激活素A和抑制素A对人绒毛膜上皮癌细胞系的增殖及分泌孕酮的影响

通过比较两种肽类激素对细胞增殖与激素分泌的作用，了解它们的作用方式的差异。以来源于人胎盘的绒毛膜上皮癌细胞系Ｊａｒ细胞为实验材料，利用ＭＴＴ法测定细胞增殖，利用放射免疫法测定培养液中的孕酮含量，研究了激活素Ａ和抑制素Ａ对Ｊａｒ细胞的增殖和孕酮分泌的影响。     

γ-氨基丁酸对离体大鼠黄体化颗粒细胞孕酮分泌的影响

用离体孵育方法,观察γ-氨基丁酸（GABA）对大鼠黄体化颗粒细胞孕酮分泌的影响,同时观察在PMSG、蝇覃醇、放线菌酮、forskolin作用下,GABA对黄体化颗粒细胞孕酮分泌的影响。结果显示GABA促进大鼠黄体化颗粒细胞孕酮分泌,蝇覃醇有同样作用。GABA减弱PMSG对颗粒细胞孕酮分泌的促进作用。GABA在放线菌酮作用下对颗粒细胞孕酮分泌无显著影响。GABA促进forskolin作用下的颗粒细胞孕酮分泌。提示GABA促进大鼠黄体化颗粒细胞孕酮分泌是通过结合到颗粒细胞上的GABA受体,可能与细胞内腺苷酸环化酶系统有关,而与蛋白质合成无关。     

表皮生长因子、肾上腺素及前列腺素E_2对大鼠离体黄体细胞孕酮分泌的影响

用放射免疫分析法测定了离体培养的雌性大鼠黄体细胞孕酮分泌及表皮生长因子(EGF)、肾上腺素(E)、前列腺素E_2(PGE_2)对孕酮分泌的影响。结果表明:表皮生长因子(2～200ng)抑制黄体细胞孕酮的基础分泌及hCG诱导孕酮分泌,并呈量效关系;肾上腺素在50μg/ml时刺激孕酮分泌;表皮生长因子能阻断肾上腺素的促进作用;前列腺素E_2则抑制黄体细胞孕酮分泌,既抑制基础分泌又抑制hCG诱导的孕酮分泌。     

某些氨基酸及小分子肽对大鼠卵泡颗粒细胞雌激素与孕激素分泌的影响

用离体大鼠卵泡颗粒细胞培养和放射免疫分析法,观察酪氨酸(Tyr)、丝氨酸(Ser)、苏氨酸(Thr)和八种新合成的小分子肽(Gly-Tyr-NH_2,Lys-Tyr-NH_2,Gly-Gly,Gly-Tyr,Gly-Tyr-Lys,Gly-Ser-Lys,Thr-Ser-Lys,Thr-Ser-Tyr)对雌激素和孕激素分泌的影响。结果表明,Tyr和Thr对颗粒细胞基础雌二醇分泌有很强的刺激作用;Tyr、Ser、Thr对颗粒细胞hCG引起的雌二醇分泌没有影响。上述八种合成小分子肽对hCG引起的雌二醇分泌均有增强作用,其中二肽Gly-Tyr和三肽Gly-Ser-Lys对基础雌二醇分泌有促进作用。小分子肽对颗粒细胞孕酮分泌的影响比较复杂,Gly-Tyr-NH_2和Thr-Ser-Tyr抑制基础孕酮分泌;Gly-Tyr-NH_2、Gly-Ser-Lys和Gly-Tyr增强hCG致孕酮分泌,其余几种小分子肽对孕酮分泌没有影响。     

白细胞介素-1、2、6对大鼠离体黄体细胞孕酮生成的影响

近年来研究证实白细胞介素一１（ＩＬ－１）、白细胞介素－２（ＩＬ－２）和白细胞介素－６（ＩＬ－６）可通过自分泌或旁分泌的方式影响生殖机能的各个环节,包括卵巢功能、子宫内膜周期变化、胚胎发育及胎盘功能等。本研究着重探讨了ＩＬ－１、ＩＬ－２和ＩＬ－６对黄体细胞孕酮生成的影响。 取２１－２５天龄未成年雌性大鼠,经ＰＭＳＧ－ｈＣＧ处理后,取黄体组织用胶原酶和 ＤＮ Ａ酶消化、制成黄体细胞悬液、加合２％小牛血清的Ｍ１９９培养液置３７℃ ５％的ＣＯ２培养箱预培养２４ｈ,冲洗后换新鲜无血清Ｍ１９９培养液及不同浓度的…     

半乳糖对小鼠胚胎发育的影响

目的研究半乳糖对妊娠小鼠胎盘组织脂质过氧化和抗氧化酶的影响，探讨半乳糖对小鼠胚胎发育的影响及其机理。方法将75只妊娠小鼠随机分为4组，即半乳糖低、中、高浓度组（20％、35％和50％）和正常对照组，分别从妊娠第1d开始喂不同浓度半乳糖（20％、35％、50％）及正常食物，观察妊娠结局，记录总胎鼠数、活胎数、吸收胎数和死胎数。测定35％半乳糖组孕鼠胎盘组织中超氧化物歧化酶（SOD）的活性及脂质过氧化产物丙二醛（MDA）含量。采用放免法（RIA）测定35％半乳糖组孕鼠及对照组孕d5、d9、d13和d15血清孕酮（P）和雌二醇（E2）水平。结果（1）35％半乳糖可使妊娠小鼠血清孕酮和雌二醇水平较对照组显著下降（P〈0．05）（2）35％半乳糖使妊娠小鼠胎盘组织中SOD活性较对照组显著降低，MDA含量较对照组显著升高（P〈0．01）（3）随着半乳糖浓度增高，妊娠小鼠活胎率明显下降，吸收胎率和死胎率明显升高（P〈0.05）。结论半乳糖致妊娠小鼠胎盘组织中抗氧化能力下降，氧化水平增强，破坏胎盘的抗氧化系统的平衡状态，造成胎盘组织氧化损伤，使发育中的胚胎失去血液供应和营养，最终导致胚胎发育受阻，孕体被排出或吸收。     

人早孕绒毛滋养层细胞的分离纯化及马鞭草抗早孕机理的初步研究

目的：探讨马鞭草抗早孕的细胞学作用机理。方法;正常妊娠６－８周早期胎盘Ｐｅｒｃｏｌｌ梯度离心分离出滋养层细胞进行体外培养,观察马鞭草对培养的滋养层细胞形态及绒毛膜促性腺激素分泌功能的影响。结果：２５,５０ｍｇ／ｍｌ马鞭草乙醇提取液对滋养层细胞生长及绒毛膜促性腺激素分泌有明显的抑制作用。结论;一定浓度的马鞭草可直接杀伤滋养层细胞,抑制绒毛膜促性腺激素的分泌而中止早孕。     

磷酸化寡肽(P-GT)对离体培养大鼠黄体细胞孕酮分泌的影响

本文观察了人工合成的磷酸化寡肽甘氨酸酪氨酸胺（合成代号P-GT）对黄体细胞孕酮分泌的影响，并对其机制进行了初步探讨，结果显示：该寡肽对黄体细胞孕酮基础分泌和人绒毛膜促性腺激素（hCG）诱导下的黄体细胞孕酮分泌都具有显著促进作用，且与腺苷酸环化酶激动剂Forskolin有协同作用，提示其可能是通过腺着酸环化酶——蛋白激酶A（AC－PKA）信息系统发挥作用，Ca2 对该寡肽促孕酮作用可能没有影响。     

抑制和兴奋性氨基酸对兔卵巢颗粒细胞雌孕激素生成的影响

本实验采用离体孵育的兔卵巢颗粒细胞，观察四种氨基酸神经递质对颗粒细胞雌二醇和孕酮生成的影响。结果表明，γ-氨基丁酸和谷氨酸能明显刺激颗粒细胞在基础状况下雌二醉的分泌而抑制孕酮的分泌；甘氨酸和天门冬氨酸无此作用。γ-氨基丁酸、甘氨酸和谷氨酸明显促进hCG诱导的颗粒细胞雌二醇的分泌和抑制孕酮的分泌而；天门冬氨酸无此作用。发挥作用的机制可能与腺苷酸环化酶-cAMP系统有关。     

乙醇刺激HepG2细胞分泌VASN蛋白的生物学功能研究

目的：探究乙醇对HepG2细胞VASN蛋白表达与分泌水平的影响,及其对HUVEC细胞迁移的影响。方法乙醇刺激HepG2细胞,定量PCR （ qPCR）检测HepG2细胞VASN mRNA水平的变化, Western印迹检测HepG2细胞与细胞上清中VASN蛋白水平的变化。划痕愈合实验检测细胞共培养条件下,乙醇刺激的HepG2细胞培养上清对HUVEC细胞迁移的影响。结果 qPCR结果显示,乙醇可上调HepG2细胞VASN mRNA水平。 Western印迹结果显示,细胞VASN蛋白表达与分泌水平升高。划痕愈合实验结果表明乙醇刺激HepG2细胞VASN蛋白的分泌增加,可以促进HUVEC细胞的迁移。结论乙醇刺激导致HepG2细胞VASN蛋白表达水平和分泌水平升高,并可促进与其共培养的HUVEC细胞迁移。     

The somatostatin analogue, octreotide, modifies both steroidogenesis and IGFBP-1 secretion in human luteinizing granulosa cells

The effect of the somatostatin analogue octreotide on the secretion of progesterone and insulin-like growth factor binding protein-1 (IGFBP-1) from human granulosa-luteal cells was investigated. Octreotide (10(- 11), 10(-10) and 10(-9) M) alone induced a significant decrease in progesterone secretion (maximum suppression 69 +/- 5% of control: P < 0.0001). In contrast, treatment with octreotide in combination with human chorionic gonadotrophin (HCG), at 1 IU/ ml potentiated the stimulatory effect of HCG on progesterone secretion (HCG alone 201 +/- 3% of control: HCG + octreotide 10(-9) M 318 +/- 16%: P < 0.001). Treatment with octreotide increased the secretion of IGFBP-1 (maximum stimulation 254 +/- 25% of control: P < 0.01). No effect of HCG was seen on secretion of IGFBP-1. These findings raise the possibility that somatostatin may have a modulatory role in regulating steroidogenesis by the human corpus luteum. Further studies are required to establish the physiological significance of any such function.      

Circhoral fluctuations of serum total renin, inhibin and related hormones around the mid-cycle in normal human females.

Total renin and inhibin are secreted by the ovary. Although luteinizing hormone (LH) and/or follicle stimulating hormone (FSH) may stimulate their secretion, the close relationship between fluctuations of gonadotrophins, oestradiol, progesterone, renin and inhibin during the cycle is still conjectural. To investigate the temporal relationship between the short-term fluctuations in the circulating concentrations of LH and FSH and the ovarian hormones (oestradiol, progesterone, renin and inhibin), blood samples were collected at 15-min intervals for 6 h from 15 normal women in the late follicular (n = 4), early luteal (n = 5) or luteal (n = 6) phases of the menstrual cycle. LH levels showed the well-known pulsatile secretion with decreasing frequency and increasing relative amplitude from the late follicular to the luteal phase. Progesterone and oestradiol serum levels were pulsatile, 25% and 35-50% of which were linked to LH pulses, with time lags of 30 and 12-15 min respectively. Renin levels presented significant pulses, 26% of which were related to LH pulses with a time lag of less than 10 min; no coincidence was found between renin and oestradiol pulses. Inhibin levels presented only scattered pulses of small amplitude, which were unrelated to LH or FSH. These results show that, besides the LH-related pulses, pulsatile secretion of some ovarian hormones (oestradiol, progesterone and renin) may also occur independently of LH pulses and may be unrelated to one another. Moreover, contrary to the other ovarian hormones, inhibin seems to follow a tonic, not a pulsatile type of secretion around the mid-cycle.     

Regulatory role of angiotensin II on progesterone production by cultured human granulosa cells. Expression of angiotensin II type-2 receptor

The role of angiotensin II (AngII) in ovarian steroidogenesis is not clearly understood. In order to study its action on progesterone synthesis and to determine which receptor subtype is involve, granulosa cells obtained from women undergoing in-vitro fertilization were cultured for 2 or 4 days and then incubated in the presence of AngII (10(-7) M) with or without human chorionic gonadotrophin (HCG, 10 IU/ml) for 3 or 18 h. In cells cultured for 2 days, incubation with AngII decreased progesterone secretion by 36%, and inhibited activity of 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) by 87% (P < 0.05), although its expression was not significantly reduced. However, in cells cultured for 4 days, progesterone production was enhanced by incubation with AngII (38%), and no change was observed in 3 beta-HSD expression. Both inhibitory and stimulatory effects were dose- dependent. Progesterone secretion was increased (93%) by incubation with HCG of cells cultured for 4, but not for 2 days, and no potentiation was observed with AngII. Treatment with PD123177 completely blocked the action of AngII and decreased the HCG-stimulated secretion of progesterone by 27%. Angiotensin type-2 (AT2) receptor mRNA was expressed in cells cultured for 4 days. In conclusion, AngII showed a regulatory role in in-vitro progesterone production by human granulosa luteinized cells, modulating the activity of 3 beta-HSD. It is likely that these actions may be mediated via membrane receptors, possibly of the AT2 receptor family.      

Support for the hypothesis that successful immunotherapy of various cancers can be achieved by inhibiting a progesterone associated immunomodulatory protein

A hypothesis was proposed that cancer cells may utilize a pre-existing mechanism that pregnant mammals use to avoid natural killer cell immune surveillance of the fetus. The hypothesis suggested that those cancer cells that are able to proliferate may have found a way to cause the expression of the immunomodulatory protein known as the progesterone induced blocking factor (PIBF). The cancer cells could find an alternate pathway to make this protein that does not require progesterone secretion, or the cancer cells may actually utilize progesterone and thus make PIBF in a similar fashion to normal pregnancy. If the former mechanism was operational, then one could develop monoclonal antibody type therapy directed to this unique protein not needed for normal body functions. However, if the latter pathway involving progesterone secretion is operational, then there would be drugs already on the market, e.g., the progesterone receptor antagonist mifepristone that could be used to treat these cancers. In vitro data has shown that 100% of human leukemia cell lines express mRNA for the PIBF protein. Some leukemia cell lines have been found that actually express the PIBF protein. In fact adding progesterone to the culture media upregulated PIBF protein expression and mifepristone inhibited it. Controlled studies in various murine spontaneous cancers not known to be associated with progesterone receptors showed increased length and quality of life following mifepristone therapy. Anecdotal improvement in advanced widely metastatic human cancers has also been found. Thus there is now experimental data to support this hypothesis and a new door to a completely different type of cancer therapy has been opened.     

Effects of androstenedione, insulin and luteinizing hormone on steroidogenesis in human granulosa luteal cells

BACKGROUND: The present study was conducted to investigate the effect of androstenedione, insulin and LH on human granulosa cell oestrogen and progesterone production. We postulated that elevated concentrations of androstenedione, insulin and LH may be important modulators of granulosa cell steroidogenesis. METHODS: Granulosa cells obtained in connection with IVF procedures were cultured for a total of 4 days in serum-free medium containing androstenedione (10(-6) mol/l). We tested the effect of androstenedione (10(-5) mol/l) on insulin (0-800 microIU/ml), LH (1-10 ng/ml) as well as on insulin + LH-stimulated oestrogen and progesterone production. RESULTS: Insulin increased the basal secretion of steroid hormones, and furthermore augmented LH-stimulated oestrogen and progesterone accumulation in granulosa cell cultures. Androstenedione (10(-5) mol/l) stimulated basal oestrogen production, but significantly reduced (32-58%) insulin + LH-stimulated oestrogen and progesterone secretion (P < 0.05). CONCLUSION: These results suggest that high androstenedione concentrations may act directly to impair insulin augmentation of LH-stimulated oestradiol and progesterone production in cultured human granulosa luteal cells. This is compatible with the hypothesis that high androgen levels may inhibit oestrogen production in polycystic ovary follicles, and as such may contribute to anovulation and infertility in women with polycystic ovary syndrome.     

In-vivo administration of progesterone inhibits the secretion of endometrial leukaemia inhibitory factor in vitro

Human interleukin for DA cells, also called leukaemia inhibitory factor (LIF), is of cardinal importance for successful murine embryo implantation. Recent studies suggest it may also play an important role in human embryo implantation. Our objective was to study the hormonal regulation of the production/secretion of LIF by the human endometrium. Endometrial LIF secretion in specimens obtained from women without ovarian function (n+/-14) at day 10 (4 mg of oestradiol regimen) or day 20 (oestradiol plus 300 mg of progesterone) of a simulated menstrual cycle was examined. LIF was detected in all cultured explants obtained both in the proliferative and secretory phase of the stimulated cycles. The levels of cytokine production by day 10 endometrial culture explants were 5-fold higher than by day 20 endometrial samples (mean+/- SEM, 24.3+/-8.6 versus 4.5+/-2.1 pg/mg, P < 0.01). This suggests that progesterone significantly down-regulates the endometrial LIF secretion. The effect of progesterone on LIF secretion by the endometrium in vitro was also examined. Explants of endometrium obtained from the same patients on day 10 of cycle were treated with 0.5 ng/ml of progesterone in vitro. This progesterone treatment significantly reduced LIF secretion by endometrial explants in vitro (mean+/-SEM, 20.3+/-4.8 versus 10.7+/-2.3 pg/mg, P < 0.05). These results suggest that LIF endometrial production is regulated by progesterone both in-vivo and in-vitro. The possible mechanisms of LIF regulation are discussed briefly.      

Interleukin-17 Increased Progesterone Secretion by JEG-3 Human Choriocarcinoma Cells

Problem  JEG-3 choriocarcinoma cell line has previously been reported to express a receptor for interleukin (IL)-17. The involvement of IL-17 in the production of progesterone and human chorionic gonadotropin by placental trophoblast has not been investigated.
Method of study  The present study investigated the in vitro effect of IL-17 on progesterone and human chorionic gonadotropin (hCG) secretion by JEG-3 cells. Both hormones were quantified using enzyme-linked immunosorbent assays.
Results  The results showed that IL-17 significantly increased progesterone secretion at 6 ( P  < 0.001) and 24 ( P  < 0.01) hr, while this cytokine had no effect on hCG secretion.
Conclusion  Interleukin-17 may regulate the function of JEG-3 cells through increased progesterone secretion.     

GABA抑制hCG诱发大鼠卵巢黄体细胞孕酮生成

观察了中枢抑制性神经递质γ 氨基丁酸 (GABA)对人绒毛膜促性腺激素 (hCG)诱发大鼠卵巢黄体细胞孕酮的生成 ,及对羟自由基 (·OH)产生的影响。结果显示 :GABA抑制基础及hCG诱发黄体细胞孕酮的生成 ,同时发现黄体细胞羟自由基的生成增多。表明 :GABA抑制孕酮的生成可能与自由基有关 ,并可能通过cAMP PKA系统发挥作用 ,与GABA在中枢神经系统的作用机制不同     

Both iso- and hyperosmotic ethanol stimulate release of hypothalamic thyrotropin-releasing hormone despite opposite effect on neuron volume

Previous studies have indicated that isosmolar, but not hyperosmolar, ethanol induces in vitro gonadotropin-releasing hormone secretion from the basal hypothalamus, presumably by causing cell swelling. Moreover, ethanol reduces secretion of another hypothalamic neuropeptide vasopressin. We have studied the acute effect of ethanol on specific hypophysiotropic basal and K⁺-stimulated thyrotropin-releasing hormone secretion in vitro especially in relation to cell swelling. Isosmotic 40–160 mM ethanol increased thyrotropin-releasing hormone release from the hypothalamic paraventricular nucleus and median eminence in a dose-dependent manner. Both a 30% decrease of osmolarity and isosmotic 80 mM ethanol induced 12% swelling of hypothalamic neurons. Hyperosmotic 80 mM or 160 mM ethanol induced release of thyrotropin-releasing hormone from both hypothalamic structures but did not cause cell swelling (80 mM) or even induced cell shrinkage (160 mM). Depletion of medium Ca²⁺ did not affect thyrotropin-releasing hormone secretion caused by either isosmotic or hyperosmotic ethanol.

Our data indicate that both iso- and hyperosmotic ethanol stimulated release of hypophysiotropic thyrotropin-releasing hormone despite opposite effects on neuron volume. The mechanism of ethanol action appears complex and variable depending on the type of cell and neuropeptide affected.     

The inhibitory action of ethanol on the gastric mucosa and its interaction with the vagus in rats

The influence of systemic ethanol and/or the vagus on ionic secretion and on glandular mucosal blood flow (GMBF) was studied in anex-vivo gastric chamber preparation in rats. Sub-diaphragmatic vagotomy decreased H⁺ secretion and Na⁺ outflux from the gastric mucosa. Subcutaneous injection of 50% ethanol significantly potentiated these responses, but not the concentrations of 25% and 100%. The three doses of ethanol did not affect the secretion of both H⁺ and Na⁺ in vagus-intact animals. Ethanol, however dose-dependently reduced the GMBF in both vagus-intact and sub-diaphragmatic vagotomised rats, and the effect was greater in the latter-operated animals. It is concluded that vagus nerves greatly influence the secretion of both H⁺ and Na⁺ the gastric mucosa but the effect is unrelated to GMBF. Systemic ethanol reduced the secretion of these ions only in vagotomised animals, indicating the vagus could play a role in modulating the action of ethanol in the stomach.