自然发情与诱导发情小鼠子宫内膜中雌激素受体α表达的比较

目的从雌激素受体α(ERα)的角度探讨自然发情小鼠与诱导发情小鼠的子宫内膜上,雌激素受体α表达是否受内源雌激素的特异诱导而变化,两者之间是否存在差异。方法27只同日龄母鼠,根据处理方式的不同随机分为3个组:自然发情假孕组(对照组)、诱导发情处理假孕组和自然发情假孕第1天摘除卵巢组,3个组的小鼠在见栓后第4、6、8天分别取样后,采用免疫组织化学法观察小鼠子宫内膜中雌激素受体α的表达情况。结果免疫组化结果显示,3个处理组的小鼠子宫内膜上皮细胞核、胞质都有ERα存在,且主要表达在腺上皮;见栓第4、6、8天时,诱导发情处理组小鼠子宫内膜的ERα阳性率均显著高于自然发情组和自然发情第1天摘除卵巢组(P0.05);见栓第8天时,自然发情处理组小鼠子宫内膜中的ERα阳性率与自然发情第1天摘除卵巢组差异不显著(P0.05),但见栓第4、6天时两者阳性率差异显著,自然发情处理组小鼠子宫内膜中的ERα阳性率显著高于自然发情第1天摘除卵巢组(P0.05)。结论诱导发情处理的小鼠子宫内膜,其表面雌激素受体α表达显著高于自然发情小鼠,且两者都受其内源性雌激素的特异诱导而变化。     

雌激素和孕激素对不同发情方式小鼠子宫内膜中孕激素受体分布的影响

目的探讨雌（Estrogen,E2）、孕激素（Progesterone,P4）对同期发情与自然发情小鼠子宫内膜中孕激素受体（Progesterone receptor,PR）分布的影响。方法45只同日龄雌鼠,根据处理方式的不同随机分为5组：自然发情组（对照组）、同期发情组、卵巢摘除组、P4处理组和E2处理组,5组小鼠在见栓后第4、6、8天分别取样后,采用免疫组织化学法观察小鼠子宫内膜中PR的分布变化情况。结果免疫组织化学染色结果显示,5个处理组小鼠子宫内膜的三种细胞中都有PR存在;同期发情组小鼠子宫内膜中三种类型细胞PR的表达与自然发情组差异有显著性（P〈0.05）;P4处理组小鼠子宫内膜中三种类型细胞PR的表达在见栓第4、6天显著低于卵巢摘除组（P〈0.05）;E2处理组小鼠子宫内膜腺上皮和间质中PR在第4、6、8天时都显著高于卵巢摘除组（P〈0.05）,而在腔上皮中则显著低于卵巢摘除组（P〈0.05）。结论同期发情处理与自然发情小鼠的子宫内膜上PR的分布,都受E2和P4的特异诱导而变化。     

PMSG或FSH同期发情处理对小鼠卵巢、输卵管和子宫中孕激素受体分布的影响

目的探讨孕马血清促性腺激素（pregnant mare serum gonadotropin,PMSG）和促卵泡激素（folliclestimulating hormone,FSH）同期发情处理的小鼠卵巢、输卵管和子宫中孕激素受体（progesterone receptor,PR）分布的差异。方法 10只8周龄KM系雌鼠,随机分为PMSG和FSH两个组,第一次处理后48 h取其卵巢、输卵管、子宫进行固定,免疫组织化学法观察各组织中PR的分布。结果两组小鼠卵巢、输卵管和子宫内膜中均有PR表达;其中PMSG组初级卵泡、次级卵泡和输卵管的阳性率均显著高于FSH处理组（P〈0.05）;PMSG组各级卵泡的平均吸光度值均显著高于FSH处理组（P〈0.05）;PMSG处理组子宫内膜上皮与腺上皮中的阳性率显著高于FSH组（P〈0.05）,而基质和子宫内膜上皮中的平均吸光度值显著低于FSH组（P〈0.05）。结论 PMSG和FSH同期发情处理不同程度影响小鼠卵巢、输卵管和子宫中PR的表达分布;其中PMSG处理组小鼠卵巢、输卵管和子宫上皮中PR表达普遍显著高于FSH处理组。     

PMSG和FSH同期发情处理对小鼠子宫、卵巢和输卵管中雌激素α受体分布的影响

目的从雌激素α受体（estrogen receptorα,ERα）的角度探讨孕马血清促性腺激素（pregnant mareserum gonadotropin,PMSG）和促卵泡激素（follicle-stimulating hormone,FSH）处理小鼠的卵巢、输卵管和子宫中,ERα分布是否有显著性差异。方法 10只8周龄母鼠,随机分为处理方式不同的两个组：PMSG组和FSH组,两组均在处理第48小时取其卵巢、输卵管和子宫样固定,采用免疫组织化学法分别观察组织中ERα分布情况。结果免疫组化结果显示,两个处理组小鼠卵巢、输卵管和子宫内膜的细胞中都有ERα表达;PMSG处理组卵巢中的初级卵泡和成熟卵泡上ERα阳性率和平均吸光度均显著高于FSH处理组;FSH处理组的输卵管中ERα阳性率和平均吸光度均高于PMSG处理组;FSH处理组子宫基质和腺上皮细胞中ERα的阳性率显著高于PMSG组,其中PMSG组基质中的平均吸光度显著高于FSH组,而子宫内膜上皮细胞的阳性率和平均吸光度两处理组间差异无显著性。结论 PMSG和FSH同期发情诱导由于其特性可不同程度地影响小鼠卵巢、输卵管和子宫中ERα的分布,使之在不同组织中产生差异性变化。     

槐角苷对雌性小鼠的抗生育作用研究

以ICR小鼠为研究对象,研究了槐角苷对雌性小鼠的抗生育作用。受孕小鼠按照0 mg/kg·day、150 mg/kg·day、300 mg/kg·day、600 mg/kg·day浓度的槐角苷进行灌胃处理,检测其胚胎着床数,SEM检视子宫内膜结构以及免疫组化研究子宫雌激素受体(ERα)和孕激素受体(PR)的表达水平。结果表明:经600 mg/kg·day槐角苷处理的小鼠胚胎平均着床数显著降低(P0.01);第5天SEM结果显示600 mg/kg·day的处理组小鼠子宫内胞饮突的形成受阻,子宫内膜容受性发生显著改变;第4天、第5天及第6天的免疫组化结果显示600 mg/kg·day的处理组小鼠子宫ERα表达水平显著降低(P0.05),PR表达水平显著升高(P0.01)。研究表明,槐角苷能够通过调节子宫内ERα与PR的表达、影响胞饮突的形成、降低子宫内膜容受性等多种途径的相互作用从而导致小鼠胚胎着床的失败,显示出了显著的抗生育活性。     

槟榔碱对怀孕小鼠子宫ERα和PR的影响

以ICR怀孕小鼠为研究对象,探索了槟榔碱对怀孕小鼠的抗生育作用,重点研究了槟榔碱对小鼠子宫雌激素受体(ERα)和孕激素受体(PR)的影响。用0 mg/kg·day、5 mg/kg·day、10 mg/kg·day和20 mg/kg·day的槟榔碱分别处理怀孕小鼠后发现,槟榔碱处理能导致怀孕小鼠胚胎着床数量和子宫重量减少,并表现出剂量依赖性。当槟榔碱剂量为20 mg/kg·day时,槟榔碱处理组小鼠的胚胎数量减少为对照组的59%,子宫重量减少为对照组的77%。HE染色发现20 mg/kg·day槟榔碱处理能导致怀孕小鼠第5天子宫内腔张开和第6天脱膜失败。荧光定量和免疫组化结果显示,20 mg/kg·day的槟榔碱处理能显著上调ERα和PR在子宫组织中mRNA和蛋白的表达量(P0.05)。     

围植入期小鼠子宫内膜中整合素β3亚基的表达

目的 研究整合素β3亚基与胚泡植人的相关关系。方法 取未孕动情期、真孕及假孕3-6天小鼠子宫作切片,用免疫组织化学和图像分析法检测整合素β3亚基围植人期子宫内膜的表达状况和变化规律。结果 整合素β3亚基仅分布于子宫内膜腺上皮的细胞膜。在真孕组小鼠,受精后第4天,整合素β3亚基含量明显上升,第5天达到高峰,第6天又突然回落;假孕组小鼠,只有第5天时呈弱阳性表达,其余各天的表达均降至检测水平之下。结论 ①整合素β3亚基在真孕小鼠子宫内膜的表达与植人窗的开放相吻合,是子宫内膜接受性的客观标志。②启动整合素β3亚基表达变化的信号不仅受母体因素调控,还与胚泡刺激有关。     

乙烯利暴露对早孕小鼠子宫内膜蜕膜化的影响

该研究主要探讨乙烯利(ethephon,ETH)暴露对早孕小鼠子宫内膜蜕膜化的影响。从孕第一天开始每天经口灌胃给予CD1小鼠0、71.25、142.5和285 mg/kg ETH后,于孕第七天处死小鼠。观察子宫胚胎着床数量,记录子宫重量及体重,采用酶联免疫吸附实验(enzyme linked immunesorbent assay,ELISA)检测孕鼠血清雌孕激素水平,RT-PCR、Western blot和免疫组化法(immunohistochemistry,IHC)检测HOXA10、COX2和BMP2等蜕膜化标记分子的mRNA和蛋白表达水平。构建假孕小鼠体内人工诱导蜕膜化模型,RT-PCR检测蜕膜化标记分子HOXA10、COX2和BMP2的mRNA表达水平。结果表明,在285 mg/kg ETH暴露下,小鼠孕第七天子宫胚胎着床数量显著降低(P0.001),其绝对子宫重量和相对子宫重量均显著低于对照组(P0.001)。RT-PCR结果显示,285 mg/kg ETH暴露组BMP2和HOXA10 mRNA表达水平显著低于对照组(P0.001),COX2表达水平显著高于对照组(P0.001)。Western blot结果显示,与对照组相比,285 mg/kg ETH暴露组其子宫内膜蜕膜化标记分子HOXA10蛋白表达水平显著降低,COX2、MMP9、PR表达水平显著升高。IHC结果显示,与对照组相比,285 mg/kg ETH暴露组其子宫内膜蜕膜化标记分子HOXA10和BMP2蛋白表达水平显著降低。ELISA检测结果表明,285 mg/kg ETH暴露组其血清孕激素水平显著降低(P0.001)。体内人工诱导蜕膜化模型检测结果显示,ETH暴露组人工诱导蜕膜化反应程度降低,诱导侧子宫重量与非诱导侧子宫重量之比显著低于对照组,蜕膜化标记分子HOXA10和BMP2 mRNA表达水平显著降低。该研究结果表明,孕早期ETH暴露会损害小鼠子宫内膜蜕膜化。     

小鼠子宫内膜LIF基因表达与雌、孕激素的关系

白血病抑制因子（LIF）是一种多功能活性的糖蛋白,LIF基因在大多数妊娠第4天的小鼠子宫内膜进行着强烈的表达,然而LIF基因表达调控的机制目前尚不清楚。本实验对168只妊娠第4~5天的小鼠LIF基因表达和血清中雌、孕激素水平分别进行了检测,发现18只小鼠无LIF基因表达,其血清中雌、孕激素水平分别极显著（P＜0.01）和显著（0.01＜P＜0.05）低于其他表达的小鼠。提示：雌、孕激素对小鼠LIF基因表达过程中起着一定的作用,将为LIF基因表达调控机制的深入研究打下基础。     

原发不育症子宫内膜花生凝集素受体的研究

本文以PNA(花生凝集素)为探针,应用ABC亲和组化技术,对69例原发不育症患者的子宫内膜PNA受体进行了检测.结果表明,原发不育症子宫内膜的增生期、分泌期PNA受体的阳性率均显著低于年龄相似、有生育史的对照组子宫内膜的阳性率.不育症子宫内膜分泌欠佳的PNA受体阳性率显著低于不育症分泌期不同时期子宫内膜的阳性率.提示,这种差异可能与雌孕激素对两组子宫内膜作用水平的不同引起.     

Immunocytochemical analysis of estrogen and progestin receptors in uteri of steroid-treated and pregnant cats.

The purpose of this study was to determine the distribution of estrogen receptors (ER) and progestin receptors (PR) in specific uterine cell populations during various steroid hormone treatment regimens, and to determine if ER and PR distribution in the uterus is altered during implantation and the establishment of pregnancy in the cat. The tissues were processed for indirect immunocytochemical localization of receptors using specific monoclonal antibodies against ER and PR. ER were present in the nuclei of all epithelial cells and stromal fibroblasts in endometrium obtained from ovariectomized animals, whereas PR were only detectable in the nuclei of stromal fibroblasts. There was an apparent increase in the staining intensity and number of nuclei that stained positively for both ER and PR in all cell populations after 14 days of estradiol treatment. The administration of progesterone for 14 and 21 days, in the presence or absence of continuous estradiol, reduced the apparent intensity of staining and the number of nuclei staining positively for both ER and PR. ER were undetectable in the luminal epithelium, but remained in the glandular epithelial cells and stromal fibroblasts, whereas PR were only detectable in stromal fibroblasts. ER and PR localization in the endometrium obtained from estrus animals was similar to that observed in the estradiol-treated animals. A general decrease in intensity of staining for both ER and PR was evident by Day 5 postcoitus in pregnant animals. This decrease in intensity of staining continued until Day 12 postcoitus, when the distributions of ER and PR were similar to those observed in the ovariectomized estradiol-primed, progesterone-treated animals.(ABSTRACT TRUNCATED AT 250 WORDS)     

Cell-specific expression of estrogen-responsive genes in the uteri of cyclic, early pregnant and ovariectomized ewes

A single physiological dose of estradiol up-regulates estrogen receptor-alpha(ER), progesterone receptor (PR), glyceraldehyde 3-phosphate dehydrogenase (GAPDH), c-fos, cyclophilin, and actin mRNAs in the endometrium of ovariectomized ewes. Therefore, we hypothesized that these genes would be up-regulated by the preovulatory surge of estrogen which occurs on the evening of Day 15 in the estrous cycle of sheep. ER and PR mRNA concentrations increased between Day 15 and Day 1 in cyclic ewes in most endometrial epithelial cells, while GAPDH mRNA increased in epithelial and stromal cells in the deep endometrium. Day 15 pregnant ewes had lower expression of ER, PR, GAPDH, cyclophilin and actin genes. For ER and GAPDH mRNAs, the greatest reduction occurred in the superficial endometrium. Ovariectomized ewes demonstrated concentrations of ER, PR, and GAPDH mRNAs that were similar to those in the cyclic ewes. While concentrations of c-fos mRNA did not differ between groups, those of cyclophilin and actin mRNAs were lower in the pregnant and ovariectomized ewes. In conclusion, ER, PR and GAPDH gene expression rose during estrus in endometrial cells with the highest ER gene expression and were repressed in pregnant ewes in superficial endometrial cells with the greatest PR gene expression.     

Effect of plane of nutrition on endometrial sex steroid receptor expression in ewes

The effect of plane of nutrition on progesterone receptor (PR) and estrogen receptor alpha (ERalpha) expression in ovine endometrium was investigated. Rasa Aragonesa ewes (n=26) were fed diets to provide either 1.5 (Group C) or 0.5 (Group L) times the daily maintenance requirement and were slaughtered at Days 5 or 10 of the estrous cycle (Day 0=estrus). PR and ERalpha immunoreactivity were analyzed in eight endometrial cell compartments, defined by cell type and location. Group L had less PR immunostaining on Day 5 (P<0.05), which is consistent with lesser endometrial content of progesterone found in such animals. Most cell types of Group C had down regulation of PR at Day 10, but in Group L, this pattern was observed only in three cell compartments. The lesser PR contents found at Day 5 in Group L ewes may explain the lack of inhibition of PR. No effect of treatment or day of the estrous cycle was observed in ERalpha. Results indicate that endometrial PR is affected in a cell type, in specific manner, by plane of nutrition.     

孕酮对新生大鼠低氧缺血性脑损伤中MMP-3表达的影响

目的：研究孕酮（PROG）对新生大鼠低氧缺血后脑内基质金属蛋白酶3（MMP-3）表达的影响。方法：建立新生大鼠低氧缺血性脑损伤动物模型,伊文思兰（EB）染色和电镜观察新生鼠低氧缺血性脑损伤血一脑屏障的通透性改变;免疫印迹（Western blot）方法检测大脑皮层MMP-3表达。结果：电镜显示低氧缺血组血-脑屏障完整性明显破坏：EB染色结果表明低氧缺血组血-脑屏障通透性明显高于假手术组,差异极显著（P〈0．01）,孕酮组血-脑屏障通透性明显低于低氧缺血组,有显著性差异（P〈0．05）;Western blot结果显示低氧缺血组MMP-3蛋白表达显著高于假手术组（P〈0．01）;孕酮组MMP-3蛋白表达显著低于低氧缺血组（P〈0．05）。结论：孕酮通过减少MMP-3的表达,降低血一脑屏障的损伤,这可能是其发挥脑保护作用的机制之一。     

Estrus synchronization and artificial insemination of hair sheep ewes in the tropics

Hair sheep ewes (St. Croix White and Barbados Blackbelly) were used to evaluate 3 methods of estrus synchronization for use with transcervical artificial insemination (TAI). To synchronize estrus, ewes (n = 18) were treated with PGF2alpha (15 mg, im) 10 d apart, with controlled internal drug release (CIDR) devices containing 300 mg progesterone for 12 d (n = 18), or with intravaginal sponges containing 500 mg progesterone for 12 d (n = 18). On the day of the second PGF2alpha injection or at CIDR or sponge removal, sterile rams were placed with the ewes. Jugular blood samples were collected from the ewes at 6-h intervals until the time of ovulation, and daily for 16 d after estrus (Day 0). Plasma was harvested and stored at -20 degrees C until LH, and progesterone concentrations were determined by RIA. There was no difference (P>0.10) in time to estrus among the CIDR-, PGF2alpha- or sponge-treated ewes. All of the ewes in the CIDR group and 94.4% of the sponge treated ewes exhibited estrus by 36 h after ram introduction, while only 72.2% of PGF2alpha-treated ewes showed signs of estrus by this time (P<0.06). The time from ram introduction to ovulation was not different (P>0.10) among the CIDR-, PGF2alpha- or sponge-treated ewes. The time to the preovulatory LH surge was similar (P>0.10) among CIDR, PGF2alpha and sponge treated ewes. Progesterone levels through Day 16 after the synchronized estrus were not different (P>0.10) among treatment groups. Hair sheep ewes (n = 23) were synchronized using PGF2alpha and bred by TAI using frozen-thawed semen 48 h after the second injection. The conception rate to TAI was 2/23 (8.7%) and produced 3 ram lambs. In a subsequent trial, 17 ewes were synchronized with CIDR devices and bred by TAI using frozen-thawed semen 48 h after CIDR removal, resulting in a conception rate of 52.9% (9/17). It is possible to synchronize estrus in hair sheep using either CIDRs, sponges or PGF2alpha. Even though there were no significant differences in the timing of ovulation or the LH surge among the treatment groups, a higher conception rate was achieved in ewes synchronized with CIDR devices during the second trial. This may reflect an increase in the skill level of the TAI technician.     

Synchronization of estrus with PGF2 alpha administered 18 days after a progesterone treatment in lactating dairy cows

In a previous study we showed that estrus synchronization with 2 treatments of PGF2 alpha 13 d apart reduced conception rate at the synchronized estrus and that this reduction occurred mainly in cows in the early luteal phase at the second PGF2 alpha treatment. The objective of the present study was to determine the efficacy of a synchronization regimen in which PGF2 alpha was administered during the mid- to late-luteal phase to cows that had previously been synchronized with progesterone. Spring-calving cows from 6 dairy herds were used in this study. On Day -32 (Day 1 = the start of the breeding season), cows that had calved 2 or more weeks ago were randomly assigned to a synchronization (S, n = 732) or control (C, n = 731) group. Cows in Group S were treated with an intravaginal progesterone device (CIDR) for 12 d from Day -32 to Day -20, while those in Group C were left untreated. Similar percentages of cows in Group S (80.6%) and C (82.9%) had cycled by Day -7. The CIDR treatment synchronized the onset of estrus, resulting in 92.9% of cows in estrus being detected within 7 d after CIDR removal. Cows in Group S that had cycled by Day -7 were treated with PGF2 alpha (25 mg, i.m., Lutalyse) on Day -2. Cows in both groups that were anestrous on Day -7 were treated with a combination of progesterone and estradiol benzoate (EB) to induce estrus and ovulation (CIDR and a 10 mg EB capsule on Day -7, CIDR removal on Day -2, and injection of 1 mg EB 48 h after CIDR removal). The PGF2 alpha treatment synchronized the onset of estrus in 87.5% of the cows. Group S and C cows had similar conception rates to first (61.0 vs 58.3%) and second (58.4 vs 60.9%) AI; similar pregnancy rates over the AI period (82.8 vs 79.2%) and over the whole breeding season (91.9 vs 90.6%); and required a similar number of services per pregnancy to AI (1.7 vs 1.8). The interval from the start of the breeding season to conception for cows conceiving to AI or to combined AI and natural mating was shorter (P < 0.001) by 5.7 and 6.2 d, respectively, for the Group S cows. It is concluded that the treatment regimen tested in the present study achieved satisfactory estrus synchronization, had no detrimental effect on fertility at the synchronized estrus, and shortened the interval from start of the breeding season to conception.