17 beta-hydroxysteroid dehydrogenase activity in leiomyoma and myometrium and its relationship to concentrations of oestrone, oestradiol and progesterone throughout the menstrual cycle

The activity of the enzyme 17 beta-hydroxysteroid dehydrogenase (17 beta-OHSD) and concentrations of oestrone (E1), oestradiol (E2) and progesterone have been measured in leiomyoma and myometrium obtained at different stages of the menstrual cycle. Apart from conversion of E2 to E1 in the proliferative phase, no significant difference in enzyme activity was noted between normal and tumour tissue. However, interconversion in both tissues was shown to be higher in the secretory than the proliferative phase of the menstrual cycle. E1 concentrations were significantly higher (P less than 0.01) in leiomyoma than in myometrium, obtained during the proliferative phase. Concentrations of both oestrogens, in some tumour and normal tissues, were higher in the proliferative than the secretory phase. Secretory phase tissues contained higher concentrations of progesterone than those obtained in the proliferative phase of the menstrual cycle. Considerable differences in both enzyme activity and steroid concentrations were noted in different areas of the same tumour.     

输卵管妊娠时输卵管壁肥大细胞的研究

目的　探讨肥大细胞在输卵管妊娠中的数量变化及其与血清性激素的关系。方法 :取输卵管妊娠时的输卵管及月经周期的增生期、分泌期和正常宫内早孕时的输卵管 ,常规石蜡切片 ,用甲苯胺蓝染色法显示肥大细胞 ;用酶免疫分析法检测输卵管妊娠患者、正常育龄未孕妇女 (增生期和分泌期 )及正常宫内早孕妇女血清雌二醇和孕酮水平。结果 :输卵管妊娠患者血清雌二醇和孕酮水平均高于正常育龄未孕妇女 (增生期和分泌期 ) ,低于正常宫内早孕妇女 ,四组间两两比较差异均有显著性 (P <0 0 5 ) ;肥大细胞主要分布于输卵管肌层 ,其数量变化为 :输卵管妊娠组较增生期和分泌期这两组均少 ,差异均有显著性 (P <0 0 5 ) ,而增生期和分泌期这两组肥大细胞数量变化不明显 ,差异无显著性 (P >0 0 5 ) ;两例正常宫内早孕时的输卵管壁肥大细胞数量明显比输卵管妊娠组多 ,与增生期和分泌期这两组比较 ,肥大细胞数量变化不明显。结论 :1 人输卵管壁内肥大细胞的数量不受血清性激素水平的影响。 2 输卵管妊娠时肥大细胞数量减少     

Protein synthesis and secretion by the human endometrium during the menstrual cycle and the effect of progesterone in vitro

To identify markers of endometrial differentiation specimens of endometrium from the menstrual cycle were incubated in vitro with [35S]methionine, in the absence or presence of progesterone, and protein synthesis and secretion were studied by fluorographic analysis of one dimensional SDS/gradient polyacrylamide gels. Changes were demonstrated in the rate of synthesis and secretion of a number of endometrial proteins (EP) during the cycle and in response to progesterone. Endometrial proteins were classified into three groups: Group I-synthesized and secreted throughout the menstrual cycle and unaffected by progesterone exposure; Group II-synthesis and secretion associated with histological type of endometrium and unaffected by progesterone exposure, e.g. EP 13 (Mr 33,000) with proliferative, EP 15 (Mr 28,000) with secretory and EP 14 (Mr 32,000) with late secretory endometrium; Group III-synthesis and secretion regulated by progesterone exposure irrespective of source of endometrium, e.g. EP 9 (Mr 54,000) and 11 (Mr 45,000). The Group II proteins EP 14 and 15 were also the major secretory protein products of endometrium from first and second trimester pregnancy respectively, the native forms referred to as pregnancy-associated endometrial alpha 1- and alpha 2-globulins (alpha 1- and alpha 2-PEG). We conclude that EP 15 (alpha 2-PEG) represents a human analogue of uteroglobin.     

Tissue culture and estrogen, to clarify the roles of estrone sulfate

Estrone sulfate (E1-S) has been shown to be quantitatively the most important estrogen in peripheral blood. But, the physiological and/or pathological role of E1-S is not yet clarified. At present, we tried to clarify it using tissue cultures. In tissue cultures of human endometrium, secretory endometrium showed higher activity of estrone sulfatase (E1----E1-S) than proliferative endometrium. Progesterone added in the medium induced an increase of estrone sulfotransferase in the proliferative endometrium. The results suggest a reducing effect of estrogen by progesterone in secretory endometrium in physiological conditions. Estrogen dependent malignant tumors (breast cancer, endometrial cancer) have high estrone sulfatase. It converts E1-S to E1 (----E2) which are abundant in these tumors. Ishikawa cell line increased estrone sulfotransferase activity with progesterone, somewhat like the physiological conditions. From out study in vivo, there is a possibility of some ameliorative effects of E1-S on the central nervous system of patients with senile dementia (Alzheimer's type). Effects of E1-S on central nerves were investigated using tissue cultures.     

Immunocytochemical analysis of oestrogen receptors and progesterone receptors in the human uterus throughout the menstrual cycle and after the menopause.

To obtain more insight into the relationship between cyclic and regional changes in steroid receptor expression and function-related changes in the various types of cell of the normal human uterus, we performed an immunocytochemical study on paraffin-embedded sections. The distribution and intensity of immunostaining for the oestrogen receptor and the progesterone receptor in the various types of cell were semiquantitatively scored. The data were statistically compared for the different phases of the menstrual cycle and after the menopause, and for the different regions of the corpus and (endo)cervix uteri. During the menstrual cycle, significant changes in oestrogen receptor score were observed in glandular and stromal cells of endometrium basalis and functionalis and in smooth muscle cells of the myometrium. In all types of cell, oestrogen receptor expression reached a maximum in the late proliferative phase. During the early secretory phase, oestrogen receptor staining declined sharply in stromal and smooth muscle cells, whereas, in glandular epithelium, oestrogen receptor expression decreased more gradually. During mid- and late-secretory phases, an increase in oestrogen receptor staining was also observed in predecidualizing stromal cells and smooth muscle cells. Progesterone receptor numbers changed significantly in glandular epithelium but not in stromal and smooth muscle cells. Glandular progesterone receptor expression reached a maximum in the early secretory phase and was then drastically reduced. During mid- and late-secretory phases stromal cells were moderately stained for progesterone receptor in contrast to epithelial gland cells which showed no or very weak staining. No regional variations in steroid receptor distribution in endometrium and myometrium were found.(ABSTRACT TRUNCATED AT 250 WORDS)     

Cellular and subcellular metabolism of progesterone by the human proliferative and secretory phase endometrium and myometrium

An investigation of the metabolism of [1,2-³H]-progesterone by human proliferative and secretory phase uterine tissue and the subcellular localization and metabolism of progesterone showed that in the endometrium and the myometrium progesterone was mainly converted to 5α-pregnane-3,20-dione and 20α-hydroxy-4-pregnen-3-one. Smaller quantities of 20α-hydroxy-5α-pregnan-3-one, 6β-hydroxy-progesterone and unidentified polar metabolites were also formed. Qualitatively this metabolism did not appear to vary significantly in the human endometrium and myometrium. However, quantitative variations between the endometrium and myometrium were apparent in both the proliferative and secretory phases of the cycle. Conversion of progesterone to 5α-pregnane-3, 20-dione was higher in the endometrium than in the myometrium, while more of 20α-hydroxy-4-pregnen-3-one was formed by the myometrium. More 20α-hydroxy-4-pregnen-3-one was formed in the secretory than in the proliferative phase.Localization of progesterone in the subcellular fractions of the endometrium and myometrium showed that progesterone and its metabolites are mainly localised in the cytosol fraction, wherein they bind specifically to receptor protein. The nuclear uptake was considerably lower than that of the cytosol fraction. The subcellular metabolism of progesterone in the individual fractions, in the presence of co-factors, revealed that the conversion of progesterone was higher in the cytosol fraction. The major metabolite formed in the nuclear fraction of endometrium and myometrium was 5α-pregnane-3,20-dione. In the mitochondrial fraction 5α-pregnane-3,20-dione, 20α-hydroxy-5α-pregnan-3-one and some highly polar compounds were formed. 6β-hydroxy-progesterone was formed to a considerable extent by the microsomal and mitochondrial fractions. In the cytosol fractions of both the endometrium and myometrium, 20α-hydroxy-4-pregnen-3-one was the major metabolite with small amounts of 6β-hydroxy-progesterone, 5α-pregnane-3, 20-dione and 20α-hydroxy-5α-pregnan-3-one.     

Changes during the menstrual cycle in cytosolic and nuclear concentrations of progestagen receptor in the human Fallopian tube

[3H]R5020 was bound to cytosolic and nuclear samples of human Fallopian tube with high affinity and specificity. The cytoplasmic and nuclear concentrations of progestagen receptor varied, throughout the menstrual cycle, in the ampulla, isthmus and fimbria. Concentrations were higher at the late proliferative stage of the cycle than at the early proliferative and late secretory stages. A positive linear regression was observed between cytosolic and nuclear progestagen receptor concentrations and plasma oestradiol levels. A negative linear relationship was observed between cytosolic progestagen receptor concentration and plasma progesterone levels during the secretory stages of the menstrual cycle.     

Effects of 17 beta-estradiol and progesterone on the levels of prostaglandins F2 alpha and E in human endometrium

17 beta-estradiol and progesterone were administered to post-menopausal women to determine their effects in vivo on the capacity of human endometrium to synthesize prostaglandins (PGs) F2 alpha and E. Basal amounts of PGF2 alpha and PGE synthesized by endometrium exposed to 17 beta-estradiol and progesterone were significantly higher than the levels produced by endometrium exposed to 17 beta-estradiol alone (p less than 0.02 for both PGs). Levels found in the former endometrium were broadly comparable to levels in secretory endometrium and in the latter to amounts found in proliferative endometrium of spontaneous, ovulatory cycles.     

Menstrual cycle-associated alteration of sulfogalactosylceramide in human uterine endometrium: possible induction of glycolipid sulfation by sex steroid hormones

The human uterine endometrium is a tissue in which cell proliferation and differentiation are strictly controlled by sex steroid hormones, and these hormone-controlled cellular events occurring in association with the menstrual cycle of the uterine endometrium should be accompanied by characteristic molecular and metabolic changes. To characterize the menstrual cycle at the molecular level, we analyzed the glycolipids of human uterine endometrium in the proliferative and secretory phases of the menstrual cycle. Neutral glycosphingolipids from uterine endometrium comprised globo-series glycosphingolipids, such as GlcCer, LacCer, Gb3Cer, and Gb4Cer, and the relative concentrations remained constant in the two phases. However, in the case of acidic glycosphingolipids, although the concentrations of sialoglycosphingolipids remained at constant levels in the two phases, sulfatide, I3-SulfoGalCer, dramatically increased from the proliferative to the secretory phase, amounting to 7-17 nmol/g dry weight in the proliferative phase and 115-245 nmol/g dry weight in the secretory phase. Since sulfatide was the only glycolipid that changed in association with the menstrual cycle, it is likely that the sulfotransferase responsible for the synthesis of sulfatide might be induced by sex steroid hormones, estrogen and progesterone, and that sulfatide might play an essential biological role in the secretory phase of the menstrual cycle in the uterine endometrium.     

In vitro evaluation of estrogenic, estrogen antagonistic and progestagenic effects of a steroidal drug (Org OD-14) and its metabolites on human endometrium

The human endometrial model for in vitro evaluation of estrogenic, estrogen antagonistic, and progestagenic effects of endogenous steroids, natural products or synthetic drugs was applied to the study of Org OD-14, an analog of norethynodrel developed by Organon International, Oss, The Netherlands, and some of its metabolites. Estrogen antagonistic actions of Org OD-14 and its 4-ene isomer were evident from their ability to suppress the enhancement of PGF2 alpha output elicited by estradiol on fragments of secretory endometrium and to decrease the rate of output of the prostaglandin by proliferative tissue, already stimulated by endogenous estrogens. These inhibitory effects were similar to those obtained with progesterone and do not appear to involve competition for the estrogen receptor since the antiestrogen 4-hydroxyamoxifen was not active in parallel incubations of proliferative endometrium. The progestagenic effects of Org OD-14 and its 4-ene isomer were also evident from their capability to enhance estradiol 17 beta-dehydrogenase activity and glycogen accumulation in specimens of proliferative endometrium. Estrogenic effects of the 3 alpha- and 3 beta-hydroxy metabolites of Org OD-14 were demonstrated by their stimulatory actions on PGF2 alpha output during incubations of secretory endometrium. The estrogenic and progestagenic actions of these compounds are in general agreement with their relative affinity for binding to the estradiol and progesterone receptors, although their actions may be influenced by intracellular metabolism in the endometrial tissue. For instance, the similarity in progestagenic activity of Org OD-14 and the 4-ene isomer, contrasting with their different affinities for the progesterone receptor, may result from in situ isomerization of Org OD-14 to the 4-ene metabolite.     

Synthesis of polypeptides by the cervix of the baboon (Papio anubis)

Administration of oestradiol to ovariectomized baboons caused the epithelium of the cervix to differentiate into tall columnar cells that were ciliated or secretory. Administration of progesterone in the presence or absence of oestradiol altered the appearance of the lining epithelium, suggesting a decrease in secretory activity. Fluorographs of media from cultures of tissue from steroid-treated animals reflected changes in polypeptide biosynthesis which correlated with the morphological observations: 6 polypeptides (Mr 88,000-37,000; pI 5.5-6.0) were observed in all treatment groups and, except for relative changes in intensity, these polypeptides were electrophoretically similar to those synthesized by the endometrium. A new group of low molecular weight polypeptides (Mr 23,000-20,000, pI greater than 8.0-5.5) and a basic protein (Mr 160,000) were synthesized and released in the oestradiol-dominated animal. These polypeptides were distinct to the cervical mucosa since they were not observed in the endometrium or oviduct. Progesterone suppressed the synthesis of the low molecular weight acidic polypeptides (Mr 23,000-20,000; pI 6.1-5.5) but maintained the synthesis of the basic polypeptides (Mr 23,000-20,000; pI greater than 8). Treatment with progesterone +/- oestradiol did not appear to induce the synthesis of any new major polypeptides in the cervical epithelium. These results suggest that oestradiol induces the synthesis of a group of cervix-specific polypeptides and progesterone antagonizes the action of oestradiol in the baboon cervix.     

Decidual prolactin production by organ cultures of human endometrium: effects of continuous and intermittent progesterone treatment

Explants of human endometrium were cultured in serum-free nutrient medium and the effects of continuous and intermittent progesterone treatment on decidual prolactin (dPRL) production compared. The synthesis of dPRL was induced in cultures of proliferative and secretory endometrium when progesterone (50 ng/ml) was added to the medium. The amount of dPRL produced by these cultures increased gradually during 41 days of continuous progesterone treatment. When progesterone was provided for only the first 14 days of a 28-day cycle, dPRL production continued to increase during the first wk of culture in the absence of exogenous hormone and then began to decline. A similar pattern was elicited during a second 28-day cycle. Explants of endometrium fixed for histologic examination after either continuous or intermittent progesterone treatment contained large "decidualized" stromal cells. These findings indicate that progesterone can induce and maintain decidualization and dPRL synthesis in organ cultures of human endometrium, dPRL production increases immediately after progesterone is withdrawn, and long-term dPRL production is not maintained in the absence or progesterone.     

Expression of vascular endothelial growth factor receptors in the human endometrium: modulation during the menstrual cycle

Angiogenesis is fundamental for human endometrial development and differentiation necessary for implantation. These vascular changes are thought to be mediated by the vascular endothelial growth factor (VEGF), whose specific receptors have not been examined in detail thus far. We conducted the present study to determine, by immunocytochemistry and computerized image analysis of the functionalis, the expression and modulation of the receptors Flk-1/KDR and Flt-1, which mediate VEGF effects on endothelial mitogenicity, chemotaxis, and capillary permeability. VEGF receptors are expressed mainly in endometrial endothelial cells, with variations of intensity and number of stained capillaries related to the phase of the cycle. The number of capillaries immunostained for Flk-1/KDR was maximal in the proliferative phase (ratio Flk-1/CD34: 1), twice as high as the number of Flt-1-expressing capillaries (ratio Flt-1/CD34: 0.47). The staining intensity for Flk-1 decreased during the late proliferative and early secretory phases, to increase again in the midsecretory period. The number of Flt-1-labeled capillaries was about 2-fold higher in the secretory than in the proliferative phase; however, the proportion of Flt-1-positive cells did not change, owing to the associated increase in vascular density that characterizes progression of the functionalis from the proliferative to the secretory stage. The staining intensity for Flt-1 was higher during the late proliferative and secretory phases (especially in the midsecretory phase) and the premenstrual period. In contrast, the proportion of capillaries expressing Flk-1/KDR decreased in the secretory phase (ratio Flk-1/Von Willebrand factor: 0.55). Enhanced expression of Flk-1/KDR, and of Flt-1, on narrow capillary strands at the beginning of and during the proliferative phase may account for the rapid capillary growth associated with endometrial regeneration following menstrual shedding. The high coexpression of Flk-1/KDR and Flt-1 observed on capillaries during the midsecretory period correlates with an increase of endometrial microvascular density and of permeability characteristic of this phase of the cycle, which is a prerequisite for implantation. Finally, strong expression of Flt-1, but not Flk-1/KDR, was observed on dilated capillaries during the premenstrual period and the late proliferative phase, suggesting preferential association of Flt-1 with nonproliferating capillaries at those times; activation of this receptor by VEGF could be involved in premenstrual vascular hyperpermeability, edema, and extravasation of leukocytes. In addition to the endothelial localization, we found that epithelial cells expressed Flt-1 and Flk-1/KDR. We conclude that Flt-1 and Flk-1/KDR in the functionalis are modulated in parallel or independently according to the phase of the cycle, and that these changes are responsible for VEGF actions on endometrial vascular growth and permeability. The molecular mechanisms concerning these regulations will require further investigation.     

Multidrug resistance gene expression is controlled by steroid hormones in the secretory epithelium of the uterus

The multidrug resistance (mdr) gene family has been shown to encode a membrane glycoprotein, termed the P-glycoprotein, which functions as a drug efflux pump with broad substrate specificity. This multigene family is expressed in a tissue-specific fashion in a wide variety of normal and neoplastic tissues. The regulation of mdr gene expression in normal tissues is not understood. We have recently shown that mdr mRNA and the P-glycoprotein increases dramatically in the secretory luminal and glandular epithelium of the gravid murine uterus. This observation has suggested that mdr gene expression in the uterus is controlled by the physiologic changes associated with pregnancy. This report now demonstrates that mdr mRNA and P-glycoprotein are induced at high levels in the uterine secretory epithelium by the combination of estrogen and progesterone, the major steroid hormones of pregnancy. This regulation of mdr gene expression in the uterus does not require any other contribution from the fetus or placenta. The data indicate that this gene locus is hormonally responsive to estrogen and progesterone in the uterine secretory epithelium, suggesting an important and physiologically regulated role during pregnancy.     

Effects of 17ß-estradiol and progesterone on the levels of prostaglandins F2α and E in human endometrium

17ß-estradiol and progesterone were administered to post-menopausal women to determine their effects on the capacity of human endometrium to synthesize prostaglandins (PGs) F₂α and E. Basal amounts of PGF₂α and PGE synthesized by endometrium exposed to 17ß-estradiol and progesterone were significantly higher than the levels produced by endometrium exposed to 17ß-estradiol alone (p < 0.02 for both PGs). Levels found in the former endometrium were broadly comparable to levels in secretory endometrium and in the latter to amounts found in proliferative endometrium of spontaneous, ovulatory cycles.     

Effects of ovariectomy and steroid replacement on the genital tract of the viviparous lizard, Xantusia vigilis

Two glandular components are described in the genital tract of Xantusia: tubal glands in the Fallopian tube and goblet cells in the uterine villi. Sperm or seminal receptacles occur between adjacent villi in the uterus. Forty ovariectomized lizards carrying a silk loop in the wall of the left uterus were treated for two weeks with either progesterone, estradiol-17 β, progesterone plus estradiol or vehicle. Uteri with loops serving as a local irritant, did not differ significantly from the contra-lateral uteri in any group, hence a response similar to the deciduomal reaction of mammals is not found in this lizard. The weight of the genital tract is similar in sham-operated and in ovariectomized lizards injected with either progesterone or the vehicle. Maximal increase in weight of the tract is noted with estradiol treatment, while simultaneous administration of both steroids is followed by a moderate increase of oviducal weight. Tubal glands and sperm receptacles in ovariectomized lizards injected with either the vehicle or progesterone are smaller than those of the sham-operated or ovariectomized lizards treated with estradiol or with estradiol plus progesterone. Goblet cells are small and lack secretory granules in ovariectomized lizards injected with either the vehicle, or with estrogen or progesterone alone. Both steroids, given together, restore the size and apparent secretory activity of the goblet cells. It is concluded that in this viviparous species, both estrogen(s) and progestin(s) are essential for the maturation of the genital tract in the preovulatory stage.     

正常妇女不同卵巢激素状态离体输卵管平滑肌自发收缩活动及其对NE反应性研究

通过离体实验方法观察58例22—40岁中国育龄正常妇女周期的不同阶段、妊娠期(1—2个月)及哺乳期(2—10个月)输卵管峡部平滑肌收缩频率、波型及对外源性去甲肾上腺素(NE)的反应。实验观察到增生期、行经期及哺乳期峡部收缩活动比分泌期、妊娠期强,前者收缩频率高,多数呈现阵发性收缩,而后者收缩频率低,呈现单个收缩。增生期、行经期、分泌期及哺乳期峡部对NE呈兴奋性反应,只有妊娠期峡部对NE呈抑制反应。峡部环层肌比纵层肌收缩力强,但收缩波型相似。结果表明人输卵管自发收缩活动及对NE反应与卵巢激素状态有密切关系。     

Regulation of bcl-2 gene family members in human endometrium by antiprogestin administration in vivo.

It is likely that the changes which occur in the endometrium throughout the menstrual cycle involve apoptosis, and that expression of associated genes, such as the bcl-2 family, are regulated by sex steroids. The aim of this study was to investigate the presence of bcl-2, Bax and oestrogen receptor proteins in secretory endometrium collected from ten patients with normal ovulatory cycles 4 or 6 days after the LH surge, and on the same days in a subsequent cycle in which the formation of secretory changes was inhibited by the administration of the antiprogestin mifepristone (RU486) 2 days after the onset of the LH surge. Since some stromal cells display positive immunoreactivity, leucocyte subpopulations of macrophages (CD68-positive) and large granular lymphocytes (CD56-positive) were identified in serial sections. After administration of mifepristone on day 2 after the LH surge, a significant increase in bcl-2 immunoreactivity was observed in glandular and surface epithelium. A positive correlation (0.874) with nuclear oestrogen receptor immunoreactivity in endometrial glands was demonstrated. Subsets of stromal cells, identified as CD68-positive macrophages and CD56-positive large granular lymphocytes displayed positive immunoreactivity for the bcl-2 epitope, which was unaffected by mifepristone administration. Bax immunostaining was similar in control and antiprogestin-treated endometrium. These data indicate that antiprogestin administration inhibits progesterone downregulation of steroid receptors in endometrial glands, resulting in persistence of a proliferative endometrium and accompanying bcl-2 secretion.