中药催情助孕液治疗奶牛卵巢性不孕症

在影响奶牛繁殖的诸多因素中,奶牛卵巢性不孕症比较常见,而且是影响奶牛正常繁殖和发展的主要因素.笔者应用中药制剂催情助孕液治疗卵巢静止、卵巢囊肿和持久黄体等卵巢性不孕症,取得了比较满意的效果.     

传染性支气管炎病毒感染对蛋雏鸡输卵管雌激素受体及孕激素受体表达的影响

以不同传染性支气管炎病毒（肾型IBV-河北分离株与呼吸型IBV-M41）分别感染蛋雏鸡,通过测定输卵管蛋白分泌部及子宫雌激素受体（Estrogen receptor,ER）和孕激素受体（Progesterone receptor,PR）的表达水平,来探讨IBV早期感染（10d）对抑制蛋鸡输卵管发育的作用机制。结果显示,IBV感染降低子宫和蛋白分泌部ER mR-NA、PR mRNA的表达水平,即致子宫ER mRNA和PR mRNA的表达低于检测水平;蛋白分泌部ER mRNA和PR mRNA的表达水平显著降低（P〈0.01）,且M41的作用幅度较肾型IBV更为显著（P〈0.01）。结果表明,IBV可通过降低ER和PR的表达水平,抑制输卵管对雌激素的应答能力而影响输卵管的发育。     

犬阴道肿瘤雌孕激素受体的表达

将吉林大学小动物医院2006年6月至2008年2月收治的35例犬阴道肿瘤,根据临床特征及病理学变化分为传染性性病肿瘤、阴道增生和腺泡状软组织肉瘤3种,采用免疫组化SP法检测雌二醇受体α(ER-α)和孕酮受体(PR)在肿瘤中的表达。结果表明,犬传染性性病肿瘤(Canine transmissible venereal tumor,CTVT)组织雌孕激素受体表达缺失。阴道增生雌孕激素受体的表达,发情前期上皮细胞、平滑肌细胞及内皮细胞与正常阴道组织相比阳性表达率相似,并未随发情前期血浆雌二醇水平的升高而增加。而腺泡状软组织肉瘤(Vagin alalveolar soft part sarcoma,VASPS)雌二醇受体表达为阳性,表明犬阴道腺泡状软组织肉瘤的生成、发展可能与生殖激素有关,具有激素依赖性。     

M41型IBV对输卵管中雌激素、孕激素受体及胰岛素生长因子-Ⅰ表达的影响

本试验通过相对定量RT-PCR检测接种M41型鸡传染性支气管炎病毒(infectious bronchitis virus,IBV)对蛋鸡输卵管各部分(蛋白部、峡部、伞部和子宫)雌激素受体(ER)、孕激素受体(PR)及胰岛素生长因子-Ⅰ(IGF-Ⅰ)mRNA表达的影响,研究IBV早期感染(10 d)抑制蛋鸡输卵管发育的作用机制。结果显示,输卵管各部均有ER、PR和IGF-Ⅰ mRNA的表达,染毒组ER、PR mRNA的表达量较正常对照组显著减少(P＜0.05),而IGF-Ⅰ的表达量较正常对照组显著增加(P＜0.05)。可见IBV可通过降低ER和 PR mRNA的表达水平,抑制输卵管对雌激素的应答能力而影响输卵管的发育;而IGF-Ⅰ可能是以自分泌、旁分泌方式在输卵管局部起作用,是输卵管生长的调节因子。     

黄芩对ZEA暴露雌性小鼠抗氧化能力、性激素水平和ERα mRNA表达的影响

【目的】试验旨在研究黄芩(SBG)对玉米赤霉烯酮(ZEA)暴露幼龄雌性小鼠抗氧化能力、性激素水平及雌性生殖器官中雌激素受体α(ERα)和孕酮受体(PR)基因表达的影响。【方法】试验选取50只雌性小鼠,预饲1周后随机分为5组,每组10只。对照组(CK)灌胃10 mL/kg玉米油、ZEA组灌胃20 mg/kg ZEA、ZEA+SBG10组灌胃20 mg/kg ZEA+10 g/kg SBG、ZEA+SBG20组灌胃20 mg/kg ZEA+20 g/kg SBG、ZEA+SBG30组灌胃20 mg/kg ZEA+30 g/kg SBG。所有试验小鼠均饲喂基础饲粮,正试期3周。正试期间每日记录小鼠体重;分别在试验的第1、2和3周末晨饲前采集小鼠尾静脉血样,检测血清中抗氧化相关指标和性激素含量;在试验第3周末,采集卵巢和子宫,检测相关类固醇激素受体基因mRNA的表达。【结果】与CK组相比,ZEA暴露2周以上可引起雌性小鼠体重、血清谷胱甘肽过氧化物酶(GSH-Px)活性、谷胱甘肽(GSH)和雌二醇(E₂)含量均显著降低(P<0.05);卵巢刺激素(FSH)水平显著增加...     

慢性砷暴露对雌鼠乳腺组织ER mRNA和PR mRNA表达的影响

建立慢性砷暴露小鼠动物模型,通过实时定量PCR方法检测慢性砷暴露小鼠乳腺组织中ER、PR mRNA表达量。结果显示,(1)对照组mRNA表达量ER为0.0025±0.0011、PR为0.0190±0.0150;(2)0.05mg/L组ER为0.0085±0.0080、PR为0.0470±0.0180;(3)0.1mg/L组ER为0.0027±0.0003、PR为0.0210±0.0200;(4)0.2mg/L组ER为0.0037±0.0025、PR为0.0390±0.0130;(5)0.4mg/L组ER为0.0029±0.0018、PR为0.0072±0.0031。与对照组比较,各组砷暴露小鼠ER表达均升高,其中0.05mg/L组ER表达差异显著(P0.05);除0.4mg/L组表达降低外,其他各暴露组PR表达也升高,0.05mg/L组PR表达差异极显著(P0.01);0.2mg/L组的PR表达差异显著(P0.05)。结果表明,慢性砷暴露小鼠乳腺组织中ER、PR表达量较对照组发生改变,可能作为一种环境内分泌干扰物发挥雌激素效应进而产生毒性作用。     

“益母散”对去卵巢小鼠发情和雌激素受体表达的影响

旨在研究纯中药制剂益母散是否对小鼠子宫有直接作用并影响小鼠的发情和雌激素受体的表达。以性成熟雌性小鼠为研究对象,通过手术去除小鼠卵巢,用益母散进行灌胃试验,试验期间每天进行阴道涂片,检测小鼠的发情情况,试验结束后取出子宫,测子宫质量,同时对子宫组织HE染色观察子宫的组织形态,通过PCR检测子宫雌激素受体表达,并测定发情期血清雌激素水平。结果表明,益母散可明显抑制去卵巢小鼠子宫的萎缩(P0.01),并能促进去卵巢小鼠子宫发育、维持子宫正常组织形态、促进雌激素受体ERαmRNA表达,使去卵巢小鼠出现发情表现。这表明益母散具有一定的类雌激素样作用,可通过上调雌激素受体表达,使去卵巢小鼠在内源性雌激素水平较低的情况下出现发情表现。     

出生后发育时期济宁青山羊乳腺中ERα和PR的分布及其mRNA表达

本研究旨在探讨济宁青山羊出生后发育过程中乳腺组织内雌激素受体α(ERα)和孕激素受体(PR)免疫阳性细胞的分布及平均光密度和mRNA分子水平的表达规律.应用SP免疫组化法和图像分析方法检测ERα和PR在济宁青山羊乳腺内的分布;以GAPDH为内参基因,qRT-PCR方法检测ERα和PR mRNA的相对表达量.结果表明,ERα和PR阳性染色主要见于乳腺组织内的血管内皮细胞、基质细胞和腺管上皮细胞的细胞核中,偶见胞浆阳性着色,180日龄在腺泡上皮细胞亦可见PR阳性表达;ERα和PR平均光密度自出生～性成熟期呈现逐渐上升的趋势,且性成熟期显著高于出生当天和初情期(P＜0.05),但与180日龄相比无显著差异(P＞0.05).qRT-PCR结果显示,ERα mRNA的表达量自出生～性成熟期呈现逐渐上升趋势,性成熟期最高,极显著高于出生当天和初情期(P＜0.01),180日龄略有下降.PR mRNA表达量自出生当天～性成熟期较低且组间差异不显著(P＞0.05),180日龄的表达量最高且极显著高于其他的3组(P＜0.01).济宁青山羊出生后发育过程中,ERα对腺管伸长和上皮分化以及PR对腺管分支和腺泡发育起促进作用.     

胰岛素对体外培养牛肝细胞胰岛素受体mRNA水平的影响

在新生牛单层肝细胞培养液中添加胰岛素,其浓度分别为0μmol/L、5μmol/L、10μmol/L、20μmol/L、50μmol/L和100μmol/L,处理12h,应用半定量RT-PCR方法研究胰岛素对体外培养新生牛单层肝细胞胰岛素受体(IR)mRNA水平的影响.结果表明随胰岛素浓度的升高,肝细胞IRmRNA丰度呈下降趋势,指示肝细胞内IRrnRNA丰度下降以调节胰岛素浓度.     

不同营养水平限制饲养对妊娠期蒙古绵羊生长激素受体(GHR)mRNA表达水平的影响

为了阐明不同营养水平限制饲养妊娠期母羊后,在不同生长阶段对蒙古绵羊生长激素受体(GHR)mRNA表达水平的影响,研究根据GenBank中绵羊的GHR序列,设计1对预计扩增产物为395 bp的引物,采用RT-PCR技术检测了蒙古绵羊在不同营养水平条件下肝脏和背最长肌中GHR mRNA表达量的变化。结果表明:在限制采食阶段,低营养水平组即营养限制组(RG1组)和阈值组(RG2组)肝脏的GHR mRNA表达水平低于正常营养的对照组(CG组),而背最长肌的GHR mRNA表达水平高于对照组,说明限制营养对绵羊GHR水平的影响具有组织特异性。在补偿生长结束阶段,RG1组和RG2组绵羊的肝脏和背最长肌中GHR mRNA表达水平均恢复到对照组水平。     

Bovine uterine, cervical and ovarian androgen receptor concentrations. Correlation with estrogen and progesterone receptor concentrations.

Bovine cytosol androgen receptor (ARC) concentrations were examined simultaneously in various regions of the uterus and in ovarian tissues of cows, and were related to cytosol estrogen (ERC) and progesterone receptor (PRC) concentrations and circulating steroid levels. ERC concentrations were 3-7-fold and PRC concentrations 13-29-fold those of ARC in bovine endometrial and myometrial tissues. When serum progesterone levels were low, both endometrial and myometrial ARC, endometrial ERC, and endometrial and myometrial PRC concentrations were higher (p < 0.05) than those observed during higher progesterone concentrations. Because serum 5 alpha-dihydrotestosterone (5 alpha-DHT) concentrations were higher during the luteal phase, it is possible that ARC was down-regulated by this natural ligand at this phase of the cycle. There were no differences between uterine horns in endometrial or myometrial ARC concentrations. Bovine cervical and ovarian stromal tissue also contained ARC, and the concentrations were about the same as in the endometrium and the myometrium. The relative binding affinities (RBAs) of some steroid hormones towards ARC in vitro were: the synthetic compound R1881 (146%), 5 alpha-dihydrotestosterone (100%), testosterone (75%) while estradiol-17 beta, progesterone and dexamethasone had lower RBAs (2, < 1, < 1% respectively). Cytosol androgen receptor concentrations correlated significantly with cytosol progesterone (PRC) and estrogen receptor (ERC) concentrations, both in the endometrium and myometrium. These data show that androgens, such as 5 alpha-DHT, may participate the endocrine regulation of bovine reproductive tissues.     

Expression of estrogen receptor 1 and progesterone receptor in primary goat mammary epithelial cells

The exact role and sensitivity of cells to estrogen and progesterone mediated through the steroid receptors during lactation is not known. Expression of estrogen receptor 1 (ESR1) and progesterone receptor (PGR) was quantified in mammary tissue‐derived primary goat mammary epithelial cells (pgMECs) to determine the influence of donor tissue physiology (lactating and juvenile) and cell culture growth conditions (basal and lactogenic) on ESR1 and PGR expression in the derived cells. Relative messenger RNA (mRNA) levels for both receptors were the highest in cell lines derived from mammary tissue of juvenile goats. Maintaining pgMECs in lactogenic conditions resulted in up‐regulation of ESR1 (1.36‐ to 12.35‐fold) and in down‐regulation of PGR (‐2.53‐ to ‐3.62‐fold), compared to basal conditions. Based on Western blotting analysis we suggest that the differences in mRNA expression are translated to the protein level. We suggest that differential expression in lactating conditions is correlated with terminal differentiation of the pgMECs. Double immunostainings showed that estrogen receptor alpha (ER‐α) positive cells do not exclusively belong to the luminal lineage and that ER‐α and PGR can be expressed individually or co‐expressed in the pgMECs. The derived primary cultures/lines in early passages are hormone‐responsive and represent a useful surrogate for mammary tissue in research experiments.     

Immunohistochemical characterization of estrogen and progesterone receptors in lymphoma of horses

Abstract: Immunohistochemical techniques were used to examine 29 cases of equine lymphoma for estrogen receptor (ER) and progesterone receptor (PR) expression. The lymphomas examined included T-cell-rich large B-cell lymphomas, B-cell neoplasms, and T-cell lymphomas. The individual cases were also classified according to the anatomic location of the tumors. One normal equine lymph node was also examined for ER and PR expression. All of the cases of equine lymphoma and the normal lymph node were negative for ER. A total of 16/29 (55%) PR-positive lymphomas were identified. Seven of the 12 (58%) T-cell-rich large B-cell lymphomas were positive, 7/11 (64%) B-cell tumors were positive, and 2/6 (33%) T-cell neoplasms were positive. Anatomically, 6/9 (66%) subcutaneous lymphomas were PR positive, 3/5 (60%) intrathoracic lymphomas were positive, 1/4 (25%) intra-abdominal lymphomas were positive, 2/5 (40%) intra-abdominal/intrathoracic lymphomas were positive, 1/2 (50%) upper airway lymphomas were positive, and 3/3 (100%) splenic lymphomas were positive. One case involving abdominal and thoracic tumors and leukemia was negative for PR expression. The normal lymph node contained a low percentage (1.9%) of PR-positive lymphocytes. The presence of PR in neoplastic equine lymphoid tissue indicates that these tumors may be responsive to serum progesterone. Also, identification of PR-positive cells in the normal lymph node suggests that PR may be constitu-tively expressed in normal equine lymphocytes. Further studies are needed to quantify PR levels in normal and malignant equine lymphoid tissue and to determine the usefulness of either progestin or antiprogestin drugs in the management of equine lymphoma.     

大鼠不同时期乳腺组织中雌激素孕激素及催乳素的变化规律

为探讨雌激素(E2)、孕激素(P)及催乳素(Prl)在大鼠不同时期乳腺组织中的变化规律,选雌性SD大鼠42只,分别为处女鼠、妊娠鼠(6d、12d、18 d)、泌乳鼠(6d、12d、18 d),每个时期6只.在指定的时间点处死大鼠取乳腺组织.用放免(RIA)组织匀浆中E2、P及Prl水平进行定量研究,用统计学方法进行处理,观察这三种激素的变化规律.结果显示E2从处女期到整个妊娠期,其呈现下降趋势.而分娩后E2水平急剧升高,后又有所下降,但维持在较高水平;在乳腺发育不同时期,P在乳腺组织中整体水平一直呈现下降趋势;Prl水平以处女期最高;随着妊娠进行逐渐降低,整个乳腺发育过程中Prl整体上几乎一直呈下降趋势.表明E2、P及Prl在促进乳腺发育及维持和启动泌乳方面发挥了重要的作用.     

Expression and localization of estrogen receptors α and β mRNA in medullary bone of laying hens

The aim of this study was to observe the expression and localization of estrogen receptor (ER) α and ER β mRNA in the medullary bone of laying hens. First, medullary bone, liver, kidney, and shell gland of the oviduct tissues were dissected from laying hens. Then, the total cellular RNA was isolated from each tissue specimen, and the ER α and ER β mRNA expression was observed using semiquantitative RT‐PCR. Second, the localization of ER α mRNA in the medullary bone was detected with in situ hybridization using digoxigenin‐11‐UTP‐labeled cRNA probes. As a result, the expression of ER α mRNA was higher than that of ER β mRNA in the medullary bone, liver, and shell gland of the oviduct from laying hens. In the kidney, ER α mRNA expression was lower than that of ER β mRNA. The expression pattern of ER α and ER β mRNA of the medullary bone was similar to that of the shell gland of the oviduct. Moreover, ER α mRNA was intensively expressed in osteoblasts on the medullary bone surface and bone marrow stromal cells but was not expressed in osteoclasts. These results suggest that in medullary bone, estrogen action may be regulated not by ER β but by ER α.     

蒙古马不同组织器官TLR1、TLR2、TLR4和TLR6 mRNA转录水平研究

根据GenBank中马Toll样受体基因序列设计特异性引物,建立检测马Toll样受体（TLRs）mRNA转录水平的SYBR GreenⅠ实时荧光定量PCR方法,检测TLR4、TLR2、TLR1和TLR6在蒙古马不同组织器官中的转录水平。4种TLRs在心脏、肝脏、脾脏、肺脏、肾脏、胃、十二指肠、空肠、盲肠和骨髓中均有转录。其中,TLR4mRNA除在空肠和肝脏外,在其他组织器官中的表达水平均高于TLR2、TLR1、TLR6。免疫器官中,TLR4、TLR1mRNA在骨髓中表达量高于脾脏,而TLR2、TLR6mRNA在脾脏中表达量高于骨髓。各肠段,TLR4、TLR2、TLR1、TLR6mRNA表达水平之间在空肠的差异不是很大,而在十二指肠和盲肠中差异很大。结果表明,TLRs mRNA在马各组织器官转录水平差异较大,可能与其对病原体的识别和抵抗能力有关。     

子宫内膜炎患牛子宫内膜PGR和ESR-1 mRNA的表达特征

为探讨卵巢激素及其子宫内膜受体在奶牛子宫内膜炎中的作用,以产后6～10d健康及患急性化脓性子宫内膜炎的中国荷斯坦奶牛为研究对象,通过ELISA法检测子宫内膜中PGF2a,血清中孕酮和雌二醇浓度,SYBRGreenⅠ荧光定量PCR检测子宫内膜PGR和ESR-1mRNA的表达。结果显示,病牛子宫内膜PGF2α水平为（6.503±0.122）μg/L,血清孕酮为（0.816±0.281）μg/L,血清雌二醇为（35.462±2.523）μg/L,而对照组分别为（14.408±0.285）,（0.543±0.142）,（37.6±1.317）μg/L;子宫内膜炎患牛组PGF2a水平极显著低于对照组（P〈0.01）,孕酮含量显著高于对照组（P〈0.05）,PGF2a与孕酮水平呈显著负相关（r=0.893）,雌二醇含量组间差异不显著。PGR基因的mRNA水平极显著高于对照组（P〈0.01）,而ESR-1基因的mRNA水平低于对照组,且差异极显著（P〈0.01）。上述结果表明,孕酮浓度显著升高及PGR mRNA高表达,可能抑制子宫收缩;ESR-1mRNA低表达使子宫新陈代谢减慢,子宫内膜合成PGF2a功能下降,这些改变有可能促进了产后子宫内膜炎的发生发展。