白血病抑制因子在妊娠输卵管黏膜中的表达

目的探讨白血病抑制因子(LIF)与输卵管妊娠的关系。方法选择输卵管妊娠组25例、正常输卵管组28例及正常宫内早孕组30例,采用免疫组织化学染色技术及半定量病理图像分析系统检测输卵管妊娠黏膜组织、正常输卵管黏膜组织及正常宫内早孕组子宫蜕膜组织中LIF的表达。结果 LIF阳性表达在正常宫内早孕组最高,在输卵管妊娠组中较高,在正常输卵管组中最低,三组两两比较差异均有有统计学意义(P<0.05)。结论 LIF可能参与了输卵管妊娠发病过程,且与输卵管妊娠缺乏蜕膜化反应有关。     

妊娠早期小鼠卵巢,输卵管及子宫内诱导型一氧化氮合酶的分析

目的 了解胚泡着床前后妊娠昆明系小鼠卵巢、输卵管及子宫内诱导型一氧化氮合酶（ｉｎｄｕｃｉｂｌｅ ｎｉｔｒｉｃ ｏｘｉｄｅ ｓｙｎｔｈａｓｅ,ｉＮＯＳ）的分布。方法 免疫细胞化学ＬＳＡＢ法。结果 妊娠２～５ｄ小鼠的卵巢内,黄体细胞上有ｉＮＯＳ的阳性表达;输卵管粘膜上有ｉＮＯＳ的分布,肌层则为阴性;妊娠２、３ｄ的小鼠子宫内,阳性标记主要出现在子宫内膜上皮以及子宫内膜中的了宫腺上皮,内膜基质细胞为阴性;妊娠４ｄ的子宫内,子宫腺及蜕膜部分均有ｉＮＯＳ的分布,妊娠５ｄ时,在小鼠胚泡的表面也检测到了ｉＮＯＳ的存在。结论 小鼠胚泡着床前后,在其卵巢、输卵管及子宫内均有ｉＮＯＳ的存在,提示ｉＮＯＳ在小鼠胚胎早期发育及着床过程中起作用。     

ATF4基因在小鼠动情周期中子宫内膜的表达

目的探讨转录激活因子4(ATF4)在小鼠动情周期中子宫内膜的表达规律。方法采用RT-PCR、免疫组织化学和Western blot法分别检测小鼠子宫内膜中ATF4 mRNA和蛋白在动情前期、动情期、动情间期和动情后期的表达规律。结果 ATF4 mRNA在动情期表达显著高于其他3期(P0.05);ATF4蛋白主要在腺上皮细胞和腔上皮细胞胞质表达,其表达规律与RT-PCR的结果基本一致。结论 ATF4在小鼠动情周期子宫内膜中呈动态表达,提示ATF4可能参与了动情周期子宫内膜变化的调控。     

小鼠实验性肺纤维化上皮细胞-间充质细胞转变及骨桥蛋白的表达

目的从形态学上证实小鼠实验性肺纤维化上皮细胞-间充质细胞转变(EMT)及骨桥蛋白(OPN)的表达变化。方法将60只健康雄性C57BL/6小鼠随机分为生理盐水对照组和博来霉素(BLM)模型组,采用口咽抽吸法经咽部注入气管50μl博来霉素溶液,对照组注入50μl生理盐水。分别在造模后第3d、7d、14d、21d及28d 5个时间点处死对照组及模型组小鼠。取小鼠左肺制备石蜡切片,行苏木素-伊红(HE)染色及天狼猩红染色观察肺组织形态变化及胶原增生情况;行表面活性物质相关蛋白-C(SP-C)、上皮型钙黏蛋白(E-Cadherin)、波形蛋白(vimentin)、成纤维细胞特异蛋白1(FSP1)和骨桥蛋白(OPN)免疫组织化学染色,观察肺纤维化上皮细胞-间充质细胞转变(EMT)形态与OPN表达,利用图像分析软件Image-Pro Plus 6.0(IPP6.0)测量切片积分吸光度(IA),并进行统计分析。结果HE及天狼猩红染色显示出小鼠肺组织正常结构和纤维化形态变化,并且模型组小鼠肺组织的胶原生成量随时间推移而增加(P<0.05)。免疫组织化学结果显示,模型组小鼠肺OPN表达量随病程进展持续升高,在纤维化期达到峰值,而对照组表达量很少(P<0.05);对照组SP-C在Ⅱ型肺泡细胞恒定表达,模型组SP-C持续增加,且胞体增大;对照组E-Cadherin表达量多,而模型组在肺实变部位几乎不表达,但肺实变部位产生波形蛋白及FSP1,这些蛋白提示肺纤维化存在EMT。结论BLM诱导性肺纤维化中存在EMT,且与显著增加的OPN有潜在关系。     

妊娠早期小鼠卵巢、输卵管及子宫内诱导型一氧化氮合酶的分布

目的　了解胚泡着床前后妊娠昆明系小鼠卵巢、输卵管及子宫内诱导型一氧化氮合酶 (inducible ni-tric oxide synthase,i NOS)的分布。　方法　免疫细胞化学 L SAB法。　结果　妊娠 2～ 5 d小鼠的卵巢内 ,黄体细胞上有 i NOS的阳性表达 ;输卵管粘膜上有 i NOS的分布 ,肌层则为阴性 ;妊娠 2、3d的小鼠子宫内 ,阳性标记主要出现在子宫内膜上皮以及子宫内膜中的子宫腺上皮 ,内膜基质细胞为阴性 ;妊娠 4d的子宫内 ,子宫腺及蜕膜部分均有 i-NOS的分布 ,妊娠 5 d时 ,在小鼠胚泡的表面也检测到了 i NOS的存在。　结论　小鼠胚泡着床前后 ,在其卵巢、输卵管及子宫内均有 i NOS的存在 ,提示 i NOS在小鼠胚胎早期发育及着床过程中起作用。     

槲皮素对小鼠动情周期及免疫功能的调节作用

目的:研究槲皮素对小鼠动情周期的影响及其免疫效应,并探讨其作用机制。方法:对小鼠连续7天灌胃给予浓度为2.5 mg/ml的槲皮素药液0.5 ml/(天.只),通过阴道脱落细胞学检查法判断未经产雌鼠所处的动情周期,并利用MTT法检测脾淋巴细胞转化率、α-醋酸萘酯酶法检测T淋巴细胞数量、碳廓清实验检测吞噬细胞活性、定量溶血分光光度法检测IgM抗体生成量。结果:槲皮素显著延长了小鼠的动情周期,并可以明显增加淋巴细胞转化率和T淋巴细胞数目、增强小鼠巨噬细胞的吞噬活性和IgM抗体的生成。结论:槲皮素对小鼠子宫内膜的周期性变化造成了影响,对小鼠机体起到了免疫增强作用。     

小鼠动情周期内雌孕激素波动与皮层及海马δ-亚基的突触外γ-氨基丁酸A型受体表达的相关性

目的探讨雌性小鼠动情周期不同阶段血清孕酮浓度与大脑皮质和海马中含δ-亚基的突触外γ-氨基丁酸A型受体(δGABA_ARs)表达的相关性。方法动情间期和动情期成熟卵巢周期雌性小鼠(4～6周,n=12)血清雌激素和孕酮浓度采用ELISA试剂盒检测;动情间期和动情期大脑皮层和海马中δGABA_ARs表达采用免疫组织化学方法检测。结果在动情间期,皮层和海马δGABA_ARs表达明显高于动情期,血清孕酮的浓度与δGABA_ARs的积分吸光度值成线性关系。结论动情期不同阶段孕酮的生理波动会影响δGABA_ARs在海马及皮层等特定脑区的表达,而海马神经元δGABA_ARs表达的差异会影响脑区神经元的兴奋度,从而可能影响雌性动物的行为特性。     

骨桥蛋白在子宫内膜癌及子宫颈癌组织中的表达

目的：探讨骨桥蛋白(OPN)在子宫内膜癌及子宫颈癌组织中的表达及意义。 方法: 采用免疫组织化学SP法测定OPN在30例子宫内膜癌、20例子宫颈癌及30例正常对照组织的表达，并评价其与临床病理特征的关系。 结果: OPN在子宫内膜癌的阳性表达率为70%,显著高于正常增殖期子宫内膜（10%）及分泌期子宫内膜（50%），P<0.01; OPN在子宫颈癌的阳性表达率（55%）显著高于正常宫颈上皮(10%),P<0.01；而宫颈磷癌及宫颈腺癌组织的阳性表达率分别为53.3%及60.0%, 两者之间无统计学差异, P>0.05。OPN的阳性表达率随临床病理分期的增加及细胞分化程度降低而显著升高。 结论: OPN在子宫内膜癌及子宫颈癌组织均有显著高表达,其表达与肿瘤分期及分级密切相关。     

子宫颈鳞癌中骨桥蛋白的表达及其临床意义

目的观察骨桥蛋白(osteopontin,OPN)在子宫颈鳞癌中的表达及其临床意义,评估OPN是否可以作为子宫颈鳞癌的诊断和预后标记。方法采用免疫组化SP法对88例子宫颈鳞癌及18例正常子宫颈组织中OPN进行检测,并复习相关文献。结果免疫组化染色结果示OPN在子宫颈鳞癌的阳性率为59.09%(52/88),在18例正常子宫颈组织均阴性。子宫颈鳞癌组OPN阳性表达与子宫颈鳞癌的淋巴结转移(P=0.030)和无病生存时间有关(P=0.029),与患者年龄、子宫颈鳞癌浸润的深度、FIGO分期(Ⅰ、Ⅱ期)及病理分级无关(P>0.05)。结论 OPN在子宫颈鳞癌的转移、发展过程中扮演着重要角色,在治疗过程中对其进行检测有助于对子宫颈鳞癌患者的预后进行评估。     

骨桥蛋白对小鼠体外受精及早期胚胎发育的影响

目的 探讨骨桥蛋白（OPN）对小鼠体外受精及早期胚胎发育的影响。方法 120只昆明雌鼠和30只雄鼠,采用体外受精、胚胎培养方法,将小鼠精子、卵子、原核期胚胎和2-细胞期胚胎分别用不同浓度的OPN抗体预处理,观察小鼠受精、卵裂以及早期胚胎发育情况。结果 不同浓度的OPN抗体分别预处理精子和卵子后,与对照组比较,受精率显著降低(P＜0.01)。OPN抗体预处理原核期胚胎后,0.01mg/L OPN抗体组的卵裂率较对照组低,但差异无显著性 (P =0.052),1.00mg/L OPN抗体组的卵裂率低于0.01mg/L 组（P ＜0.01）及0.10mg/L组（P ＜0.05）。不同浓度的OPN抗体预处理2-细胞胚胎后可抑制胚胎发育。1.00mg/L OPN抗体组的4-细胞率和8-细胞率与0.10mg/L OPN抗体组相比差异无显著性(P ＞0.05),但前者的囊胚形成率显著低于后者（P ＜0.01）。1.00mg/L OPN抗体组的4-细胞率、8-细胞率以及囊胚形成率均显著低于0.01mg/L OPN抗体组（P ＜0.01）。结论 OPN可促进小鼠的受精和早期胚胎发育。     

Follicle stimulating hormone receptor protein is expressed in ovine uterus during the estrous cycle and utero-placenta during early pregnancy: An immunohistochemical study

Follicle stimulating hormone (FSH) is a well characterized gonadotropin that controls primarily development and functions of ovarian follicles in mammalian species. FSH binds to a specific G protein-coupled receptor (FSHR) belonging to the glycoprotein hormone receptor family that plays an essential role in reproduction. Although the primary location of FSHR is in the gonads (mainly in ovarian follicles), FSHR protein and/or mRNA have also been detected in extragonadal female reproductive tissues including embryo, placenta, endometrium, cervix, ovarian cancer tissues, and/or endometriotic lesions in several species. To determine the pattern of FSHR expression in the uterus and placenta, uterine tissues were collected at the early, mid- and/or late luteal phases of the estrous cycle from non-treated or FSH-treated ewes, and utero-placental tissues were collected during early pregnancy followed by immunohistochemistry and image generation. FSHR was immunolocalized to several uterine and utero-placental compartments including luminal epithelium, endometrial glands and surrounding stroma, myometrium, and endothelium and vascular smooth muscle cells in endometrium, myometrium and mesometrium. Intensity of staining and distribution of FSHR in selected compartments differed and seems to depend on the stage of the estrous cycle or pregnancy, and FSH-treatment. These novel data demonstrate differential expression of FSHR protein indicating that FSH plays a specific role in regulation of uterine and utero-placenta functions in sheep.     

Sexual preference in female rats during estrous cycle, pregnancy and lactation.

Choice of a male or female social contact was studied in intact female rats in a runway-choice apparatus during estrous cycle, pregnancy, and lactation. The male was chosen significantly more often during proestrus/estrus than during the diestrous days of the cycle. During pregnancy this preference in choice declined only to reappear gradually during the lactation period. The shifts in the level of motivation to seek out the male support previous studies and point to the significance of estrogen in producing the preference for the male.     

动情期和动情间期小鼠输卵管中氧化应激和抗氧化基因表达差异

目的观察昆明(Kunming,KM)小鼠输卵管中氧化应激和抗氧化基因的表达是否随动情周期而改变。方法分别取动情期和动情间期小鼠输卵管进行RNA提取,用PCR芯片检测氧化应激和抗氧化基因表达的差别,然后用Real Time PCR对差异表达基因进一步验证。结果与动情间期比较,动情期小鼠输卵管中NADPH氧化酶1(NADPH oxidase1,NOX1)、谷胱甘肽过氧化物酶2(glutathion peroxidase 2,GPX2)、超氧化物歧化酶3(superoxide dismutase 3,SOD3)、一氧化氮合酶2(nitric oxide synthase 2,NOS2)等4个基因表达上调。结论动情期小鼠输卵管中NOX1、GPX2、SOD3和NOS2等4个基因表达上调,提示其可能参与维持动情期小鼠输卵管腔内适度的活性氧自由基(reactive oxygen species,ROS)和一氧化氮(nitric oxide,NO)水平,为卵子受精和早胚发育提供必要第二信号分子。     

Changes in histochemical distribution of cell surface heparan sulfate proteoglycan in mouse uterus during the estrous cycle and early pregnancy

We have investigated the changes in immunolocalization of a cell surface heparan sulfate proteoglycan (HSPG) in the mouse uterus during the estrous cycle and at the time of implantation in early pregnancy. A monoclonal antibody prepared against syndecan, a cell surface HSPG from mouse mammary epithelium (gift of Dr. M. Bernfield), was reacted with unfixed and fixed frozen sections of uteri from normally cycling, 3.5 and 4.5 days pregnant, and estradiol-treated immature and ovariectomized mature mice. A polyclonal antibody prepared against basal lamina HSPG from Engelbreth-Holm-Swarm (EHS) tumor cells (gift of Dr. John Hassell) was used as a positive control. The latter showed no variation during the estrous cycle or early pregnancy. Localization of syndecan in uterine epithelium changed from basolateral to predominantly basal as the cycle progressed from metestrus toward estrus. A similar pattern was seen in immature and ovariectomized mature females that had received estradiol injections. With the onset of pregnancy, the basolateral localization became progressively less intense from 3.5 days through 4.5 days of pregnancy. Thus, cell surface HSPG distribution is modulated by hormonally dependent changes in cycling and pregnant mice, supporting previous suggestions that early pregnancy in mice is accompanied by a turnover and rearrangement of uterine epithelial cell surface.© Willey-Liss, Inc.     

Changes in bradykinin and bradykinin B2-receptor during estrous cycle of mouse

The aim of the study was to investigate changes in the abundance of bradykinin and bradykinin B2-receptor in the ovary of mice during its estrous cycle. Changes in the abundance of bradykinin were correlated with changes in bradykinin B₂-receptor in order to determine the functional significance of this peptide for follicular development, ovulation and luteinization. Bradykinin immunoreactivity was mainly observed in the granulosa cells of antral follicles, especially around the oocytes and lining the antral cavity during proestrus and estrus phases of the cycle. Recently formed corpora lutea showed mild immunoreactivity for both bradykinin and bradykinin B₂-receptor. During diestrus 1 and diestrus 2, bradykinin and bradykinin B₂-receptor immunostaining was mainly found in the corpora lutea and mildly in the antral follicles. Immunoblot analysis for bradykinin and bradykinin B₂-receptor attained a peak during late evening in proestrus, which may be the time of the LH surge. Thereafter bradykinin and bradykinin B₂-receptor declined sharply during the estrus phase. When the concentration of bradykinin was correlated with bradykinin B₂-receptor throughout the estrous cycle, they showed strong positive correlation. Thus, this study indicates that the levels of bradykinin and bradykinin B₂-receptor both simultaneously regulate estrous cycle and are important components for the reproductive process.     

Expression of repulsive guidance molecule b (RGMb) in the uterus and ovary during the estrous cycle in rats

Repulsive guidance molecule b (RGMb; a.k.a. Dragon), initially identified in the embryonic dorsal root ganglion, is the first member of the RGM family shown to enhance bone morphogenetic protein (BMP) signaling by acting as a BMP co-receptor. BMP signaling has been demonstrated to play an important role in the reproductive organs. Our previous study found that RGMb was expressed in the reproductive axis, but whether RGMb expression in reproductive organs changes across the estrous cycle remains unknown. Here, we show in the rat that RGMb mRNA expression in the uterus was significantly higher during metesterus and diestrus than during proestrus and estrus. Western blotting indicated that RGMb protein was significantly lower during estrus compared with the other three stages. Immunohistochemistry revealed that RGMb protein was mainly localized to the uterine luminal and glandular epithelial cells of the endometrium. RGMb mRNA and protein in the ovary remained unchanged during the estrous cycle. RGMb protein was expressed in the oocytes of all follicles. Weak staining for RGMb protein was also found in corpora lutea. RGMb was not detected in granulosa cells and stromal cells. Taken together, RGMb expression in the uterus and ovary across the estrus cycle demonstrate that RGMb may be involved in the regulation of uterine function, follicular development as well as luteal activity.     

Localization and distribution of platelet activating factor receptors in the mouse ovary and oviduct during the estrous cycle and early pregnancy

PROBLEM: This study determined platelet activating factor (PAF) binding to ovaries and oviduct tissues to identify the location of PAF tissue interaction. METHOD: Mouse ovaries and oviducts taken during the estrous cycle and days 1, 4 and 7 post-conception (pc) were analyzed by [3H]PAF binding using frozen sections and autoradiography. RESULTS: For the outer epithelium of estrous and metestrous ovaries there was no significant difference in PAF binding (P>0.05); however, both stages were significantly different from proestrous (P<0.001). Ovarian stroma PAF binding was significantly higher (P<0.001) at estrous than metestrous, with no PAF binding at proestrous. Binding of PAF to estrous ovarian follicles was significantly greater (P<0.001) than at proestrous and metestrous. Estrous oviduct binding of PAF was significantly increased for stroma, inner epithelium and lumen (P<0.001 in all cases). At days 1 and 7 pc, all ovarian tissues had the greatest PAF binding with day 4 pc failing to bind PAF except for a significant decrease in corpora luteal binding (P<0.001). Oviduct binding of PAF was greatest at day 1 pc. CONCLUSION: Ovarian and oviduct PAF receptor expression corresponds to peak embryo PAF synthesis and establishes the basis for PAF PAF receptor mediated early pregnancy signaling system.     

Neurokinin A in the hypothalamus and anterior pituitary during the estrous cycle in the golden hamster

The fluctuations in the neurokinin A concentrations in the hypothalamus and anterior pituitary of female golden hamsters were studied in the different stages of the estrous cycle, and they were correlated with the changes in serum estradiol levels. Neurokinin A levels in the hypothalamus were lowest at day 4 of the cycle (proestrus), when serum estradiol levels were highest. Neurokinin A levels in the hypothalamus of hamsters in day 1 (estrus), day 2 (diestrus I), and day 3 (diestrus II) were not significantly different from each other. In the anterior pituitary, the highest neurokinin A concentrations were found during day 1 of the cycle (estrus), the levels in day 2 (diestrus I), day 3 (diestrus II), and day 4 (proestrus) were significantly lower than at estrus, again showing the lowest levels during proestrus, when estradiol levels were maximal. These results show that neurokinin A levels in the hypothalamus and anterior pituitary of the female hamster undergo significant changes during the estrous cycle.     

Effect of progesterone on the estrous activity cycle of the rat

Two, 3 or 4 Silastic capsules of progesterone were implanted sc for 24–35 days, removed for a 12–14 day interval and re-implanted into the same adult female rats from which they were taken. A control group received cholesterol capsules. For approximately 4–10 days, Silastic capsules of progesterone suppressed estrous activity cycles. Thereafter, amplitude of running increased progressively and definite periodicities re-emerged. Following a 12 day recovery period, the same capsules were re-implanted and activity was again suppressed. During the first implantation, the number of estrous cycles with greater than 4-day periods significantly increased. Progesterone was also found to delay the daily onset of activity in relation to the light/dark cycle (phase angle difference). Phase angle difference prior to the first implantation was negatively correlated with the rate of recovery. The data support the conclusion that progesterone affects parameters of estrous activity by antagonizing the actions of estrogen and suggest that animals may make internal adjustments to high titers of progesterone.     

p16INK4A在动情周期和早孕期小鼠子宫内膜的表达

目的 检测肿瘤抑制基因p16INK4A在小鼠动情周期和早孕期子宫内膜中的表达,初步探讨p16^INK4A在小鼠胚胎植入过程中的生物学作用。方法 运用反转录-聚合酶链反应(RT-PCR)技术检测动情周期和早孕期小鼠子宫内膜p16^INK4AmRNA的表达水平;用免疫组织化学和Western blotting方法检测p16INK4A蛋白的表达。结果 RT-PCR显示,早孕期比动情周期p16^INK4AmRNA的表达有显著增强。动情周期中动情期比其他3期表达明显增强;早孕期中,随妊娠天数的增加,p16^INK4AmRNA的表达也逐渐增强,孕5d达峰值,之后又逐渐减弱。免疫组织化学和Western blotting的结果也都显示,p16^INK4A蛋白在子宫内膜的表达及规律与RT-PCR的结果一致。结论 p16^INK4A在小鼠的胚泡植入过程中起重要作用。