雌激素α和β受体在大鼠结膜上皮和Henle腺上的分布

目的阐明雌激素α、β受体在大鼠结膜上皮和Henle腺的分布。方法选择22例青春期SD雌性大鼠上眼睑标本,采用免疫组织化学方法对结膜上皮和Henle腺雌激素α、β受体进行检测;同时采用放射免疫分析技术检测大鼠血清雌二醇的浓度。结果22例大鼠中,19例大鼠的结膜上皮雌激素α受体呈阳性表达,17例大鼠的结膜上皮雌激素β受体呈阳性表达。18例大鼠Henle腺深部的基底细胞雌激素α受体呈阳性表达,17例大鼠的Henle腺深部的基底细胞雌激素β受体呈阳性表达。在所有的大鼠中,Henle腺的杯状细胞雌激素α、β受体均呈阴性表达。雌激素α、β受体在结膜上皮阳性表达率间比较,差异无显著性(P>0.05);雌激素α、β受体在Henle腺深部的基底细胞阳性表达率间比较,差异亦无显著性(P>0.05)。正常大鼠血清雌二醇水平经检测含量为(22.13±3.54)ng/L。结论雌激素及其受体对大鼠结膜上皮、Henle腺深部的基底细胞具有重要的调控作用,但对Henle腺的杯状细胞分泌黏液功能无直接调节作用。     

持续光照对大鼠卵巢褪黑素受体表达的影响及与卵泡发育的相关性

目的:研究持续光照对青春期前雌性大鼠卵巢褪黑素受体的表达及与卵泡发育的相关性。方法:雌性大鼠随机分成对照组和24 h持续光照组,酶联免疫吸附法(ELISA)检测血清中褪黑素、雌二醇水平,免疫组织化学法观察卵巢组织褪黑素受体(MR)的表达,RT-PCR法检测卵巢组织中雌激素受体α、β(ERα、ERβ)的mRNA的表达量。结果:与对照组相比,24 h持续光照组阴门开启时间提前,动情期延长,血清褪黑素显著下降,雌二醇水平显著增高,卵巢组织中MR表达下降,ER表达增高。结论:持续光照可以抑制青春期前雌性大鼠褪黑素及其受体表达,并促进卵泡发育和卵巢雌激素受体的表达,这可能与女性性早熟的发病机制相关。     

禁止交配下调成年雌性大鼠乳腺组织雌二醇水平及雌激素受体α的表达

目的 研究禁止交配对成年雌性大鼠乳腺组织雌二醇(E2)水平、雌激素受体α(ERα)表达的影响.方法 12只成年雌性SD大鼠随机分为对照组(交配组)和禁止交配组,实验第61天终止实验后在间情期采集标本,光镜下观察各组大鼠乳腺组织结构,透射电镜下观察乳腺组织超微结构;采用放射免疫法检测外周血及乳腺组织E2浓度;采用免疫组织化学染色和Western blotting技术检测乳腺组织ERα的表达变化. 结果禁止交配60d后,成年雌性大鼠乳腺组织E2水平下降,与对照组比较P<0.05;同时乳腺组织上皮细胞、基质细胞细胞核ERα表达下调,与对照组比较P<0.05;光镜、电镜下各组大鼠乳腺组织未见明显异常. 结论禁止交配可下调雌性大鼠乳腺组织E2水平和ERα的表达.     

雌激素对大鼠心脏雌激素受体α和β表达的影响

目的研究雌激素受体（ER）α和β在新生、成年和卵巢切除大鼠心脏的表达。方法采用半定量逆转录聚合酶链反应（RT-PCR）和Western blot方法,分析新生和成年大鼠心脏各部的ERα和βmRNA及ER蛋白质的表达,探讨不同剂量的17β雌二醇对卵巢切除大鼠心脏ERα和βmRNA、ER蛋白质的表达调控及对心房、心室重量和它们与体重比变化的影响。结果ERα mRNA在新生大鼠心脏各部表达较低,而成年大鼠心脏各部ERα mRNA的转录水平提高3～20倍,这种差别体现在ERα蛋白质的表达水平不同。反之新生大鼠心脏ERβ mRNA表达水平较高,而成年大鼠心脏ERβ mRNA的表达降低2～7倍。卵巢切除减少成年大鼠心房ERαmRNA和蛋白质的表达,减轻心房重量及心房／体重比;17肛雌二醇替代却能反转这些改变,同时随着血浆17β-雌二醇水平的增高,这种影响在一定程度上呈剂量依赖关系。卵巢切除和178一雌二醇替代并不改变心脏和心室／体重比,也不影响ERβ mRNA的表达。结论雌激素受体α和β在新生和成年大鼠心脏的表达存在差别,ERβ可能在大鼠心脏的发育过程中起重要作用,ERα足雌激素对成年大鼠心脏调控的优势受体;雌激素的主要靶器官是心房,且通过其受体的介导促进心房肌细胞的增生。     

卵巢切除对大鼠肾上腺雌激素受体亚型表达的影响

目的通过对去卵巢大鼠雌激素受体(ER)亚型在肾上腺表达的观察,研究去卵巢对大鼠肾上腺功能影响的机制。方法选择成年Wistar大鼠20只,10只为假手术组,10只为模型组(去卵巢组),6周后处死大鼠,分离摘取肾上腺,称重后常规石蜡包埋切片,分别进行ERα、ERβ抗体的免疫组织化学染色,并取动物血清测量雌二醇(E2)和促卵泡生成素(FSH)的含量。结果1.ERα与ERβ在大鼠肾上腺均有表达,主要分布在肾上腺皮质部,以球状带和束状带为主,网状带有少量分布,ERβ的表达强于ERα。2.ERβ在大鼠模型组的表达强于假手术组,有统计学意义,ERα在模型组的表达略强于假手术组,但无统计学意义。3.卵巢切除后,大鼠血清FSH升高,E2下降,与假手术组相比,有显著差异。结论卵巢切除可影响雌激素受体在大鼠肾上腺的表达,尤其是对ERβ表达的影响较为显著。卵巢切除能够减少大鼠血清雌激素的分泌,使垂体反馈性的增加FSH的分泌。     

异补骨脂素对人晶状体上皮细胞雌激素受体表达的上调作用

目的: 研究人晶状体上皮细胞(HLECs)雌激素受体(ER)表达的情况及中药植物雌激素异补骨脂素(ISR)对ER表达的影响,为中药植物雌激素防治老年性白内障提供实验依据。方法: 本研究将ISR与HLEC共同孵育,以雌二醇(E₂)作为阳性对照,再用H₂O₂对HLECs造成氧化损伤后,采用流式细胞术检测不同浓度的ISR对ER蛋白表达的影响。结果: 正常HLECs存在ERα、β的表达;H₂O₂组HLECs ERα、β表达明显下降,与空白组比较差异显著(P<0.01);E₂和ISR高、中、低浓度组HLECs的ERα、β表达明显升高,与H₂O₂组比较差异显著(P<0.01);且随ISR浓度的增高ERα、β表达逐渐升高,呈明显的浓度依赖关系。 结论: E₂、ISR能上调经H₂O₂处理的HLECs的ERα、ERβ表达,并呈明显的浓度依赖关系,其抗氧化损伤作用,可能是通过上调ERα、ERβ表达实现的。     

APP17肽对卵巢去势大鼠海马神经元退行性变的作用

目的 通过观察APP17肽对卵巢去势大鼠海马神经元退行性变的作用。证实APP17肽对神经元退行性变的改善作用。方法 雌性大鼠，行双侧卵巢切除手术造模。并于6周后皮下注射APP17肽进行治疗。手术后12周行水迷宫实验，至13周取血并行灌注固定，取脑海马组织作超薄切片电镜分析，并作冰冻切片进行雌激素受体（ERα），神经生长因子（NGF）及脑源性神经生长因子（BDNF）免疫组织化学染色，并用全自动化学发光免疫分析仪测定血清雌二醇水平。结果 卵巢去势组大鼠血清雌二醇水平降低。水迷宫检测卵巢去势组大鼠较正常组游完全程所须时间明显延长，错误反应次数明显增多，海马神经元超微结构出现明显损害；海马光镜观察发现ERα表达增多，NGF及BDNF表达减少，使用APP17肽治疗后并不改变血清雌二醇水平，但能明显改善海马神经元超微结构的损害，使上述有关蛋白质的表达恢复到正常水平。结论 APP17肽不同通过改变雌激素水平来发挥改善神经元功能减退的作用。而可能是通过神经营养因子起作用。     

雌激素对去势大鼠髁突软骨雌激素受体基因表达的影响

目的探讨不同浓度的雌二醇(E2)对去势大鼠髁突软骨雌激素受体(ER)基因ERα和ERβ表达的影响。方法将SD大鼠分为对照组(control)、假手术组(sham)、去势组(OVX)、及时和延迟替代治疗组(OVX/17β-E2),每天静脉注射不同浓度雌二醇(5×10、5×10~2和5×10~3μg/kg),阴道上皮脱落细胞涂片监测雌激素水平,用Western blot和real-time PCR法检测大鼠颞下颌关节髁突软骨ERα和ERβ的蛋白及mRNA表达。结果去势大鼠阴道上皮细胞MI指数180/12/8,表明雌激素水平低下(P0.05)。及时补给生理浓度雌二醇(5×10~2μg/kg)能维持髁突软骨ER于正常范围,延迟补给生理浓度雌二醇不能恢复或改善ER表达;及时和延迟补给高浓度(5×10~3μg/kg)及低浓度(50μg/kg)E2均抑制ER蛋白和mRNA合成(P0.05),且高浓度效应显著。结论及时补充合理浓度的雌激素对去势大鼠颞下颌关节的正常生理功能具有重要意义。     

子宫肌瘤雌激素受体亚型mRNA的表达

目的　探讨雌激素受体亚型的表达与子宫平滑肌瘤发生、发展的关系。方法　将60例子宫肌瘤患者分为AB两组, A组32例<48岁,B组28例>48岁,应用原位杂交技术测定平滑肌瘤组织中雌激素两种受体亚型mRNA的表达,取肿瘤周 围正常的平滑肌作对照。结果　子宫肌瘤组织中ERαmRNA的表达明显高于正常的平滑肌组织,而ERβmRNA的表达降 低;B组ERαmRNA在子宫平滑肌瘤组织中的表达较A组呈下降趋势,而ERβmRNA的表达呈上升趋势。结论　子宫肌瘤的 发生发展与ERαmRNA的高表达有关,亦与ERβmRNA表达减少有关。肌瘤的萎缩可能与ERβmRNA表达的升高趋势及 ERαmRNA表达的下降趋势均相关。     

雌激素及其受体对低氧性肺血管重建的作用

 目的：探讨雌激素及其受体对低氧性肺血管重建的作用。 方法:采用间断常压低氧法建立大鼠低氧性肺动脉高压模型。将30只雄性SD大鼠随机分成：对照组、雌激素预防组、低氧组、雌激素治疗组、治疗对照组。以右心导管法测定平均肺动脉压(mPAP)；测定右心室肥厚指数［RV/(LV+S)］；图像分析技术测定肺小血管管壁厚度；用放射免疫法测定血清17-β雌二醇浓度。用RT-PCR方法检测肺组织中ERβ mRNA的表达，用免疫组化染色法检测肺小血管壁的ERβ蛋白表达。 结果:①低氧组、治疗对照组大鼠mPAP、RV/(LV+S)、WT%和WA%均高于对照组；雌激素预防组、雌激素治疗组大鼠mPAP、RV/(LV+S)、WT%和WA%均低于低氧组、治疗对照组。②低氧组、治疗对照组大鼠ERβ mRNA表达量和ERβ蛋白表达与正常对照组无差异。经雌激素干预后，雌激素预防组、雌激素治疗组的ERβ mRNA表达和ERβ蛋白表达显著高于低氧组、治疗对照组。 结论:17-β雌二醇能有效降低低氧性肺动脉高压大鼠的肺动脉压力、阻抑右心室肥厚,对低氧性肺动脉高压血管重建具有一定的预防和治疗作用，其作用可能是通过雌激素受体β实现的。     

Extremely low-frequency electromagnetic fields differentially regulate estrogen receptor-α and -β expression in the rat olfactory bulb

Recently, the effects of extremely low-frequency electromagnetic fields (ELF EMF) on biological systems have been extensively investigated. In this report, the influence of ELF EMF on olfactory bulb (OB) estrogen receptor-α (ERα) mRNA and -β (ERβ) mRNA expression was studied by RT-PCR in adult female and male rats. Results reveal for the first time that ELF EMF exerted a biphasic effect on female OB ERβ mRNA gene expression, which increased during diestrous and decreased during estrous. We did not observe any influence of ELF EMF on female OB ERα mRNA expression. Our data demonstrate a fluctuating pattern of ER-α and -β mRNA expression in the female OB throughout the phases of the estrous cycle in non-ELF EMF-exposed animals. Thus the highest ERα expression was observed in diestrous and the lowest in proestrous. The pattern of ERβ mRNA was less variable, the lowest expression was observed in diestrous. ER-α mRNA and -β mRNA expression level in the male OB did not exhibit any variation either in ELF EMF-exposed or non-ELF EMF-exposed animals. In summary, ELF EMF modulate ERβ gene expression in the OB of female adult rats but not in males.     

Immunohistochemical detection of estrogen receptor �� in alveolar bone cells of estradiol-treated female rats: possible direct action of estrogen on osteoclast life span

The role of estrogen in bone resorption has been specifically related to the effect of estrogen on the signalling pathway that inhibits the formation of osteoclasts. However, osteoclast apoptosis and a significant reduction in the number of these cells have been observed in the alveolar bone of female rats treated with estradiol. In the present study, the expression of estrogen receptor β (ERβ) in the cells of alveolar bone was evaluated in estradiol‐treated and ‐untreated female rats. In order to test the possible direct action of estrogen on osteoclasts, the relationship between apoptosis and ERβ expression in these cells was also analysed. The animals received estradiol for 14 days and the alveolar bone fragments were embedded in paraffin for the quantification of tartrate‐resistant acid phosphatase‐positive osteoclasts. The expression of ERβ and apoptosis in the osteoclasts were evaluated by ERβ immunohistochemistry and Terminal deoxynucleotidyl transferase‐mediated dUTP Nick‐End Labelling (TUNEL) methods, respectively. To confirm osteoclast death by apoptosis, these cells were analysed under transmission electron microscopy. Some osteoclasts from estradiol‐treated animals were found to be undergoing apoptosis and the number of tartrate‐resistant acid phosphatase‐positive osteoclasts was significantly reduced. ERβ immunolabelling was observed in the cytoplasm and nuclei of active osteoblasts, osteocytes and osteoclasts in both groups, suggesting a direct participation of estrogen on alveolar bone cells. However, following estradiol treatment, a strong ERβ immunolabelling was often observed in the TUNEL‐positive osteoclasts. Therefore, these results indicate that, in addition to the other signalling pathway, the reduction of alveolar bone resorption is also related to a direct action of estrogen on osteoclasts, promoting apoptosis in these cells, via ERβ.     

卵巢切除大鼠睑板腺雌激素α、β受体表达与组织病理学改变的相关性

目的 探讨卵巢切除后大鼠雌激素α、β受体(Erα、Erβ)表达与睑板腺组织病理学改变的相关性.方法 取48只成年SD雌性大鼠,随机分为正常对照组、假手术组和去势组(去卵巢组),每组16只,12周后处死大鼠,免疫组织化学检测睑板腺组织Erα和Erβ表达变化,光学显微镜和透射电子显微镜观察睑板腺组织学改变,应用放射免疫方法检测血清雌二醇水平.结果 去势组Erα表达与正常对照组间差异无统计学意义(P>0.05 ),但Erβ表达弱于正常对照组(P < 0.01).正常对照组Erα /Erβ表达比值为1.06,而去势组Erα/Erβ表达比值为1.83.光镜下观察去势组睑板腺腺泡组织结构排列紊乱,细胞体积变小,腺泡呈萎缩状态.电镜下观察去势组睑板腺腺泡上皮的基底细胞体积变小,细胞核形态变异多样,呈不规则形、肾形或长杆状; 过渡细胞的体积变小,脂滴减少,线粒体肿胀,线粒体嵴结构破坏,呈空泡化或云雾状改变,并可见大量嗜锇物质形成的圆形或不规则形电子致密的板层小体或髓鞘样结构.结论 卵巢切除后睑板腺组织病理学改变可能与Erβ表达减弱或Erα/Erβ比值失调有关.     

Roles of estrogen receptors alpha and beta in sexually dimorphic neuroprotection against glutamate toxicity

Although most agree that 17β-estradiol is neuroprotective via a variety of mechanisms, less is known about the role that biological sex plays in receptor-mediated estradiol neuroprotection. To address this issue we isolated primary cortical neurons from rat pups sorted by sex and assessed the ability of estradiol to protect the neurons from death induced by glutamate. Five-minute pretreatment with 10–50 nM 17β-estradiol protected female but not male neurons from glutamate toxicity 24 h later. Both estrogen receptor alpha (ERα) and estrogen receptor beta (ERβ) are expressed in these cultures. Experiments using an ERα selective agonist or antagonist indicate that this receptor is important for neuroprotection in female cortical neurons. The ERβ selective agonist conveys a small degree of neuroprotection to both male and female cortical neurons. Interestingly, we found that 17α estradiol and the novel membrane estrogen receptor (mER) agonist STX, but not bovine serum albumin conjugated estradiol or the GPR30 agonist G1 were neuroprotective in both male and female neurons. Taken together these data highlight a role for ERα in sexually dimorphic neuroprotection.     

Estrogen Alleviates Acetic Acid-Induced Gastric or Colonic Damage via Both ERα- and ERβ-Mediated and Direct Antioxidant Mechanisms in Rats

In order to demonstrate the possible protective effects of estrogen receptor (ER)-α and ERβ receptor subtypes in the pathogenesis of colonic and gastric oxidant damage, experimental ulcer and colitis were induced by acetic acid, and the animals were randomly divided as colitis, ulcer, and their corresponding non-ulcer and non-colitis control groups. Each group of rats was treated intramuscularly with the vehicle, selective ERα agonist propylpyrazole-triol (1 mg/kg), ERβ agonist diarylpropionitrile (1 mg/kg), non-selective ER agonist 17β estradiol (E2; 1 mg/kg), or E2 plus non-selective ER antagonist ICI-182780 (1 mg/kg). The results revealed that induction of ulcer or colitis resulted in systemic inflammation as assessed by increased levels of plasma TNF-α and IL-6 levels. In both tissues, the presence of oxidant damage was verified by histological analysis and elevated myleoperoxidase activity. In the colitis and ulcer groups, both ER agonists and the non-selective E2 reversed the oxidative damage in a similar manner. These findings indicate that estrogen acts via both ERα- and ERβ-mediated and direct antioxidant mechanisms, where both ER subtypes play equal and efficient roles in the anti-inflammatory action of estrogen, in limiting the migration of neutrophils to the inflamed tissue, reducing the release and activation of cytokines and thereby alleviating tissue damage.     

肺气肿模型大鼠肾组织水通道蛋白2的表达及机制研究

目的: 观察肺气肿模型大鼠肾组织AQP2表达以及血中ET、IL-1β、TNF-α水平的变化,探讨在体液代谢过程中肺与肾脏之间关系。方法: 用气管内注入脂多糖加熏香烟方法复制肺气肿大鼠模型,应用免疫组化、Western blotting、RT-PCR方法测定肾组织AQP2蛋白及mRNA表达的变化,应用放免法测定血中、肺组织匀浆中ET、IL-1β及TNF-α含量。结果: 与对照组比较,肾组织AQP2蛋白及mRNA表达增强(P<0.01);同时模型组血、肺组织ET、IL-1β及TNF-α含量明显升高(P<0.01)。结论: 肺气肿模型大鼠肾组织AQP2表达上调;其机制可能与血中ET、IL-1β、TNF-α含量升高有关。     

ERα和ERβ在正常子宫内膜、异常增生子宫内膜和子宫内膜癌组织中的表达及意义

目的探讨雌激素受体（ER）亚型α和β mRNA及蛋白的异常表达与子宫内膜癌的发生发展的关系。方法应用半定量逆转录聚合酶链反应（RT—PCR）和免疫组织化学链霉菌抗生物素蛋白-过氧化酶连接法（SP法）,检测18例正常子宫内膜组织、29例异常增生组织和50例子宫内膜癌组织中ERα、ERβ mRNA及蛋白的表达。结果①ERα mRNA在内膜癌组织中的阳性表达率和阳性表达水平明显低于异常增生内膜组织及正常内膜组织,三者比较有显著性差异（P〈0．01）,ERα蛋白在这三组间的表达率变化相同（P〈0．05）。②ERβ mRNA在内膜癌组织中的阳性表达率和阳性表达水平明显低于增生性内膜组织及正常内膜组织,三者比较有显著性差异（P〈0．01）,ERβ蛋白在这三组间的表达率变化相同（P〈0．01）。③ERα和ERβ mRNA平均表达水平比值（ERα／ERβ）在内膜癌组低于异常增生子宫内膜组织和正常子宫内膜组（P〈0．05）。结论ERα和ERβ表达的下调可能与子宫内膜癌的发生有关,在这个过程中ERα的下调起的作用大于ERβ。