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1.
Positive and negative effects of estradiol-17 beta in the rat uterus   总被引:1,自引:0,他引:1  
Estrogens could act as effectors or inhibitors of protein synthesis in the rat uterus, depending on the doses given to animals. A single injection of estradiol-17 beta to immature female rats led to the increase in protein synthesis and in enzyme activities involved in DNA synthesis. Four injections, given once daily, resulted in the inhibition of enzyme activity and synthesis of all proteins but one. The 105 kD protein which showed a gradual increase with the duration of estrogen treatment could be responsible for the negative action of estrogens on uterine growth.  相似文献   

2.
T Ord?g  Z Vértes  M Vértes 《Life sciences》1992,51(15):1187-1196
Opioid drugs and peptides were investigated for their effect on uterine DNA synthesis induced by a single injection of 17 beta-oestradiol given to ovariectomized rats 24 h prior to decapitation. [D-Met2,Pro5]-enkephalinamide administered 12, 2 or 1 h before killing resulted in a significant (approximately 50%) inhibition of in vitro [3H]-thymidine incorporation into DNA, while injections given 24 or 6 h before decapitation were ineffective. Non-linear regression of the dose-effect curves resulted in an ED50 of approximately 0.26 and approximately 0.45 microgram/100 g b.wt. for the opioid treatments given 12 or 2 h before killing, respectively. These effects could be completely reversed by the opioid antagonist naloxone injected 30 min prior to the agonist treatment, while naloxone itself had no effect. Morphine and [D-Ala2,D-Leu5]-enkephalin administered 12 h, as well as dynorphin A fragment 1-13 given 2 h before decapitation also inhibited oestradiol-induced uterine DNA synthesis. In ovariectomized animals without 17 beta-oestradiol priming no significant effect of [D-Met2,Pro5]-enkephalinamide or naloxone on [3H]TdR incorporation was found.  相似文献   

3.
We report here results of a study of the effect of sequential administration of 1 microgram 17 beta-estradiol in vivo on the incorporation of L-[35S]methionine into specific proteins in vitro in the immature rat uterus. One-dimensional SDS-PAGE analysis of labeled secreted uterine proteins and cellular proteins extracted from the luminal epithelial and from the stroma plus myometrial uterine fractions revealed that estradiol preferentially stimulated the synthesis of 110 K, 74 K and 66 K secreted proteins, 180 K and 110 K epithelial proteins and a 175 K stroma-myometrial protein among others, while it decreased the relative rate of synthesis of a 32.5 K secreted protein and a 70 K stroma-myometrial protein. The 110 K protein, a secreted luminal epithelial protein whose labeling in vitro dramatically increased greater than 60-fold per mg endometrial DNA after in vivo estrogen stimulation, may be a useful marker for studying estrogen action in the luminal epithelium of the immature rat uterus. Comparison of the secreted proteins labeled at 28 h (4 h after a second injection) and at 54 h (6 h after a third injection) revealed that estradiol effected a sequential change in the pattern of synthesis of secreted uterine proteins in vitro. Comparison of the number and magnitude of changes in the synthesis of specific proteins in the luminal epithelium and the stroma plus myometrium revealed that protein synthesis in the luminal epithelium is clearly more responsive to estradiol and readily distinguishable from the responsiveness of the stroma plus myometrium.  相似文献   

4.
Oestrogen, progesterone and androgen inhibit uterine cell death after the depletion of oestrogen. In the present study, we investigated effects of glucocorticoid on death of mouse uterine cells. Castrated female mice were given a daily injection of 17 beta-oestradiol (0.2 microgram/mouse/day) for 3 days, and then an injection of 5'-[125I]iodo-2'-deoxyuridine ([125I]IdUrd) to label DNAs of uterine cells with 125I. Mice were killed at intervals during subsequent treatments, and the retention of [125I]IdUrd incorporated into the whole uterus was determined. On subsequent injection of vehicle only, the 125I-radioactivity retained in the whole uterus rapidly decreased. Injections of dexamethasone (50 micrograms/mouse/day) reduced the loss of 125I-radioactivity slightly but significantly. Dexamethasone also showed synergistic effects on the retention of 125I-radioactivity when it was daily injected together with 17 beta-oestradiol, progesterone or 5 alpha-dihydrotestosterone. The present results suggest that glucocorticoid may affect the processes involved in the uterine cell death, in a manner such as inhibiting the uterine cell death or delaying the removal of DNAs of dead cells from the uterus.  相似文献   

5.
Although cystic fibrosis transmembrane conductance regulator (CFTR) has been shown to be expressed in the female reproductive tract, its functional role in the uterus is not fully understood. The present study investigated a possible physiological role of CFTR by comparing the effects of 17beta-oestradiol and Bak Foong Pill (BFP), an over-the-counter Chinese medicine used for centuries for the treatment of various gynaecological disorders, on uterus size and the expression of CFTR in the uterus of ovariectomised mice using RT-PCR. Treatment of ovariectomised mice with 17beta-oestradiol (0.2 mg/kg, p.o.) for 12 days caused a significant increase in uterine wet weight compared to vehicle. However, treatment with BFP (3 g/kg, p.o.) for the same period failed to increase uterine wet weight, indicating a lack of direct oestrogen-like activity of BFP. Analysis of CFTR mRNA expression in the harvested uteri using RT-PCR showed that both 17beta-oestradiol and BFP induced an increase in CFTR mRNA expression in mouse uteri compared to levels observed in vehicle-treated animals. These results suggest that CFTR can be upregulated by oestrogen and BFP, however, the effect exerted by BFP does not seem to be mediated by direct oestrogen-like activity. Regulation of CFTR expression by both oestrogen and gynaecological medication BFP indicates an important role of CFTR in reproductive functions.  相似文献   

6.
In order to elucidate the effects of dehydroepiandrosterone sulfate (DHAS) on softening and dilatation of the uterine cervix, changes of oestriol, 17 beta-oestradiol and progesterone levels in serum and cervix, Bishop score and collagenase activity in the cervical tissue were assessed in pregnant women before and after treatment with DHAS. 17 beta-oestradiol level in the serum and cervical tissue markedly increased after the administration of DHAS, while oestriol level remained unchanged. Serum progesterone level did not change in the majority of cases, while it decreased within several hours in patients in whom delivery was accomplished within 24 hours after the administration of DHAS. Among the factors connected with the Bishop score, effacement and consistency of the cervix were remarkably improved by DHAS administration. Total collagenase activity in the cervical tissue of patients treated with DHAS was elevated by an average of 152%. These results suggest that DHAS is potent in ripening the uterine cervix through an activation of collagenase activity induced by the enhanced conversion to 17 beta-oestradiol. Thus, DHAS administration in the late stage of pregnancy is valuable in prepartal treatment for induction of labour.  相似文献   

7.
(1) Pharmacological concentrations (greater than 10(-5) M) of 17 beta-oestradiol inhibited 35S-labelled proteoglycan synthesis in bovine articular cartilage explant cultures. They also inhibited 35S-labelled proteoglycan synthesis and 3H-labelled protein synthesis in cell cultures of chondrocytes from bovine articular cartilage and Swarm rat chondrosarcoma. Maximal inhibition was about 30-50%. Physiological concentrations (10(-9)-10(-8) M) of oestradiol had no effect on the synthesis of either protein or proteoglycan. (2) The inhibitory action of high concentrations of oestradiol on these biosynthetic pathways is not common to all steroids since 10(-4) M cortisol had no effect on articular chondrocyte cell cultures. 10(-4) M testosterone had a similar action to oestradiol. (3) Neither physiological nor pharmacological concentrations of 17 beta-oestradiol had any effect on 35S-labelled proteoglycan turnover in the cartilage explant system. (4) 10(-5) M oestradiol inhibited cell division in cultures of articular chondrocytes which had entered the log growth phase. 10(-7) M oestradiol had no effect on articular chondrocyte growth. (5) In male rats implanted with silastic capsules releasing 17 beta-oestradiol, increase in body weight was retarded by about 25% over a period of 6 weeks, compared to control rats. Rat chondrosarcoma grew to the same size in oestrogen-treated rats as it did in controls. (6) Oestrogen receptors could not be detected in freshly isolated bovine articular chondrocytes or in rat chondrosarcoma. (7) In conclusion, neither the mitotic rate of articular chondrocytes nor their proteoglycan metabolism is under the direct physiological control of oestradiol. Growth and biosynthetic activity of the rat chondrosarcoma chondrocytes are independent of either direct control by the hormone or control effected by oestradiol regulation of a second hormone or growth factor.  相似文献   

8.
Immature rat uterine nucleoli were isolated and their ability to synthesize RNA in vitro was determined. Estradiol-17β injected intraperitoneally 2 hr prior to killing stimulated rat uterine nucleolar in vitro RNA synthesis both quantitatively and qualitatively. The intraperitoneal administration of cycloheximide as late as 10 min prior to the end of a 2-hr estrogen exposure prevented both the quantitative and qualitative changes stimulated by estrogen. The data suggest that estrogen-stimulated rat uterine nucleolar RNA synthesis requires the continuous synthesis of protein.  相似文献   

9.
Estrogen regulation of c-fos messenger ribonucleic acid   总被引:8,自引:0,他引:8  
Acute administration of 17 beta-estradiol to immature female rats elicits a rapid and striking increase in the size of the uterus. This increase in size to caused by both hypertrophy and hyperplasia in the epithelial, stromal, and myometrial cells in the uterus. Previous studies have shown that induction of mRNA for the epidermal growth factor receptor, the cellular homolog of the erb-B oncogene, occurs early during estrogen-stimulated uterine growth. We report here that estradiol causes a very rapid induction of the mRNA for the cellular oncogene c-fos in immature rat uterus. Steady state levels of c-fos mRNA reach a maximum 3 h after 17 beta-estradiol administration and slowly return to low basal levels in 15 h. Dexamethasone, progesterone, and 5 alpha-dihydrotestosterone had no effect on uterine c-fos mRNA expression. The induction of c-fos mRNA by estrogen was unaffected by the protein synthesis inhibitor puromycin but completely abolished by the RNA synthesis inhibitor actinomycin D.  相似文献   

10.
The changes in the activities of 17 aminoacyl-tRNA synthetases induced by phosphorylation [1] were reversed by the action of cyclic AMP in preparations from both uterus and liver. Cyclic AMP also inhibited the phosphorylation of aminoacyl-tRNA synthetase protein by endogenous non-cyclic AMP-dependent protein kinase and [gamma-32P]ATP. The effect was not due to a stimulation of phosphoaminoacyl-tRNA synthetase phosphatase or to an influence of cyclic AMP on aminoacyl-tRNA synthetases. The activity of phosphoaminoacyl-tRNA synthetase phosphatase was increased by treatment with endogenous cyclic AMP-dependent protein kinase, ATP and cyclic AMP. Affinity chromatography of the 32P-labeled phosphorylated phosphosynthetase phosphatase protein followed by gel electrophoresis showed that the activated phosphatase was phosphorylated. In the uterus, the changes in 17 aminoacyl-tRNA synthetase activities observed 5 min after dibutyryl cyclic AMP administration to ovariectomized mice were similar to those observed after 17beta-oestradiol treatment, whereas in the liver the changes in these activities were the opposite to those found after treatment with 17beta-oestradiol. A mechanism for the regulation of the 17 aminoacyl-tRNA synthetase activities is proposed, which suggests that the synthetase activities inhibited (group I) or stimulated (group II) by phosphorylation with a non-cyclic AMP-dependent aminoacyl-tRNA synthetase kinase are reactivated (group I) or inhibited (group II), respectively, by the action of a cyclic AMP-dependent phosphatase kinase through the increased activity of phosphorylated phosphoaminoacyl-tRNA synthetase phosphatase.  相似文献   

11.
This study examined the effects of heat stress on conceptus secretion of bovine trophoblastic proteins (bTP) and the uterine secretory environment on Day 17 of pregnancy. After mating to fertile bulls, cows were placed in an environmental chamber at 21 degrees C, 45% RH on Day 7 and were assigned to one of the following treatments on Day 8: Control (21 degrees C, 45% RH), Treatment 1 (37 degrees C, 30% RH) and Treatment 2 (37 degrees C, 40% RH). Cows were slaughtered on Day 17 and each uterine horn was flushed separately with physiological saline to recover the conceptus and uterine contents. The wet weight of the conceptus was recorded and uterine flushings were analyzed for quantitative and qualitative protein changes, prostaglandin F and calcium concentrations. Conceptus tissue was cultured in vitro with 100 muCi of [(3)H]-leucine and the polypeptides released into the medium were analyzed by 2D-PAGE followed by fluorography. Heat stress increased rectal temperature of cows and reduced conceptus wet weight. Uterine content of protein and calcium were increased in the uterine horn ipsilateral to the CL of heat-stressed cows. Although uterine protein increased in heat-stressed cows, no qualitative difference was observed in the polypeptides present in the uterine lumen. Conceptus synthesis and release of bTP were enhanced in treated cows. These responses indicate that heat stress between Days 8 to 17 of pregnancy altered the uterine environment as well as growth and secretory activity of the conceptus.  相似文献   

12.
In rodent uterus, both up- and down-regulation of estrogen receptor alpha (ERalpha) messenger ribonucleic acid (mRNA) and protein levels by estradiol has been demonstrated; however, it is not known which of the uterine compartments (endometrial epithelium, stroma, myometrium) respond to estradiol with autoregulation of ERalpha. The purpose of the present study was to investigate and compare the kinetics and cell type-specific effects of estradiol on uterine ERalpha expression in immature and adult rats. Ovariectomized female rats were injected s.c. with sesame oil or estradiol-17beta. Uteri were collected and analyzed for changes in ERalpha mRNA using RNase protection assays (RPA) and in situ hybridization using radiolabeled probes specific for ERalpha. Immunohistochemical analysis was performed with a polyclonal antibody specific to ERalpha. Expression of ERalpha in the uterine epithelial cells decreased at 3 and 6 h after estradiol administration to immature and adult rats, respectively. At 24 h, ERalpha mRNA levels in the immature and mature rat uterus were higher than pretreatment levels but returned to baseline by 72 h. Pretreatment with cycloheximide did not block the 3-h repressive effect of estradiol, suggesting that the estradiol-induced decrease in ERalpha mRNA occurs independent of new protein synthesis. A decrease in ERalpha mRNA and protein was also observed in uterine epithelia at 3 and 6 h after an estradiol injection to immature and adult rats, and intensity of both the in situ hybridization signal and the immunostaining in the epithelium increased at 24 and 72 h. However, the periluminal stromal cells in the adult uterus and the majority of stromal cells of the immature uterus appeared to have increased ERalpha expression. The results indicate that down-regulation of ERalpha in the epithelia and up-regulation of stromal ERalpha play a role in early events associated with estradiol-induced cell proliferation of the uterine epithelia.  相似文献   

13.
The secretion and synthesis of protein in vitro by explants of endometrium were examined in entire ewes during the first 10 days of the oestrous cycle and during an equivalent interval in ovariectomized ewes which received injections of oestradiol and progesterone. The schedule of steroid injections given was designed to simulate endogenous ovarian secretion of progesterone during the luteal phase before oestrus, of oestradiol around oestrus and of progesterone during the luteal phase after oestrus. The rate of protein synthesis and tissue RNA:DNA and protein:DNA ratios in intercaruncular and caruncular endometrium were generally higher in entire than in ovariectomized ewes. In ovariectomized ewes oestradiol increased these activities at 2-4 days after oestrus, whereas progesterone preceding oestradiol caused increases at oestrus, but not thereafter. In entire ewes and in ovariectomized ewes receiving the full steroid treatment regimen, protein secretion was high at oestrus and declined markedly during the next 4-6 days. In ovariectomized ewes not receiving progesterone before oestradiol, secretion increased between 4 and 6 days after oestrus, or during the equivalent stage of treatment in ewes which did not show oestrus. The omission of this progesterone did not modify secretion by caruncular endometrium. Oestradiol increased protein secretion by both tissues. The data suggest that progesterone given before oestradiol (or its equivalent in entire ewes) inhibits the secretion, at about 4-7 days after oestrus, of uterine proteins which may impair embryo development in ovariectomized ewes which do not receive this progesterone.  相似文献   

14.
Estrogen regulation of tissue-specific expression of complement C3   总被引:14,自引:0,他引:14  
The administration of estradiol to immature rats results in the increased synthesis and secretion of a 180-kDa protein, composed of 115- and 65-kDa subunits, by the uterine luminal epithelial cells. A monoclonal antibody against the 180-kDa protein was utilized to isolate the corresponding cDNA (LE-1) from a rat uterine luminal epithelial cell cDNA lambda gt11 expression library. This LE-1 cDNA was sequenced and shown to be homologous to complement component C3. The sequence was approximately 81 and 90% homologous to human and mouse C3, respectively. The LE-1 cDNA sequence was homologous with the 3' portion of the C3 mRNA containing the alpha subunit (115 kDa). Uterine mRNA isolated from immature rats treated with 1 microgram of estradiol for 24 h demonstrated a 25-fold increase in the concentration of a 6.0-kilobase mRNA by Northern hybridization with either LE-1 or authentic human C3 cDNA probes. To further examine the possibility that the estradiol-regulated secretory protein was C3, an aliquot of radiolabeled media protein from control and estradiol-stimulated rat uteri was incubated with goat anti-rat C3 antibody. The immunoprecipitated radiolabeled protein from estradiol-treated animals was increased significantly (p less than 0.01) compared to media from control animals. Analysis of the immunoprecipitated proteins on nonreducing sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed a single protein of 180 kDa from estradiol-stimulated uterine media, whereas no detectable proteins were immunoprecipitated from media obtained from control uteri. Also, when the immunoprecipitated protein was reduced (20 mM dithiothreitol) it dissociated into two subunits of 115 and 65 kDa. Immunohistochemical studies demonstrated the presence of C3 only in the epithelial cells of estrogen-stimulated rat uteri. In addition, the estradiol-stimulated mRNA was only detectable in uterine epithelial cell RNA. Analysis of liver RNA demonstrated a 6.0-kilobase mRNA, as in the uterus, when hybridized with LE-1. However, unlike the uterus, its concentration was not influenced by estrogen administration with up to three daily injections of 100 micrograms of diethylstilbestrol. Based on biophysical, DNA sequence, and antibody data we conclude that rat uterine epithelial cells produce C3 in response to estradiol whereas the expression in the liver was not modulated by estrogens.  相似文献   

15.
We have studied the synthesis of nuclear matrix proteins as it occurs in the rat liver. To investigate their kinetics in tissue, nuclear matrix proteins were prepared from liver of rats injected with radioactive methionine. Synthesis of lamins was not observed in quiescent hepatocytes although they were the principal proteins of this subcellular fraction, suggesting that lamins are very stable in the liver. When hepatocytes were stimulated to divide by partial hepatectomy, only synthesis of lamin B was initiated. Many proteins not visible on Coomassie blue-stained gels were detectable by autoradiography. In the nuclear matrix extracts of quiescent hepatocytes, one of the most prominently labeled ones was a protein of 70 kDa. After hepatectomy, an additional protein of 62 kDa was detectable. These proteins were visible 1 h after the injection of radioactivity, but were no longer observed in nuclear matrices prepared 24 h after injection. These experiments indicate that in addition to lamins, two nuclear matrix proteins are present in the rat liver that were not detected previously, perhaps because of their rapid turnover.  相似文献   

16.
Uterine tissue isolated from immature rats at different times after estradiol injection was incubated with medium containing [3H]lysine. The acid-extractable protein from the uterine tissue was subjected to electrophoresis on sodium dodecyl sulfate and acid-urea-Triton X-100 polyacrylamide gels, and the rate of chromatin protein synthesis determined by densitometric analysis of the fluorographs of the gels. Synthesis of chromatin proteins (histones and high mobility group chromatin proteins) was stimulated by 3 h after estrogen treatment and reached a peak at 9 h, several hours before DNA synthesis was stimulated. Synthesis of chromatin proteins occurred at the same time as total cellular protein synthesis. Estrogen stimulated the synthesis of histone variants at different rates, but the accumulation of histone proteins remained coordinated such that equivalent amounts of histone proteins were being produced at any one time.  相似文献   

17.
Restriction of the conceptus to one uterine horn of the pregnant ewe results in the accumulation of fluid called uterine milk (UTM) in the contralateral horn. Two basic polypeptides, called the uterine milk proteins (UTM-proteins; Mr = 55,000 and 57,000 as determined by polyacrylamide-gel electrophoresis using sodium dodecyl sulfate), accounted for the majority of the protein in uterine milk. The two UTM-proteins were glycoproteins and were readily purified from uterine fluids by cation-exchange chromatography on carboxymethyl (CM)-cellulose followed by Sephacryl S-200 gel-filtration. The purified UIM-proteins had a weight-average molecular weight of 50,700 +/- 4,200, as determined by equilibrium sedimentation analysis. Endometrial explants from pregnant ewes were cultured in the presence of radioactive amino acids and released UTM-proteins into the medium as their major secretory products. The UTM-proteins were secreted into the uterine lumen of nonpregnant, ovariectomized ewes given daily injections of progesterone. Estrone alone was ineffective in inducing UTM-protein production. Immunocytochemical studies indicated that synthesis of the UTM-proteins was confined to the surface and glandular epithelium of the uterus.  相似文献   

18.
Rat physiologic urinary proteins were immunohistochemically localized. In male Wistar rats, urinary protein antigens were present in both hepatic and epithelial cells of the salivary ducts, the coagulating gland, and the prostate gland. In female rats, urinary protein antigens were present in the same proportion of the salivary glands as in males and in the uterine glands, but to a lesser extent than in salivary glands. The results of this study indicate multiple origins of rat urinary proteins. It remains to be determined if female uterine glands contribute to urinary proteins.  相似文献   

19.
Intraperitoneal administration of alpha-zearalenone (ZEN) to female immature rats induced synthesis of a uterine protein which has been identified as creatine kinase. The induced enzyme was purified to homogeneity by chromatography on DEAE Sephacel and Hydroxyapatite Ultrogel. Guinea pig antiserum against estrogen-induced uterine, rat uterine creatine kinase crossreacted with the ZEN-induced enzyme, indicating that ZEN exhibits an early estrogenic response in a manner analogous to natural estrogens.  相似文献   

20.
The synthesis of uterine-soluble proteins during early pregnancy in the rat has been examined by means of dual-isotope labelling techniques and subsequent electrophoretic analysis. A protein of similar electrophoretic mobility to the uterine oestrogen-induced protein was observed, and synthesis of this 'presumptive induced protein' was maximal on Day 4 and Day 6 of pregnancy but low on day 5. Pregnancy associated protein synthesis was observed in many regions on polyacrylamide gels, including the beta-lipoprotein, alpha2-macroglobulin, post-transferrin and albumin regions. Synthesis of the post-transferrin species rapidly increased from Day 4 to reach a maximum on Day 6 in the implantation tissue. The temporal pattern of synthesis of post-transferrin protein and and of 'presumptive induced region' suggests involvement in the processes of cell proliferation and decidualization.  相似文献   

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