Expression and subcellular localization of urocortin in syncytiotrophoblast of human term placenta

AIM: To obverse the expression and localization of urocortin on ultrathin cryosections of syncytiotrophoblast of human term placenta with immunocytochemistry technique under transmission electron microscope. METHODS: The human term placenta tissue from Cesarean delivery and normal labor were fixed in 4% paraformaldehyde, and then divided into two parts. One part was for regular immunocytochemistry under microscope, and the other part was used to prepare ultrathin cryosections for immunocytochemistry under transmission electron microscope. RESULTS: 1.Uroncortin mainly distributed in cytoplasm of syncytiotrophoblast of human term placenta under microscope. Urocortin also appeared in cytoplasm in some stromal cells. 2. Under transmission electron microscope, the anti-urocortin gold particles were observed in cytoplasm of syncytioptrophoblast ultrathin cryosections and sited on rough-surfaced endoplasmic reticulum. The anti-urocortin gold particles also appeared on nucleus and nuclear membrane of syncytiotrophoblast. CONCLUSION: Syncytiotrophoblast of human term placenta synthesized and secreted urocortin. The internalization of urocortin within syncytiotrophoblast nuclear indicates that urocortin may act as intracrine.     

牛羊胎盘免疫调节因子的提取及对PANC-Ⅰ增殖和迁移影响的比较

胎盘免疫调节因子(placenta immunoregulating factor,PF)是从胎盘提取的小分子多肽,对病毒性疾病、免疫缺陷疾病及恶性肿瘤等具有潜在的临床应用价值.为检测牛(Bos taurus)和绵羊(Ovis aries)胎盘免疫调节因对人(Homo sapiens)胰腺癌细胞(pancreatic cancer cells-Ⅰ,PANC-Ⅰ)增殖与迁移的影响,本研究对从屠宰场获得的牛和绵羊胎盘进行分离,利用双酶水解法制备PF,将质量浓度为100～2 000 ng/mL的牛、羊PF作用于体外培养的PANC-Ⅰ细胞,以3-(4,5-二甲基噻唑-2-基)-5-(3-羧甲酯基)-2-(4-磺苯基)-2H-四唑(3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium,MTS)法观察细胞增殖抑制率,应用流式细胞技术检测其对PANC-Ⅰ细胞周期的影响.并利用Transwell迁移实验检测添加PF对PANC-Ⅰ细胞迁移能力的影响,同时通过实时荧光定量PCR法检测肿瘤抑制基因(tumor suppressor gene,PS3)和细胞周期依赖性蛋白激酶抑制因子1A基因(cyclin-dependent kinaseinhibitor 1A,P21)的表达情况.结果显示,17.11 g绵羊胎盘制得0.588 mg PF,6.61 g牛胎盘制得0.312 mgPF,胎盘质量与所得PF的质量比约为20 000∶1.在体外培养的PANC-Ⅰ细胞培养液中添加PF,从形态上观察,牛羊PF对PANC-Ⅰ细胞没有明显的影响.绵羊PF处理后PANC-Ⅰ细胞增殖抑制率为9.75％～22.89％,牛PF处理后PANC-Ⅰ细胞增殖抑制率为-3.81％～23.56％,增殖抑制率最显著的绵羊PF浓度为500 ng/mL,牛PF浓度为1 000 ng/mL.绵羊PF在浓度为500 ng/mL时,PANC-Ⅰ细胞的PI和SPF最低,而牛PF各浓度下的胰腺癌细胞增殖指数(proliferation index,PI)和增殖活性(S-phase fraction,SPF)没有明显的变化.同时,牛羊PF对PANC-Ⅰ细胞的迁移能力无显著影响.牛羊PF使PANC-Ⅰ细胞P53表达上调而P21表达下调.上述结果表明,在一定浓度下,PF对体外培养的PANC-Ⅰ细胞增殖有负调控作用,且牛羊胎盘免疫调节因子对胰腺癌细胞的影响不同,可能因为其存在的活性成分有区别.本研究为胰腺癌的治疗提供了新的思路.     

Influence of transection of the cervical sympathetic trunk on content of NO and expression of NOS in placenta of rats with pregnancy-induced hypertension syndrome

AIM:To observe the influence of transection of the cervical sympathetic track (TCST) on the content of NO and the expression of eNOS mRNA and iNOS mRNA in placenta of the rats with pregnancy-induced hypertension syndrome (PIH).METHODS: Pregnant Wistar rats were randomly divided into 5 groups: control group (C): saline was injected subcutaneously from 14th day to 20th day of gestation;PIH group 1 (H1) and group 2 (H2): L-NAME was respectively injected with 125 mg/kg and 62.5 mg/kg,respectively,then the other procedures were the same as group C;Operation group (O): TCST was operated on 14th day of the gestation,then the other procedures were the same as group H1;sham operation group (S): the cervical sympathetic trunk was only separated and exposed on 14th day of the gestation,then the other procedures were the same as group H1.RESULTS: (1) Except the base value of the BP and protein in urine of the pregnant rats,all the parameters observed in group H1 and H2 were higher than those in group C significantly (P<0.01),and in group H2 were lower than those in group B1 markedly (P<0.01).(2) In comparison with those in group C,the size and body weigh of fetus in group H1,H2 decreased markedly (P<0.01).The above indexes in group H1 were lower than those in group H2 markedly (P<0.01,P<0.05).The changes of the rate of embryo absorption and fetal death,and deformity rate of the fetal rats were contrary to the above indexes.(3) The content of NO and the expression of eNOS mRNA and iNOS mRNA in placenta in group H1 and H2 were lower than those in group C markedly (P<0.01).Those in group H1 were lower than those in group H2 obviously (P<0.01,P<0.05).Those in group O were higher than those in group H1 markedly (P<0.01).CONCLUSION: TCST protects pregnant rats against PIH,and it was related to the mRNA expression of eNOS and iNOS and the content of NO in placenta tissue.     

Changes in Placental Hormone Secretion After Dexamethasone Treatment in Cattle

Contents Catheters were surgically placed in the uterine artery and vein of four control and four dexamethasone (DXMS)-treated cows on days 248 or 249 of gestation; at the same time the corpus luteum was removed and all animals were treated with 10 mg chlormadinone acetate i.m. daily to maintain pregnancy. After one day of blood sampling which began on day 250, animals received either 20 mg DXMS crystalline suspension i.m. or only the vehicle solution. Blood sampling was then continued for five additional days. Plasma was assayed for unconjugated estrone (E1), progesterone (P4), and pregnenolone (P5). Concentrations of El were higher in the venous samples (p < 0.02) and increased over time (p < 0.02). There was a site x time interaction (p < 0.05) and a significant effect of DXMS-treatment on the linear component (p < 0.02) of the slope values for the El concentrations for both sampling sites. Concurrently, there was an effect of time on P4 concentrations (p lt; 0.01), the venous P4 concentrations were higher (p < 0.01), and there was also a site x time interaction (p < 0.01) but not treatment effect. There was no significant effect on concentrations of P5 between treatment goups or sampling sites. It is concluded that DXMS activates the synthetic system(s) involved in production of estrone in the cow during late pregnancy. This change in estrone synthesis occurred at an increasing rate during the 5 day sampling period. The data are consistent with the hypothesis that the placenta must be capable of utilizing precursors in the cholesterol or pregnenolone group for estrone synthesis and that the major pathway could be the delta-5 pathway. Inhalt: Veränderungen in der placentären Hormonsekretion beim Rind nach Behandlung mit Dexamethason. Bei insgesamt 8 Kühen wurden am 248./249. Tag der Trächtigkeit Katheter in die Vena und Arteria uterina eingelegt, gleichzeitig wurde das Corpus luteum entfernt und zur Aufrechterhaltung der Gravidität wurden 10 mg Chlormadinonacetat täglich während der gesamten Versuchsdauer i.m. verabreicht. Nach einer ersten Blutentnahmeperiode am Tag 250 erhielten jeweils 4 Tiere entweder 20 mg Dexamethason (kristalline Suspension) bzw. das Vehikel (Kontrollguppe) i.m. verabreicht. Die Blutentnahmen erfolgten über weitere 5 Tage, erfaßt wurde der Verlauf von freiem Estron, Progesteron und Pregnenolon. Die in der Vena uterina gemessenen Estronkonzentrationen lagen jeweils höher (p < 0,02) als in der entsprechenden Arterie und stiegen bis zum Ende des Experiments an (p < 0,02). Weiterhin zeigte sich eine Interaktion zwischen dem Ort der Blutentnahme und dem Zeitpunkt (p < 0,05), der Anstieg der Estronkonzen-trationen nach der Behandlung mit Dexamethason war – bezogen auf die lineare Komponente – sigifikant erhöht (p < 0,02). Für P4 ergab sich ein deutlicher Abfall (p < 0,01), wobei in der Vena uterina stets höhere Werte gemessen wurden als in der Arteria uterina (p < 0,01). Auch hier war eine Interaktion zwischen dem Zeitpunkt und dem Ort der Blutentnahme gegeben (p < 0,01), allerdings konnte ein Behandlungs-effekt nicht dargestellt werden. Für Pregnenolon ergaben sich keine Unterschiede zwischen Arteria und Vena uterina sowie den Behandlungsgruppen. Die Ergebnisse zeigen, daß eine einmalige Behandlung mit Dexamethason zu einer zunehmenden Steigerung der Estronproduktion führt, im Hinblick auf die unveränderte Progesteronsekretion wird geschlossen, daß dabei eher der Delta-5-Syntheseweg zugrundeliegen könnte und daß die entsprechenden Precursoren auf der Ebene des Cholesterins oder Prepenolons zu suchen sind.     

Endothelin-1 concentrations in clone calves, their surrogate dams, and fetal fluids at birth: association with oxygen treatment

Increased endothelin-1 (ET-1) plasma concentration in human infants is associated with persistent pulmonary hypertension of the newborn, a problem also identified in calves derived from somatic cell clone technology. Increased ET-1 also is present in the amnionic fluid and plasma of the infant and mother in preeclampsia, a condition associated with abnormal placentation. Abnormalities in placentation are identified in clone calves. We measured ET-1 in fetal fluid, calf plasma, and surrogate dam plasma in 40 clone calves at the time of term delivery. Calves were subsequently identified as being either oxygen treated (O2) or non-oxygen treated based on their postpartum clinical course. Fetal fluid ET-1 concentration greater than 1.4 ng/mL carried a 3-fold increase in odds of the calf being treated with oxygen. Maternal plasma ET-1 concentration was greater in the O2 group (13 pg/ mL: [8-23 pg/mL] versus 25 pg/mL [12-40 pg/mL]; median, 25-75 percentile). Plasma ET-1 concentration in calves was not significantly different between groups. Fetal fluid ET-1 may serve as a marker for neonatal disorders of oxygenation in clone calves and the source of ET-1 may be the placenta.     

Morphological Characteristics of the Placenta and Umbilical Cord of Arabian Mares Foaling in the United Arab Emirates

A total of 127 normal placentas from Arabian mares resident in the United Arab Emirates were examined. The mean linear dimensions of the placenta were, on average, 84% of those previously recorded for the placentas of the Thoroughbred. Significant differences in the size of the allantochorion between primigravid and multiparous mares were seen only in the linear dimensions of the body portion. The pregnant horn was more commonly on the right than left side of the uterus (P = .01; 74/127; 58%). Cord attachment was primarily at the base of the two placental horns (112/127; 88%), with the remainder showing anomalies from this position. The mean (±SEM) total length of the umbilical cords was 62.2 ± 1.2 cm, being composed of the allantoic portion (29.7 ± 0.9 cm) and amniotic portion (32.5 ± 0.6 cm), which averaged 53 ± 0.01% of the total length. The amniotic portion was usually, but not always, longer than the allantoic portion (79/127; 62%). Longer cords were associated with a greater proportion of allantoic length. An enlarged yolk sac remnant (YSR) was present in 16/127 (13%) placentas. Cords with YSRs displayed a significantly longer allantoic portion than those without (P = .02). The total cord length was not correlated with the weight or area of the allantochorion or amnion, any linear measurement of the allantochorion, gestation length, the month of foaling, parity of the dam, or birthweight or sex of the foal. The purpose for which the mare was bred (i.e., racing or showing) influenced the cord length, those of show mares being significantly longer.