大蒜素对小鼠脾细胞雌激素和雄激素受体影响的实验研究

本研究旨在探讨大蒜素对小鼠脾细胞雌激素和雄激素受体的影响。将健康ICR清洁小鼠,按性别随机分成三组实验组和三组对照组。实验组分别连续灌服大蒜素14、21和28 d,对照组同期分别灌服等量生理盐水。采用RT-PCR法检测雌性各组小鼠脾细胞雌激素受体(ERα和ERβ)的mRNA表达水平和雄性各组小鼠脾细胞雄激素受体(AR)的mRNA水平。实验结果显示,雌性小鼠连续灌服大蒜素第14和21 d时实验组ERαmRNA表达水平明显高于对照组(P0.05),第14 d时ERβmRNA表达水平明显高于对照组(P0.05)。雄性小鼠连续灌服大蒜素第14、21和28 d时实验组AR mRNA表达水平均明显高于对照组(P0.05)。综上提示大蒜素可增强小鼠脾细胞雌激素受体(ERα和ERβ)和雄激素受体(AR)mRNA的表达水平。     

大豆异黄酮对老龄大鼠雌激素水平和子宫雌激素受体表达的影响

目的探讨大豆异黄酮（soy isoflavones,SIF）对老龄大鼠雌激素水平和雌激素受体表达的影响。方法将40只16月龄雌性SD大鼠随机分为高、中和低剂量大豆异黄酮3个实验组（H-SIF,M-SIF和L-SIF）和1个对照组,每天分别按1005、0和25 mg/（kg.bw）剂量的大豆异黄酮进行灌胃,对照组灌服生理盐水。42 d后称重,股动脉放血处死动物收集血清、摘取子宫称重并冷冻保存。采用放射免疫法测定血清中雌二醇（E2）、睾酮（T）、泌乳素（PRL）、促黄体生成素（LH）和促卵泡激素（FSH）的含量;RT-PCR检测子宫雌激素受体α（ERα）基因的表达。结果与对照组相比,3个实验组血清中E2和PRL都升高,LH和FSH都降低,其中高剂量组差异有显著性（P〈0.05）;子宫重量无明显差异;子宫ERα表达呈下降趋势,但无统计学意义。结论大豆异黄酮能调节老龄大鼠血清中的雌激素水平,且有剂量依赖性;大豆异黄酮对老年大鼠子宫重量和子宫ERα的表达无影响。     

益坤宁对围绝经期大鼠下丘脑雌激素受体表达的影响

目的检测益坤宁对围绝经期大鼠下丘脑雌激素受体（estrogen receptor,ER）表达的影响,探讨益坤宁治疗围绝经期综合征的作用机制。方法采用自然衰老的围绝经期大鼠,随机分为围绝经期对照组、西药对照组和中药实验组;另取5月龄青年雌性大鼠做青年对照组。各组大鼠分别连续灌胃给予生理盐水、利维爱、中药益坤宁和生理盐水4周。通过免疫组化观察大鼠下丘脑ER蛋白的表达部位;Western blot定量检测大鼠下丘脑ERα和ERβ蛋白表达。结果ERα、ERβ蛋白主要表达于细胞胞质中。围绝经期对照组与青年组相比,ERa、E邓蛋白表达量明显下降（P〈0．05,P〈0．01）;中药实验组与围绝经期对照组比较,ERα、ERB蛋白表达量均明显增多（P〈0．05,P〈0．01）;西药对照组与围绝经期对照组相比,ERα、ERβ蛋白表达量增多（P〈0．05）。结论益坤宁通过提高围绝经期大鼠下丘脑组织中ERα、ERβ蛋白表达水平,使ER的合成增加,从而提高雌激素靶组织中ER的含量以增强雌激素的生物活性而发挥治疗作用。     

大豆异黄酮对雌鼠初情期等生殖性状的影响

目的通过研究大豆异黄酮对雌鼠初情期等生殖性状的影响,为母鼠的安全饲喂提供实验依据。方法本实验选用210只21日龄ICR雌鼠随机分成3组,饲喂含不同浓度大豆异黄酮（0 mg/kg、50 mg/kg、400 mg/kg）的饲料。每日检查母鼠阴道开张和阴道栓情况。采集母鼠45和65日龄时血清,利用ELISA方法检测血清中雌激素含量,并统计45、65日龄体重和子宫重。利用免疫组织化学方法检测子宫上皮雌激素受体的表达分布情况。结果雌鼠阴道开张的平均日龄分别为27.2 d、26.1 d和25.8 d,400 mg/kg组显著早于剂量为0mg/kg的对照组（P〈0.05）;雌鼠平均初次配种时间各组间差异不显著（P〉0.05）;饲养到45日龄和65日龄时400 mg/kg组雌鼠体重显著高于对照组（P〈0.05）。喂养8周后两个试验组在子宫间质细胞上雌激素受体的阳性细胞表达率显著高于对照组;在腺上皮中400 mg/kg组显著高于50 mg/kg组和对照组（0 mg/kg）;但是在腔上皮中,各组间受体的阳性细胞表达率差异不显著（P〉0.05）。结论饲喂每公斤含400 mg大豆异黄酮的饲料可以刺激初情期前母鼠阴道的开张,提高雌鼠的体重,降低65日龄时血清雌激素含量,并在饲喂8周后影响子宫间质和腺上皮上雌激素受体的表达分布;而饲喂每公斤含50 mg大豆异黄酮的饲料仅对小鼠45日龄体重及饲喂8周后子宫间质上雌激素受体的表达分布有所影响。     

低浓度臭氧对小鼠生长发育的影响

目的研究低浓度臭氧对小鼠生长发育的影响。方法实验组小鼠和对照组小鼠分别在15w紫外灯照射产生的低浓度臭氧环境下和正常无臭氧环境下连续饲养7周，观察其体重变化；阴道涂片镜检观察雌性小鼠的动情周期。7周后，取脑、心、肝、脾、肺、肾等组织标本，做组织病理学分析；利用全自动生化分析仪对部分血清生化指标进行测定，利用放免试剂盒对血清性激素水平进行测定。结果实验组雌性小鼠体重与对照组雌性小鼠差异无统计学意义，实验组雄性小鼠体重比对照组雄性小鼠轻；实验组雌性小鼠的动情周期与对照组雌性小鼠差异无统计学意义；实验组与对照组小鼠的组织切片均未发现异常；实验组小鼠的血清生化指标以及血清性激素水平与对照组小鼠差异无统计学意义。结论本实验中紫外灯照射产生的低浓度臭氧对昆明小鼠的生长发育基本无影响。     

雌激素对成年和老年雌性大鼠血管平滑肌细胞雌激素受体α表达的影响及其机制

目的探讨雌激素对成年和老年雌性大鼠血管平滑肌细胞(VSMC)雌激素受体α(ERα)表达的影响及其机制。方法采用RT-qPCR、Western blot及重硫酸盐修饰后测序(BSP)的方法,检测体外培养的3-4代成年(2-3月龄)和老年(≥20月龄)雌性SD大鼠VSMC在无或有生理剂量(10-10 mol/L、10-8 mol/L)17β-雌二醇(E2)存在下,ERα的表达及其启动子区CpG岛甲基化水平的变化。结果成年雌性大鼠VSMC无论在有或无雌激素存在时均表达ERα,且表达水平随雌激素浓度增加而上升。而老年雌性大鼠VSMC无论是在mRNA水平还是蛋白质水平几乎检测不到ERα表达,即使补充雌激素达最高生理剂量也无法诱导ERα的重新表达,其ERα基因启动区CpG岛呈现高水平甲基化。结论成年大鼠VSMC表达ERα,且生理剂量雌激素对其表达具有诱导作用。而老年大鼠VSMC ERα基因由于CpG岛已发生高度甲基化而抑制,生理剂量雌激素对ERα表达的诱导作用丧失。     

雌激素和孕激素对雌性小鼠甲状腺C细胞的影响

探讨降钙素在老年女性骨质疏松症发病中的作用 ,观察了雌、孕激素对雌性小鼠甲状腺 C细胞的影响。去卵巢小鼠 (OVX)分别肌肉注射苯甲酸雌二醇 (FB)、己烯孕酮 (HPC)和苯甲酸雌二醇加乙烯孕酮 (EB+HPC)两个月。用药剂量根据小鼠口龄和体重的不同而不同。采用免疫组织化学方法显示降钙素 (CT)阳性细胞并对其进行细胞形态计量学分析。结果显示 :(1)去卵巢小鼠与正常对照组相比 ,甲状腺 C细胞数目剧增 ,给予 EB、HPC、EB+HPC治疗的各组小鼠甲状腺C细胞数目与正常对照组水平相近。 (2 )各组小鼠甲状腺 C细胞的大小无明显改变 ,平均直径间无显著性差异 (P>0 .0 5 )。雌、孕激素缺乏可引起雌性小鼠甲状腺 C细胞增生。 C细胞增生也可能是雌激素缺乏引起的骨质疏松的后果     

人参茎叶皂甙对不同年龄大白鼠心肌细胞的影响

实验用3—5月龄青年鼠11只,18月龄老年鼠13只,各分对照与实验两组,实验组在基础饲料中加入参皂甙。三个月后取心肌制超薄切片,电镜观察。青年对照组与实验组心肌结构未见明显差别。老年对照组心肌细胞间胶元纤维增多。细胞内脂褐素数量增多,电子密度致密。核内异染色质在核边缘堆积且核边缘不规则,多凹陷。线粒体在肌原纤维间排列不整齐,线粒体嵴破裂以致消失。实验组细胞结构有所改善,表现在核边缘比较完整,异染色质较少,而且大部分线粒体嵴结构较清晰,脂褐素电子密度较低,文中井讨论了脂褐素的来源。     

UU1、UU3和UU4致昆明小鼠下生殖道致病性的研究

目的:研究解脲脲原体(Ureaplasma Urealyticum,UU)的第1血清型(UU1)、第3血清型(UU3)和第4血清型(UU4)对昆明小鼠下生殖道的致病性。方法:将100只昆明小鼠随机分为5组:空白组、雌二醇组、UU1组、UU3组和UU4组,每组20只;空白组和雌二醇组为对照组,UU1组、UU3组和UU4组为实验组;雌二醇组和实验组雌二醇化后1周,将UU1、UU3和UU4分别接种至UU1、UU3和UU4组小鼠泌尿生殖道,将液体培养基接种至空白组和雌激素组。2周后检测各组小鼠泌尿生殖道的IL-8、SIgA、TNF-α并取小鼠子宫组织行病理检查。结果:实验组小鼠UU接种都获得成功。IL-8、SIgA、TNF-α检测结果显示:实验组和对照组比较有显著性差异;UU4组和UU1组及UU3组比较有显著性差异;空白组和雌二醇组比较无显著性差异;UU1组和UU3组比较无显著性差异。病理显示:UU1、UU3和UU4组昆明小鼠子宫组织有炎细胞浸润,UU4组小鼠宫颈病变更明显。对照组小鼠子宫未见明显改变。结论:UU1、UU3和UU4可能导致昆明小鼠下生殖道产生病理反应;UU4可能短期致病力更强。     

UU1、UU3和UU4致昆明小鼠下生殖道致病性的研究

目的：研究解脲脲原体（Ureaplasma Urealyticum,UU）的第1血清型（UU1）、第3血清型（UU3）和第4血清型（UU4）对昆明小鼠下生殖道的致病性。方法：将100只昆明小鼠随机分为5组：空白组、雌二醇组、UU1组、UU3组和UU4组,每组20只;空白组和雌二醇组为对照组,UU1组、UU3组和UU4组为实验组;雌二醇组和实验组雌二醇化后1周,将UU1、UU3和UU4分别接种至UU1、UU3和UU4组小鼠泌尿生殖道,将液体培养基接种至空白组和雌激素组。2周后检测各组小鼠泌尿生殖道的IL-8、SIgA、TNF-α并取小鼠子宫组织行病理检查。结果：实验组小鼠UU接种都获得成功。IL-8、SIgA、TNF-α检测结果显示：实验组和对照组比较有显著性差异;UU4组和UU1组及UU3组比较有显著性差异;空白组和雌二醇组比较无显著性差异;UU1组和UU3组比较无显著性差异。病理显示：UU1、UU3和UU4组昆明小鼠子宫组织有炎细胞浸润,UU4组小鼠宫颈病变更明显。对照组小鼠子宫未见明显改变。结论：UU1、UU3和UU4可能导致昆明小鼠下生殖道产生病理反应;UU4可能短期致病力更强。     

苦瓜皂甙降血糖作用及其机制初探

将雄性Wistar鼠 (体重 1 80± 1 0g)随机分为正常对照组 (NC)、实验对照组 (EC)和两个实验组 (E1、E2 ) ,其中NC、EC组饮用普通水 ,E1、E2组饮用水中分别含有苦瓜皂甙 1 0 0mg·L 1和 2 0 0mg·L 1。饲养五周后 ,后三组动物禁食 2 4h后取血待测 ;取血完毕即开始进食 ,2 4h后再次取血待测。一周后杀死动物取肌肉、肝脏标本检测糖原水平。结果显示苦瓜皂甙虽不影响禁食大鼠血浆胰岛素含量 ,却可明显升高血糖和皮质醇水平 ;显著降低进食后大鼠血糖和胆固醇水平 ,并维持适度的皮质醇水平。肌糖原和肝糖原含量明显增加     

Senescence and steroid hormone receptor reactivities in accessory sex glands of elderly rats (Sprague-Dawley) following exogenous hormonal therapy

The aim of this study was to characterize the stromal and epithelial distribution of AR, ERα and ERβ reactivities in the different accessory sex glands of elderly rats and during strong hormonal changes. Ten month old male rats were divided into six senile groups and submitted to treatment: Senile/Control group (SC); Senile/Testosterone group (ST): Senile/Estrogen group (SE); Castrated group (CA); Castrated/Testosterone group (CT); Castrated/Estrogen group (CE). After a 30-day treatment, the prostatic ventral lobe (VL), dorsal lobe (DL) and coagulating gland (CG) samples were processed for immunohistochemistry and Western Blotting. The results showed that AR immunoreactivity was characterized in the epithelium of VL and DL in senile/control rats and senile rats submitted to exogenous hormonal therapy. AR reactivity in the coagulating gland was verified predominantly in the stromal cells in the different experimental groups. ERα reactivity occurred predominantly in the stromal compartment in all accessory sex glands. In the DL and CG, ERα immunoreactivities were intense in the groups which received testosterone (ST) and estrogen (SE). ERβ immunoreactivity in the CG was verified in the stromal compartment in the different experimental groups, showing a positive response to both increased testosterone and estrogen levels. ERβ reactivity, in the DL, was intensified in the stroma of senile rats with higher serum testosterone levels, and in senile rats with increased serum estrogen levels, especially in the glandular epithelium. Thus, the results revealed different distribution pattern of steroid hormone receptors in each one of the prostatic lobes in senescence, especially in the prostate dorsal lobe and coagulating gland, which is a fundamental factor due to the fact that major prostatic diseases occur in a later period of life.     

Influence of drinking water -- tap versus purified -- on embryo and fetal development in mice.

Pregnant CD-1 mice drank either chlorinated tap water obtained at NIEHS from the City of Durham, North Carolina municipal water supply or this same water passed in sequence through an organic removal cartridge and a demineralizer, followed by glass distillation. No significant overall differences in the reproductive status of pregnant mice were noted when the two groups were compared. Similarly, no significant overall influence on the incidence of malformed fetuses could be traced to the purity of drinking water. Month by month comparisons indicated that there were three statistically significant differences; all of which indicated improved performance for the mice that drank tap water. When the results for both groups were combined and a month by month comparison was made there was a significant difference in the pregnancy rate for February (68%) as compared with an overall pregnancy rate of 80% and a 79-93% spread for the other months, but the decrease was due to the low incidence of pregnancy in the group that drank the purified water.     

Differentiation between androgen and estrogen receptor mediated effects of testosterone on FSH using androgen receptor deficient (Tfm) and normal mice

The comparison of normal and androgen receptor (AR) deficient Tfm-mice allows distinction between AR mediated and estrogen receptor (ER) mediated effects of testosterone (T)--the latter after aromatization of T to estrogens--on serum and pituitary FSH. Normal male and female as well as Tfm mice were gonadectomized after 8 days and treated for 11 days with either T, estradiol (E2) or vehicle. Serum and pituitary FSH was determined by RIA for rat FSH. In Tfm mice T caused a suppression of serum FSH, indicating an ER mediated effect. Lower serum FSH levels after T in normal mice than Tfm mice indicate an additional AR mediated suppression. Lower serum FSH values in E2 treated Tfm than in T treated Tfm mice--where T acts only through ER--suggest two classes of estrophilic cells: one which aromatizes, thus being susceptible for both T and E2, and the other which does not aromatize. Only AR but not ER mediated T effects on pituitary FSH could be demonstrated.     

Repeat estradiol exposure differentially regulates protein expression patterns for estrogen receptor and E-cadherin in older mouse ovarian surface epithelium: Implications for replacement and adjuvant hormone therapies?

BackgroundEstrogen replacement therapy increases risk for ovarian epithelial cancer, a cancer of mainly older women, yet the response of older ovarian surface epithelium (OSE) to repeat estrogen exposure overtime has not been studied. We have previously reported significant reductions in estrogen receptor (ER) protein expression, particularly the ERβ1 isoform, in older mouse OSE following a single depot estradiol injection. The current study examined OSE from older mice following a single, and repeat estradiol injection, given 14 days apart over 28 days.MethodsCohorts of mice were sacrificed 48 hours following each estradiol injection, and at three other equidistant time points. Serum and ovarian tissue estradiol concentration was correlated to immunohistochemical and morphometric parameters used to identify evidence of OSE hyperplasia and hypertrophy. Using immunohistochemistry, E-cadherin expression was investigated in OSE 48 hours following both estradiol injections, while ERα and ERβ1 expression was examined in OSE following repeat estradiol exposure only.ResultsFirst exposure to exogenous estradiol resulted in OSE hypertrophy and hyperplasia, and high levels of E-cadherin expression. In contrast, repeat estradiol exposure resulted in no OSE hyperplasia or hypertrophy, low levels of E-cadherin expression, high ERα and reduced ERβ1 protein expression in OSE, and low stromal ERα expression. Blood and ovarian tissue estradiol levels following repeat estradiol injection were half those recorded after a first dose equivalent injection, but remained significantly elevated above controls.ConclusionRepeat estradiol exposure leads to accumulation of estradiol in ovarian tissue, differentially regulating protein expression patterns for E-cadherin in OSE and ER in OSE and stroma.     

ERα regulates lipid metabolism in bone through ATGL and perilipin

A decrease in bone mineral density during menopause is accompanied by an increase in adipocytes in the bone marrow space. Ovariectomy also leads to accumulation of fat in the bone marrow. Herein we show increased lipid accumulation in bone marrow from estrogen receptor alpha (ERα) knockout (ERαKO) mice compared to wild‐type (WT) mice or estrogen receptor beta (ERβ) knockout (ERβKO) mice. Similarly, bone marrow cells from ERαKO mice differentiated to adipocytes in culture also have increased lipid accumulation compared to cells from WT mice or ERβKO mice. Analysis of individual adipocytes shows that WT mice have fewer, but larger, lipid droplets per cell than adipocytes from ERαKO or ERβKO animals. Furthermore, higher levels of adipose triglyceride lipase (ATGL) protein in WT adipocytes correlate with increased lipolysis and fewer lipid droplets per cell and treatment with 17β‐estradiol (E2) potentiates this response. In contrast, cells from ERαKO mice display higher perilipin protein levels, promoting lipogenesis. Together these results demonstrate that E2 signals via ERα to regulate lipid droplet size and total lipid accumulation in the bone marrow space in vivo. J. Cell. Biochem. 114: 1306–1314, 2013. © 2012 Wiley Periodicals, Inc.     

17 beta-estradiol increases Ca(2+) influx and down regulates interleukin-2 receptor in mouse thymocytes

The influx of Ca(2+) across the T lymphocyte membrane is an essential triggering signal for activation and proliferation by an antigen. The aim of this study was to determine if Ca(2+) influx through estradiol receptor (ER) operated channels of Ca(2+) entry induced activation of lymphoid cells. Mouse thymocytes were incubated with 17 beta-estradiol (E) and in the presence or absence of the mitogen, phytohemagglutinin (PHA). Despite evidence of an enhanced binding of E to ER on thymocyte membranes, and an E dose-related influx of Ca(2+), there was a consistent down regulation of IL-2 receptor expression (P < 0.001). Incubation of thymocytes with PHA enhanced IL-2 receptor expression although the down regulatory effect of E was still evident. The results suggest that the Ca(2+) channel activated by E may have a down regulatory effect on the IL-2 receptor in thymus cells leading to the dampening of cell activation process.     

Sex hormone changes induced by the parasite lead to feminization of the male host in murine Taenia crassiceps cysticercosis

Female mice are more susceptible to Taenia crassiceps (TC) infection than males. However, after a month parasite load increases massively in both genders reaching thousands of parasites per host. The possibility of hormonal changes in the infected mice was envisaged. Sex hormones levels were assayed after different periods of infection, the parasites present in the peritoneal cavity were collected and gonads, uterus and seminal vesicles were weighed. In male mice, serum estradiol increased to levels 200 times their normal values whilst those of testosterone decreased 90% relative to controls. The weight of seminal vesicles was significantly diminished. Infected female mice also showed a slight increase in estrogen blood levels after 8 weeks of infection and the weight of the uterus was significantly increased relative to controls. Serum estradiol and testosterone were almost undetectable after gonadectomy. Cytokines such as IL-6 are capable of stimulating aromatase activity and we found that splenocytes from infected mice produced amounts of IL-6 higher than control as measured by ELISA. In conclusion T. crassiceps infection triggers a feminization process in the infected hosts. The gonads are required for the parasite to induce higher estrogen synthesis. IL-6 could be involved in the immunoendocrine mechanism used by the parasite to maintain a highly permissive environment for its rapid growth.     

Effect of estradiol and other endocrine factors on the level and dynamics of estrogen receptors in the liver cells of rats

The authors studied the effects of different doses of estradiol (E2) on the level of estrogen receptors (ER) in female rat hepatocytes and the dynamics of ER distribution between the cytosol and nuclear cell fractions and compared the changes in the ER level in the liver in different endocrine states of the body. It was shown that the ER level in hepatocytes was far lower than in uterine cells and drastically increased during puberation and after ovariectomy. It was also found that the ER level in hepatocytes was dependent on pituitary functions. After a single injection of E2, the ER level in cytosol descended and that in the nucleus rose. The degree of ER reduction in cytosol and increase in the nucleus correlated with the dose of E2. The dynamics of intercombined changes in the levels of cytosol and nuclear ER was studied at different time intervals following the injection of different doses of the hormone. Some essential organ-specific features of E2 reception in the liver were revealed. Different mechanisms of the control of ER content in the liver and uterus are postulated.     

May primary empty sella turcica be a cause of isolated ACTH deficiency? A case report and the review of related literature

Isolated ACTH deficiency is an uncommon cause of secondary adrenocortical insufficiency and accompaniment with primary empty sella has been reported in several cases. We present a case of isolated ACTH deficiency associated with empty sella. A sixty-two year old woman was admitted to our endocrine clinic with complaints of weakness, fatigue, weight loss, nausea, vomiting, and lack of appetite for about one month. Physical examination indicated orthostatic hypotension and epigastric tenderness. Laboratory investigations revealed hypoglycemia, hyponatremia and anemia, in addition low plasma cortisole and ACTH levels. Serum cortisole responses to short and prolonged ACTH stimulation were tested and partial and accurate responses were obtained, respectively. Plasma ACTH and serum cortisole levels failed to respond after intravenous injection of human corticotropin releasing hormone. Other hypophysial hormone levels were within the normal reference ranges. Although cranial and abdominal computerized tomography images were evaluated as normal, cranial magnetic resonance imaging of the pituitary gland revealed 'primary empty sella turcica'. Replacement therapy with methylprednisolon resulted in the improvement of hypoglycemia, hyponatremia and clinical symptoms. Based on these results, the patient was diagnosed as isolated ACTH deficiency and was scheduled for follow up by our outpatient clinic. Our report is consistent with other reports pointing out that primary empty sella may be responsible for pathogenesis of isolated ACTH deficiency.