1·d-1）灌胃。4组均在90 d对大鼠称重,取大鼠子宫进行称重,子宫内膜行病理切片检查。结果用药12周后各组间大鼠体重无统计学差异。 B 组子宫湿重及子宫指数明显低于A组（P〈0.05）,与Ovx组比较,无显著性差异（P〉0.05）。但A组子宫湿重及子宫指数明显升高,与Ovx组比较差异有统计学意义（P〈0.05）。两组用药组与Ovx组相比均显示子宫内膜增厚,上层单层柱状,肌层肌纤维排列规则一致,浆膜层光滑。 B组可见内膜增厚,但低于A组,两组均未见内膜增生过长、核异性和非典型增生样改变。结论戊酸雌二醇与炔雌醇用于绝经后雌激素补充治疗,虽对子宫内膜均有生长作用,但均未见过度生长及异型性改变。     

中药益坤宁对围绝经期大鼠卵巢雌激素受体表达的影响

目的观察中药益坤宁对围绝经期大鼠卵巢雌激素受体(estrogen receptor,ER)α、ERβmRNA和蛋白表达的影响。方法12月龄雌性Wistar大鼠随机分为老年模型组、西药利维爱组、中药益坤宁组,连续灌胃给药4周;以5月龄大鼠作为青年对照组。采用RT-PCR和免疫组织化学检测大鼠卵巢中ERα、ERβmRNA和蛋白的表达。结果RT-PCR和免疫组织化学结果都显示:老年模型组大鼠卵巢ERα、ERβmRNA和蛋白表达明显低于青年组(P<0.05),中药益坤宁组和西药利维爱组ERα、ERβmRNA和蛋白表达明显升高,与老年模型组相比有显著性差异(P<0.05或P<0.01)。结论中药益坤宁上调大鼠卵巢ERα、ERβ的表达,可能是治疗围绝经期综合征的作用机制之一。     

雌激素预处理增强肝缺血再灌注损伤大鼠雌激素受体表达

目的探讨雌激素预处理对肝缺血再灌注(I/R)损伤大鼠雌激素受体(ERα)表达的影响。方法采用Pringle法阻断大鼠第一肝门制作95%肝I/R模型。肝缺血前1 h大鼠ip给予雌二醇4 mg·kg-1(雌二醇+I/R组)。分别取肝缺血前(0)、再灌注1,3,6和24 h共5个时间点的血液标本,检测血清谷丙转氨酶(ALT)水平;ELISA法测定血清肿瘤坏死因子-α(TNF-α)水平;HE染色观察肝组织形态学变化,免疫组化方法检测肝核因子κB(NF-κB)和ERα的表达。结果与假手术组相比,I/R模型组和雌二醇+I/R组血清ALT和TNF-α水平、肝组织NF-κB和ERα表达明显升高(P<0.01);与假手术组相比,再灌注6 h,I/R和雌二醇+I/R组血清ALT水平分别升高了11.7倍和8.1倍(P<0.01),TNF-α水平分别升高5.2倍和3.2倍(P<0.01),但雌二醇+I/R组升高幅度明显低于I/R组(P<0.05);与假手术组相比,再灌注6 h I/R组和雌二醇+I/R组肝组织NF-κB表达分别升高了8.6倍和4.8倍,但雌二醇+I/R组升高幅度明显低于I/R组(P<0.01);ERα表达分别升高了4.2倍和7.4倍,雌二醇+I/R组升高幅度明显高于I/R组(P<0.01);雌二醇+I/R组肝组织损伤明显轻于I/R组。结论雌激素预处理可减轻I/R导致的肝组织损伤程度,此作用与增强ERα表达有关。     

当归芍药散对围绝经期模型大鼠子宫结构及雌激素受体表达的影响

目的:探讨当归芍药散(DSS)对围绝经期模型大鼠子宫结构及子宫腔上皮和基质中雌激素受体α(ERα)、雌激素受体β(ERβ)表达的影响。方法:将40只雌性SD大鼠随机分为假手术组(生理盐水)、模型组(生理盐水)和DSS低、中、高剂量组(1.94、3.87、7.44 g/kg),每组8只。除假手术组大鼠切除卵巢附近脂肪外,其余各组大鼠切除双侧卵巢以建立围绝经期模型。造模成功后,大鼠每天ig给药1次,连续8周。给药结束后,称定大鼠子宫湿质量,观察大鼠子宫形态和结构的变化,测定子宫腔上皮及基质中ERα、ERβ表达水平。结果:与假手术组比较,模型组大鼠子宫内膜柱状上皮呈低柱状,固有层、肌层与浆膜层均显著萎缩,基质细胞可见明显核固缩,子宫湿质量、宫腔面积、内膜厚度及腺体数量均显著减少(P<0.01),子宫腔上皮及基质中ERα、ERβ表达水平均显著降低(P<0.01)。与模型组比较,DSS各剂量组大鼠子宫内膜及固有层萎缩程度差异无统计学意义,但固有层内腺体丰富,子宫湿质量、宫腔面积、内膜厚度以及子宫腔上皮和基质中ERα表达水平差异均无统计学意义(P>0.05),但DSS中、高剂量组大鼠子宫腺体数量显著增加(P<0.01),DSS高剂量组大鼠子宫腔上皮及基质中ERβ表达水平显著升高(P<0.05或P<0.01)。结论:DSS对围绝经期模型大鼠子宫腺体萎缩症状的改善作用不明显,但可增加模型大鼠的腺体数量,这可能与提高子宫腔上皮及基质中ERβ表达水平有关。     

雌激素对卵巢切除后雌性大鼠心肌组织中雌激素受体表达的影响

目的：探讨雌激素对卵巢切除后雌性大鼠心肌组织中雌激素受体表达的影响。方法：雌性SD大鼠卵巢切除术（OVX）后1周,随机分成3组：OVX＋雌激素（0.15mg/kg,s.c.）、OVX＋生理盐水、对照组。4周后,Westernblotting检测α和β两种雌激素受体在心肌组织中的表达。结果：卵巢切除后,血清雌激素水平与对照组[（88±22vs403±59）pmol/L,P〈0.05]相比明显下降,左心室肌中两种雌激素受体表达均下降（P〈0.05）。给予雌激素后,血清雌激素水平上升为（3864±105）pmol/L,雌激素β受体表达增多（P〈0.05）,α受体表达下降（P〈0.05）。结论：雌激素改变卵巢切除后雌性大鼠心肌组织中雌激素受体的表达。     

中药复方联合三苯氧胺对实验大鼠乳腺增生组织及ER、PR表达的影响

目的探讨中药联合三苯氧胺(TAM)周期治疗大鼠乳腺增生的疗效及作用机理。方法将大鼠随机分为空白对照组、疾病模型组、三苯氧胺组、中药复方组、联合用药组5组,经雌二醇及孕酮刺激建立乳腺增生模型。光镜下观察乳腺组织病理学变化,免疫组化法测定雌激素受体(ER)、孕激素受体(PR)阳性表达强度。结果中药联合TAM治疗大鼠乳腺增生,可使乳腺小叶腺泡数明显减少、腺泡腔缩小及腺导管扩张程度减轻,ER、PR的阳性表达强度明显抑制,其疗效优于单纯中药和TAM组(P<0.05或0.01)。结论中药联合TAM治疗可降低实验大鼠乳腺组织中ER、PR含量,使其对雌激素的敏感性下降,对大鼠乳腺增生有明显的抑制作用。     

雌激素对去势雌鼠血清及膀胱组织e-NOS、AQPl含量的影晌

目的通过观察雌激素对去势雌鼠血清及膀胱e-NOS、AQPl含量的影响,进一步探讨雌激素替代治疗对绝经后尿道综合征的机制。方法成年健康SD雌性大鼠24只,随机分为空白组、对照组、尼尔雌醇组。摘除卵巢建立大鼠雌激素缺乏模型,灌胃给药4周后,用ELISA法测定血清及膀胱组织中e-NOS、AQP1的含量。结果尼尔雌醇组与对照组比较,血清e-NOS、AQPl含量显著增高（P〈0．05）．与空白组比较,差异无统计学意义（P〉0．05）。去势后膀胱e-NOS明显下降,补充雌激素后恢复正常水平,而AQPl去势前、后无显著改变,补充雌激素后亦无明显改变趋势。结论卵巢切除导致大鼠膀胱e-NOS发生下降,补充雌激素后恢复,而aQPZ无差异。e-NOS在绛．经后尿道综合征中起到直接作用．     

Molecular identification of potential selective estrogen receptor modulator (SERM) like properties of phytoestrogens in the human breast cancer cell line MCF-7

Numerous epidemiologic studies revealed that ethnic populations with higher dietary intake of phytoestrogens have the lowest incidence for breast cancer. The molecular mechanisms which may be responsible for this cancer protective action of phytoestrogens are so far only barely characterised. There are some hints that phytoestrogens may act like selective estrogen receptor modulators (SERMs) on the breast. For this reason we have investigated potential SERM-like properties of the phytoestrogens daidzein (Dai), coumestrol (Cou), and genistein (Gen) in the human breast cancer cell line MCF-7. Effects of these substances on progesterone (PR) and estrogen receptor alpha (ER) mRNA expression and estrogen receptor alpha protein levels were studied in comparison to estradiol (E2) and the synthetic SERMs raloxifene (Ral) and faslodex (ICI 182 780). PR mRNA expression was up-regulated after administration of Cou, whereas treatment with Dai and Gen induced only a faint increase. ER mRNA expression was down-regulated by Cou but not affected by Dai and Gen. The content of ER protein in the breast cancer cells was strongly decreased by Gen, only a faint reduction could be observed following administration of Cou, whereas administration of Dai slightly increases ER protein levels. In summary and in comparison to the effects observed after administration of E2, Ral, and ICI it turned out that Cou shows molecular properties which are very similar to an estrogen receptor agonist like E2, whereas the molecular properties of Gen are comparable to the SERMs ICI and Ral. These results clearly indicate that phytoestrogens differ significantly in regard to their molecular action on breast cancer cells and can be subdivided into distinct functional categories.     

Effects of genistein on the mammary gland proliferation of adult ovariectomised Wistar rats

The effects of phytoestrogens on the female breast are discussed controversially. On the one hand, epidemiological and experimental data provide evidence that dietary phytoestrogens may prevent the development of breast cancer. On the other hand, in breast cancer cell lines and tumour models isoflavone phytoestrogens have been demonstrated to stimulate the growth of estrogen-dependent breast cancer cells. To further investigate the molecular effects of genistein (Gen) on the mammary gland, we treated non-tumour bearing, ovariectomised female Wistar rats with this phytoestrogen either subcutaneously (10 mg/kg body weight) or orally (100 and 200 mg/kg body weight) for 3 days. Estradiol (E(2), 0.004 mg/kg s. c.) and ethynylestradiol (EE, 0.1 mg/kg per os) served as reference compounds. In the breast tissue, mRNA and protein expression of the progesterone receptor (marker for estrogenicity) and PCNA (marker gene for proliferation) were examined by quantitative real-time PCR, Western blotting and immunohistochemistry; the uterotrophic response was assessed also. Treatment with Gen per os or s. c. results in a small but significant stimulation of the uterine wet weight. In the mammary gland, Gen stimulates the expression of progesterone receptor (PR) but, in contrast to E(2), the isoflavone does not stimulate the expression of PCNA. These findings resemble recent data demonstrating a differential ability of Gen to induce uterine gene expression and uterine proliferation. Our data indicate that in non-malignant breast tissue short-term administration of Gen, in contrast to more potent estrogens like E(2), does not induce proliferation. Chronic stimulation of proliferation is believed to be a key mechanism during the development of breast cancer. The limited ability of Gen to stimulate proliferation in this tissue could be an indication for a limited carcinogenic potency of Gen in the breast. In further investigations it is important to identify molecular differences between healthy and malignant breast tissue which may explain the different sensitivity towards Gen treatment.     

凋膜止崩液对无排卵性功血大鼠子宫内膜组织ER、ERα及ERβ表达的影响

目的:探讨凋膜止崩液官腔靶向给药治疗无排卵性功能失调性子宫出血(功血)子宫内膜增生症的可能机制.方法:选取健康成年SD雌性大鼠48只,分为空白组、模型对照组、实验小剂量组和实验大剂量组,建立无排卵性功血子宫内膜增生症模型.凋膜止崩液官腔内给药后3d处死,留取子宫标本,采用免疫组化技术检测各组大鼠子宫内膜组织中雌激素受体(ER)、ERα及ERβ的表达水平.结果:模型对照组ER、ERα、ERβ的表达高于空白组,实验大、小剂量组ER、ERα、ERβ的表达低于模型对照组,差异均有统计学意义(均P＜0.001),实验大剂量组ER、ERα和ERβ水平低于实验小剂量组,差异有统计学意义(P＜0.01).结论:凋膜止崩液通过下调ER、ERα及ERβ阻断了雌激素发挥效应的通路,抑制子宫内膜的持续性增生从而治疗子宫内膜增生症.     

Tamoxifen and estrogen attenuate enhanced vascular reactivity induced by estrogen deficiency in rat carotid arteries

Recent clinical trials showed that estrogen usage in postmenopausal women did not affect coronary heart disease incidence, in contrast to several laboratory studies showing that estrogen decreased vascular reactivity. We speculated that, in some arteries, estrogen deficiency enhances endothelial function to compensate for the increased vascular smooth muscle reactivity. In this study, we examined the role of endothelium-derived vasoactive factors and the influence of in vivo estrogen and/or tamoxifen treatment on vascular reactivity of estrogen-deficient rats. Common carotid arteries were isolated from sham-operated (control), ovariectomized (Ovx), estrogen- or tamoxifen-treated Ovx rats, and Ovx rats co-treated with estrogen and tamoxifen. U46619 or phenylephrine induced similar contractions in endothelium-intact rings from all groups. Interestingly, removal of endothelium unmasked enhanced contractions in Ovx rats, which was prevented by estrogen, tamoxifen, or estrogen+tamoxifen treatment. Contractions to high K(+) were higher in both endothelium-intact and endothelium-denuded arteries from Ovx rats. Estrogen or tamoxifen treatment normalized high K(+)-induced contraction. A gap junction blocker, 18alpha-glycyrrhetinic acid, revealed enhanced contractions to U46619 in the absence or presence of l-NNA. Western blotting showed enhanced expressions of gap junctional connexin 43 in Ovx group. This study suggests that ovariectomy increases functional expression of gap junction-mediated endothelium-derived hyperpolarizing factor. Also, vascular effects of ovariectomy can be reversed by estrogen, tamoxifen or estrogen+tamoxifen treatment, suggesting that tamoxifen confers estrogenic effects in the vascular system.     

Estrogenic Endocrine-Disrupting Chemicals: Molecular Mechanisms of Actions on Putative Human Diseases

Endocrine-disrupting chemicals (EDC), including phthalates, bisphenol A (BPA), phytoestrogens such as genistein and daidzein, dichlorodiphenyltrichloroethane (DDT), and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), are associated with a variety of adverse health effects in organisms or progeny by altering the endocrine system. Environmental estrogens, including BPA, phthalates, and phytoestrogens, are the most extensively studied and are considered to mimic the actions of endogenous estrogen, 17β-estradiol (E2). Diverse modes of action of estrogen and estrogen receptors (ERα and ERβ) have been described, but the mode of action of estrogenic EDC is postulated to be more complex and needs to be more clearly elucidated. This review examines the adverse effects of estrogenic EDC on male or female reproductive systems and molecular mechanisms underlying EDC effects that modulate ER-mediated signaling. Mechanisms of action for estrogenic EDC may involve both ER-dependent and ER-independent pathways. Recent findings from systems toxicology of examining estrogenic EDC are also discussed.