抑制素亚基基因在哺乳类动物卵巢的表达与调控的研究进展

抑制素是由二硫键连接的α和β两个不同亚单位所构成的糖蛋白,它具有抑制卵泡刺激素（ＦＳＨ）的合成和分泌的作用。β亚单位借助二硫键尚可形成β亚单位二聚体－激活素,它则具有刺激ＦＳＨ分泌的作用。本文介绍了抑制素亚基ｃＤＮＡ的克隆及抑制素亚基基因在哺乳类卵巢表达与调控的研究进展,并指出在研究抑制基因表达时应予注意的问题。     

卵泡闭锁与其tPA，LH受体及抑制素 α，βA亚基基因表达的关系

用DNA 3′-末端标记、免疫组化和原位杂交方法,通过连续切片比较研究了相同卵泡颗粒细胞抑制素亚基和LH受体（LHR）与卵细胞tPA表达和卵泡闭锁的关系.实验结果表明（1） 卵泡闭锁伴随卵细胞tPA活性明显增加;（2） 颗粒细胞抑制素的产生调节卵细胞tPA活性的表达并与卵泡发育状态密切相关;（3） 卵泡闭锁时,颗粒细胞几乎不表达LHR和抑制素亚基.上述结果提示卵细胞的tPA在闭锁卵泡中可能参与卵细胞的自我瓦解和清除过程;颗粒细胞表达的抑制素可能是tPA mRNA翻译的一种抑制因子,如其表达受阻,可导致卵细胞tPA蛋白活性增加引起卵泡闭锁.     

卵泡闭锁与其tPA,LH受体及抑制素α,βA亚基基因表达的关系

用DNA 3′-末端标记、免疫组化和原位杂交方法,通过连续切片比较研究了相同卵泡颗粒细胞抑制素亚基和LH受体(LHR)与卵细胞tPA表达和卵泡闭锁的关系.实验结果表明:(1) 卵泡闭锁伴随卵细胞tPA活性明显增加;(2) 颗粒细胞抑制素的产生调节卵细胞tPA活性的表达并与卵泡发育状态密切相关;(3) 卵泡闭锁时,颗粒细胞几乎不表达LHR和抑制素亚基.上述结果提示:卵细胞的tPA在闭锁卵泡中可能参与卵细胞的自我瓦解和清除过程;颗粒细胞表达的抑制素可能是tPA mRNA翻译的一种抑制因子,如其表达受阻,可导致卵细胞tPA蛋白活性增加引起卵泡闭锁.     

人内皮生长抑制素基因克隆表达及其抑瘤作用

内皮生长抑制素（ｅｎｄｏｓｔａｔｉｎ是最近报道具有抑制肿瘤生长和铁蛋白质,为此,从正常人肝细胞株Ｌ０２中抽提总ＲＮＡ,以ＲＴ－ＰＣＲ法获得了人内皮生长抑制素编码序列,序列分析表明,内皮生长抑制素ＣＤＮＡ开放阅读框架全长５５５ｂｐ,此基因（ＧｅｎＢａｎｋ登录号为Ａ地８４０６０）,共编码１、８４个氨基酸残序列为５个碱基与文献报道不同,蛋白质序列中有３个氨基酸残基与文献报道不同,说明存在人种间差异。将人     

人血管抑制素在酿酒酵母中的分泌表达和活性鉴定

将酵母交配因子 (MF)α1信号肽编码序列和人血管抑制素 (hAGN)cDNA融合序列插入穿梭载体pYADE4 ,构建得到分泌型重组表达质粒pYADEMA18.转化酿酒酵母JG110 7后 ,用 2 %乙醇和2 %甘油联合诱导表达 .ELISA分析表明 ,在诱导 14h～ 30h期间 ,hAGN获得了表达并分泌至细胞外 .发酵上清液经 75 %饱和度硫酸铵沉淀、CM 5 2纤维素离子交换层析和Superdex 75凝胶过滤层析纯化 .SDS PAGE分析显示 ,表达产物重组人血管抑制素 (rhAGN)相对分子质量约 5 0kD ,电泳纯度达到 94 %.生物活性分析证明 ,rhAGN在 0 0 1mg L～ 3 0mg L浓度范围内能够抑制人真皮内皮细胞株HDMEC增殖 ,抑制作用随剂量的增加而增强 .     

抑制素的生理作用

抑制素是近年来提纯的一种主要由性腺分泌的糖蛋白激素,其主要作用是抑制腺垂体分泌泡刺激素、胎盘激素、胎盘激素的分泌、对生殖细胞的发育成熟及生殖活动都有一定的影响。     

抑制素基因的研究进展

抑制素是性腺分泌的一种糖蛋白激素,它具有抑制垂体促卵泡素合成和分泌的作用。本文介绍了抑制素α亚基基因（INHA）、抑制素βA亚基基因（INHBA）、抑制素βB亚基基因（INHBB）的克隆、结构、定位、多态性、表达、分子调节及其与繁殖性能和癌症的关系。绵羊INHA、INHBA和INHBB基因分别被定位到2q41→q43、4q26和2q31→q33。INHA、INHBA和INHBB基因对绵羊产羔数都有显著的影响。抑制素βB亚基基因突变的雌性小鼠有明显的发育和繁殖缺陷。INHA 基因与妇女卵巢早衰显著相关。 Abstract：Inhibins are gonadal glycoprotein hormones belonging to the transforming growth factor-βsuperfamily that act to suppress pituitary follicle-stimulating hormone synthesis and secretion. In this paper, we briefly introduced the cloning, structure, localization, polymorphism, expression, molecular regulation of inhibin-α(INHA), -βA (INHBA) and -βB (INHBB) subunit genes and their relationships with reproductive performance and cancer. The inhibin genes (INHA, INHBA and INHBB) had significant effect on litter size in sheep. The ovine INHA, INHBA and INHBB genes had been mapped to chromosomes 2q41→q43, 4q26 and 2q31→q33, respectively. The female mice carrying INHBB mutations suffered from distinct developmental and reproductive defects. The INHA gene was significantly associated with premature ovarian failure in women.     

内皮细胞抑制素的研究进展

内皮细胞抑制素是一种新发现的特异的血管内皮细胞增殖抑制剂,它可以有效地抑制内皮细胞生长,对肿瘤血管形成具有强烈的抑制作用,在抗实体瘤方面有显著的效果。本文就它的发现、结构、生物学活性、作用机理及研究进展进行综述。     

超声在诊断卵巢黄体破裂中的应用

目的探讨超声对卵巢黄体破裂的诊断价值。方法回顾性分析35例经超声诊断卵巢黄体破裂的声像特点及临床表现。结果35例均有不同程度的腹盆腔积液。24例手术治疗并病理证实,11例保守治疗,超声表现为盆腔积液,卵巢囊肿,附件区包块。结论超声对卵巢黄体破裂的早期快速诊断和制定治疗方案及愈后,有及其重要价值和意义。     

Differential Cellular Localization of Galectin-1 and Galectin-3 in the Regressing Corpus Luteum of Mice and Their Possible Contribution to Luteal Cell Elimination

Galectin-1 and galectin-3, β-galactoside–binding lectins, are predominantly expressed in the regressing corpus luteum (CL) of mouse ovary. This study revealed the expression patterns and cellular localizations of galectins during CL formation and regression by ISH and IHC. Galectin-1 mRNA expression temporarily increased in active CL, preceding the expression of progesterone degradation enzyme 20α-hydroxysteroid dehydrogenase (20α-HSD), which represents functional luteolysis. The expressions of both galectin-1 and galectin-3 remarkably increased in the structurally regressing CL, which vigorously expressed 20α-HSD and contained abundant apoptotic luteal cells. Ultrastructurally, galectin-1– and galectin-3–immunoreactive cells were identified as fibroblasts and infiltrating macrophages, respectively. In addition, some populations of luteal cells themselves expressed galectin-3 in regressing CL and formed unique demarcation membranes in the cytoplasm, showing a non-typical apoptotic feature. Ovary of adult mice with repeated estrus cycles contained CL of three different generations. Among them, the old CL formed during previous estrus cycles consisted of galectin-3–positive luteal cells. The galectin-3–positive old CL was resistant to apoptosis and seemed to be eliminated by a mechanism different from apoptosis. The stage- and cell-specific expression of galectin in CL suggests its differential contribution to luteolysis, and this expression may be mediated by major regulatory molecules of CL function, prolactin and/or prostaglandin F2α. (J Histochem Cytochem 58:741–749, 2010)     

Inhibin/activin subunits alpha, beta-A and beta-B are differentially expressed in normal human endometrium throughout the menstrual cycle

Inhibins are dimeric glycoproteins composed of an alpha () subunit and one of two possible beta (-) subunits (A or B). The aims of this study were to assess the frequency and tissue distribution patterns of the inhibin subunits in normal human endometrium. Samples from human endometrium from proliferative phase (PP; n=32), early secretory phase (ES; n=10) and late secretory phase (LS; n=12) were obtained. Immunohistochemistry, immunofluorescence and a statistical analysis were performed. All three inhibin subunits were expressed by normal endometrium by immunohistochemistry and immunofluorescence. Inhibin- was primarily detected in glandular epithelial cells, while inhibin- subunits were additionally localised in stromal tissue. Inhibin- staining reaction increased significantly between PP and ES (P<0.05), PP and LS (P<0.01), and ES and LS (P<0.02). Inhibin-A and -B were significant higher in LS than PP (P<0.05) and LS than ES (P<0.05). All three inhibin subunits were expressed by human endometrium varying across the menstrual cycle. This suggests substantial functions in human implantation of inhibin- subunit, while stromal expression of the subunits could be important in the paracrine signalling for adequate endometrial maturation. The distinct expression in human endometrial tissue suggests a synthesis of inhibins into the lumen and a predominant secretion of activins into the stroma.I. Mylonas and U. Jeschke contributed equally to this work     

人工合成寡肽对大鼠黄体细胞孕酮生成的调节及其可能作用机制

本实验比较了合成寡肽抗孕酮生成作用,并进行了相应机制探讨。发现当ＰＨ７．３－７．５时,能较强抑制孕酮分泌的寡肽其结构有共同特点;活性寡肽可对ＰＬＣ信使传递系统产生抑制作用。也可能通过调节黄体细胞内钙离子浓度降低了ｈＣＧ致孕酮的生成作用,甘－丝－赖还升高黄体细胞中ＰＫＣ活性,而降低了ＰＫＡ。可见人工合成寡肽的抗孕酮作用分子机制十分复杂,有待于深入探讨。     

Secretory Pattern of Inhibin During Estrous Cycle and Pregnancy in African (Loxodonta africana) and Asian (Elephas maximus) Elephants

The ovary of female elephants has multiple corpora lutea (CL) during the estrous cycle and gestation. The previous reports clearly demonstrated that inhibin was secreted from lutein cells as well as granulosa cells of antral follicles in cyclic Asian elephants. The aim of this study is to investigate the inhibin secretion during the pregnancy in African and Asian elephants. Two African elephants and two Asian elephants were subjected to this study. Circulating levels of immunoreactive (ir‐) inhibin and progesterone were measured by radioimmunoassay. Four pregnant periods of an African elephant and three pregnant periods of an Asian elephant were analyzed in this study. Circulating levels of ir‐inhibin started to increase at 1 or 2 week before the ovulation and reached the peak level 3 or 4 weeks earlier than progesterone during the estrous cycle in both African and Asian elephants. After last luteal phase, the serum levels of ir‐inhibin remained low throughout pregnancy in both an African and an Asian elephant. The mean levels of ir‐inhibin during the pregnancy were lower than the luteal phase in the estrous cycle despite high progesterone levels were maintained throughout the pregnancy. These results strongly suggest that CL secrete a large amount of progesterone but not inhibin during the pregnancy in elephants. Zoo Biol 31:511‐522, 2012. © 2011 Wiley Periodicals, Inc.     

The Role of Tissue-type Plasminogen Activator in Rat Corpus Luteum

It is here reported for the first time that luteal cells are capable of secreting plasminogen activators(PA),(both tissue-type,tPA,and urokinase-type,uPA),and plasminogen activator inhibitor type-1(PAl-1).Using organ culture model,we have demonstrated that tPA,but not uPA,showed markedchange during luteolytic period in rat corpus luteum.A great amount oftPA was secreted in corpusluteum on D 14 and D 17 while very low level of tPA activity was detected before D 12.Correspondingly,the progesterone production in the corpus luteum increased gradually in a time-dependent manner from D 1 to D 12 but dropped abruptly to a very low level on D 14.Additionof exogenous tPA to the CL culture caused considerable decrease in progesterone secretion whileinclusion of purified monoclone tPA antibodies in the culture augmented progesterone productionof CL.It is therefore suggested that tPA may play an important role in luteolytic process.     

MMP-26 在人正常胎盘滋养层细胞中的表达及激活素 A 对其表达的调节

胚胎植入和胎盘形成涉及细胞外基质的降解和重建,以及细胞的增殖、凋亡、迁移和分化,基质金属蛋白酶 (MMPs) 是参与这些事件的主要蛋白水解酶系统 . MMP-26 是近年来发现的 MMPs 家族的新成员,但其功能所知甚少 . 通过半定量 RT-PCR 、免疫组织化学、荧光免疫细胞化学等手段,发现人胎盘中 MMP-26 主要定位于绒毛滋养层细胞,在绒毛间质细胞中也有少量表达 . 妊娠早期,胎盘中 MMP-26 表达水平较高,至妊娠中期降至最低,但在足月胎盘中其表达又有显著提高,提示 MMP-26 可能参与妊娠早期滋养层细胞的侵润和分娩时的胎盘剥离 . 体外培养的妊娠早期人细胞滋养层细胞能产生一定水平的 MMP-26 ,而其表达受到激活素 A 的剂量依赖性刺激,表明滋养层细胞中存在 MMP-26 表达的自分泌 / 旁分泌调节 .     

用构建的新型BmNPV载体在家蚕高效表达人β-干扰素

家蚕核多角体病毒(Bombyx mori Nuclear Polyhedrosis Virus.BmNPV)和家蚕细胞已成功地用来大量生产具有生物活性的重组蛋白。但是BmNPV的通用载体的类型较少。因此,本实验构建了BmNPV新型载体pBm92,该载体将多角体蛋白基因的起始密码ATG改变为ATT,然后在多角体蛋白基因的 12位外连接有5个外源基因的克隆位点。将HuIFN-β基因克隆在多角体蛋白基因的 12位后,构建了pBmIFN 12;同时构建HuIFN-β克隆在-3位后的转移栽体pBmIFN-3。将两种转移载体DNA分别与BmNPV基因组DNA共转染Bm-N细胞。利用重组病毒不产生多角体蛋白的特征,筛选重组病毒。用HuIFN-β基因探针与重组病毒DNA进行杂交鉴定。重组病毒BmIFN 12感染Bm-N细胞,其上清IFN活性最高时可达2.0×10~6IU/ml,将BmIFN 12注射5龄家蚕虫体,表达水平为50×10~7IU/ml,是HuIFN-β基因克隆在多角体蛋白基因的-3位后获得的重组病毒的表达量的2～4倍。家蚕体生产的rHulFN-β为糖基化蛋白具有天然HuIFN-β的抗原性。