Animals ; Cell Hypoxia ; Glucose ; metabolism ; Mifepristone ; pharmacology ; Oxidative Stress ; PC12 Cells ; Progesterone ; pharmacology ; Rats

OBJECTIVETo explore effects of acupuncture on embryo implantation and development in the rat with dysfunctional embryo implantation.

METHODSThe pregnant rats were randomly divided into a control group, a model group and an acupuncture group. In the model group and the acupuncture group, the dysfunctional embryo implantation were developed by Mifepristone, and the acupuncture group were treated with acupuncture for one consecutive week fro the first day of pregnancy. The pregnant rate, the average embryo implantation number, weight of the uterus, ovary and the size and weight of single embryo were compared among the 3 groups, and optical microscope was used for comparison of the morphological structures of the endometrium and ovary were compared among the 3 groups.

RESULTSIn the acupuncture group, the pregnant rate, the average embryo implantation number were more significantly increased as compared with the model group (P < 0.01); in the model group, the weight of the uterus and ovary were lighter than those in the control group and the acupuncture group, with poor development of the endometrium in the model group, but development of the endometrium in the acupuncture group was similar to that in the control group.

CONCLUSIONAcupuncture can reverse Mifepristone's anti-implantation effect to a certain extent, and promote implantation and development of embryo in the rat.

Acupuncture Therapy ; Animals ; Embryo Implantation ; drug effects ; Embryonic Development ; Female ; Mifepristone ; pharmacology ; Pregnancy ; Rats

OBJECTIVEObserving the effect of phenolic acids from Arnebia euchroma assist mifepristone in anti-early pregnancy of SD rattus norvegicus.

METHODFeed the SD rattus norvegicus with phenolic acids from A. euchroma during the 7 th to 9 th day, and then we observe the restaining rate of pregnancy. At the same time, we determine the progesterone level in blood serum in the ways of radioimmunoassay.

RESULT720 g x kg(-1) enolic aids from A. euchroma can markedly increase the restaining rate of pregnancy (P < 0.05) than that only mifepristone dose (8.0 g x kg(-1)). In addition, the number of everage still bith increase, however, to the pogesterone level in blood serum. It has little effect.

CONCLUSIONThe effect of phenolic acids from A. euchroma assist mifepristone in anti-early pregnancy of SD rattus norvegicus is clear, and it dosen't work in the ways of decreasing the pogesterone level.

Abortifacient Agents ; chemistry ; pharmacology ; Animals ; Boraginaceae ; chemistry ; Female ; Hydroxybenzoates ; chemistry ; pharmacology ; Male ; Mifepristone ; pharmacology ; Pregnancy ; drug effects ; Progesterone ; blood ; Radioimmunoassay ; Rats ; Rats, Sprague-Dawley

Primate placentation involves a series of cell proliferation, immigration and apoptosis which account for the progressive tissue remodelling at the implantation site. p53 is an important proto-oncogene involved in the regulation of cell-cycle and apoptosis. To study the effect of RU486 on apoptosis and expression of p53 at the fetal-maternal interface, the location of apoptotic cells and expression of p53 were examined using in situ 3'-end labeling method, immunohistochemistry and Western blot assay at the fetal-maternal interface of normal and RU486 treated rhesus monkey. Western blot analysis showed the specificity of the anti-human antibody used with the monkey tissue. In the placental villi, the apoptotic nuclei were observed mainly in the syncytiotrophoblast and part of the cytotrophoblast within the cell column; p53 protein was detected mainly in the cytotrophoblast. In the endometrium, positive signals for apoptosis and p53 were detected in some stromal cells. After two days of mifepristone treatment, the apoptotic cells increased significantly in both placental villi and endometrium. In the villi, the increased apoptotic nuclei were mainly localized to the cytotrophoblast. At the same time, p53-positive nuclei also increased in both villous cytotrophoblast cells and endometrial stromal cells, after the treatment of RU486. These results suggest that apoptosis and expression of p53 are essential in regulating trophoblastic homeostasis by controlling its proliferation in normal placenta, whereas the up-regulation of p53 protein may play an important role in apoptosis that happens at the fetal-maternal interface induced by RU486.
Abortifacient Agents, Steroidal ; pharmacology ; Animals ; Apoptosis ; drug effects ; Chorionic Villi ; pathology ; Female ; Macaca mulatta ; Mifepristone ; pharmacology ; Placentation ; drug effects ; physiology ; Pregnancy ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics

OBJECTIVETo observe the effects of Dexamethasone (Dex), a synthetic glucocorticoid, on glucocorticoid receptor expression in the human ovarian carcinoma cell line 3AO. The molecular mechanism of glucocorticoid (GC) on 3AO cells was also studied.

METHODSThe expression and regulation of the glucocorticoid receptor in 3AO cells was studied by utilizing radioligand binding assay and quantitative RT-PCR.

RESULTSHigh affinity and low capacity GR existed in 3AO cells, in addition, GR binding activity was down-regulated by Dex in a time-dependent manner, to a level about 28.34% of control following 24 hours treatment, with a concomitant decrease in GR mRNA. The induction of alkaline phosphatase (AKP) activity by Dex was reversed by RU486, a potent glucocorticoid antagonist.

CONCLUSIONIn 3AO cells, functional GR which can be down-regulated by Dex at the protein and mRNA level, exists suggesting that the regulating effects of Dex on GR occur at least partially at the GR mRNA level, and that the cellular effects of Dex on 3AO cells might be mediated by GR.

Dexamethasone ; pharmacology ; Dose-Response Relationship, Drug ; Female ; Humans ; Mifepristone ; pharmacology ; Ovarian Neoplasms ; chemistry ; drug therapy ; RNA, Messenger ; analysis ; Receptors, Glucocorticoid ; analysis ; drug effects ; genetics ; Tumor Cells, Cultured

OBJECTIVE: To explore the reversal effect of mifepristone(MIF) on adriamycin(ADM) resistance in human breast cell line MCF-7/ADM in vitro and in vivo. METHODS: The transplantable models of MCF-7 cells resisting against adriamycin were established in nude mice by subcutaneous implantation to observe the reversal effect of MIF in vivo. The mice were randomly divided into 4 groups: a control group(treated with saline water 0.2 mL intraperitoneally and edible oil 0.5 mL orally), an MIF group (treated with mifepristone 30 mg/kg orally and saline water 0.2 mL intraperitoneally), an ADM group (treated with adriamycin 5 mg/kg intraperitoneally and edible oil 0.5 mL orally) and an ADM+MIF group (treated with ADM 5mg/kg intraperitoneally and mifepristone 30 mg/kg orally every 3 days). Tumor changes were investigated after different drug treatments. The reversal effect of 5 micromol/L MIF in vitro on the ADM resistance cell line MCF-7/ADM and non ADM resistance cell line MCF-7 was determined by 4,5-dimethylthiazol-2-yl (MTT) assay. RESULTS: (1) The inhibitory rate of 5 micromol/L of MIF for both cell lines MCF-7 and MCF-7/ADM was less than 5%, and it had no statistical difference compared with the group that was not treated with MIF(P > 0.05). (2) ADM could inhibit the growth of both MCF-7 and MCF-7/ADM,but the inhibition concentration 50 (IC(50)) of MCF-7 (0.42 mg/L) was obviously less than that of MCF-7/ADM(17.21 mg/L) (P < 0.05). (3) IC(50) of MCF-7/ADM of MIF+ADM group was 1.96 mg/L in vitro, which was significantly less than that in ADM alone group(17.21 mg/L) (P < 0.05), and 5 micromol/L of MIF reversed ADM resistance with fold-reversal of 8.78. (4) MIF had some effect on the inhibition of MCF-7/ADM cell growth in vivo, the xenograft volume in the MIF+ADM group [(232.5149 +/- 309.2377) mm(3)] was significantly smaller than that in the control group[(962.2309 +/- 261.1313) mm(3) ] after the 4 week treatment(P<0.05), and also smaller than that in the MIF group [(778.2846 +/- 42.6919) mm(3)] and in the ADM group [(508.9648 +/- 16.2609) mm(3)](P < 0.05). There was significant inhibition on xenograft weight after MIF combined with ADM treatment in vivo, and the inhibitory rate was 78.0%. CONCLUSION MIF can effectively reverse ADM resistance in human breast cancer cell line MCF-7/ADM both in vitro and in vivo.
Animals ; Breast Neoplasms ; pathology ; Cell Line, Tumor ; Doxorubicin ; pharmacology ; Drug Resistance, Neoplasm ; drug effects ; Female ; Humans ; Mice ; Mice, Nude ; Mifepristone ; pharmacology ; Neoplasm Transplantation ; Random Allocation

Animals ; Conditioning (Psychology) ; drug effects ; Extinction, Psychological ; drug effects ; physiology ; Fear ; drug effects ; physiology ; Male ; Mifepristone ; pharmacology ; Rats ; Rats, Sprague-Dawley

The regulation of a target gene expression is very important in gene therapy. However, constitutive or inappropriate expression of the genes with traditional expression system may interfere with the effect of the gene therapy, even may lead to lethal side effect. We constructed an RU486 inducible eukaryotic vector carrying DsRed protein and evaluated its regulatable effect in vitro. The single vector named PDC-RURED was constructed with molecular biological methods, which contained DsRed gene, promoter and RU486-inducible system. To minimize any potential interference, we spaced the two transcriptional elements with a 1.6 kb insulator. The vector was identified by different enzyme restrictions, sequencing analysis and PCR assay. We demonstrated the regulatable expression of this vector after transfection in HEK293 cells by fluorescence microscopy and flow cytometry. In the absence of RU486, no significant DsRed protein activation was observed, whereas in the presence of RU486 up to 40 fold activation of the DsRed protein was observed in cultured cells. The data show that the novel eukaryotic expression plasmid vector can be used to regulate the expression level of genes of interest in appropriate time under the control of RU486. This inducible expression vector provides a powerful tool for the research of gene regulation and gene therapy.
Cell Line ; Fluorescent Dyes ; metabolism ; Genetic Therapy ; methods ; Genetic Vectors ; genetics ; Humans ; Kidney ; cytology ; Luminescent Proteins ; biosynthesis ; genetics ; Mifepristone ; pharmacology ; Promoter Regions, Genetic ; genetics ; Transfection

OBJECTIVETo study the effects of glucocorticoid on skeletal muscle protein metabolism in burn sepsis and its possible mechanism.

METHODSThe rats were randomly divided into four groups with 15 rats in each group. Group B, 30% TBSA full-thickness burn was produced on the back and endotoxin (6 mg/kg bw) was given intraperitoneally after the injury to simulate burn sepsis. Groups C and D, glucocorticoid receptor antagonist RU38486 (10 mg/kg bw) was given by gavage 2 hours before or 2 hours after burn with endotoxin, respectively. Group A, the rats received only normal saline in same volume as endotoxin. Plasma levels of cortisol were determined with standard procedure. Extensor digitorium longus muscles (EDL) were procured from both legs 12 hours after the injury. After weighing, the proteolytic rate was determined in vitro in an incubation system with oxygen rich environment by high performance liquid chromatography. The gene expressions of ubiquitin, E(2)-14kDa and C2 in the muscles were determined by Northern blot analysis.

RESULTSThe weight of EDL was significantly lower in group B than in group A (t = 9.03, P < 0.01). Although the weight of EDL muscles was also lower in groups C and D than in group A, it was significantly higher than in group B (t = 2.26, 6.42, P < 0.05 or P < 0.01). The concentrations of plasma cortisol were markedly higher in groups B, C and D than in group A (t = 9.03 - 22.94, P < 0.01). A 58.8% (210/357) of the total and 335.5% (4.16/1.24) of myofibrillar proteolytic rate in group B was higher than in group A (t = 36.99 and t = 46.19, P < 0.01), respectively. The total and myofibrillar proteolytic rate in group D was 28.3% (161/567) and 49.6% (2.68/5.40) and in group C 18.9% (108/567) and 23.2% (1.25/5.40), which were lower than those in group B (t = 5.34 approximately 34.68, P < 0.01), respectively. Although the expressions of ubiquitin mRNA (2.4 kb), E(2)-14 kDa mRNA (1.2 kb) and C2 mRNA in groups C and D were significantly higher than in group A, all the values were lower than those in group B (t = 3.22, 11.32, P < 0.01), especially in group C.

CONCLUSIONSThe proteolytic rate of skeletal muscle, especially the myofibrillar proteolytic rate, was enhanced during burn with sepsis. Hypersecretion of glucocorticoid could upgrade the gene expression of ubiquitin system, resulting in hyperdegradation of skeletal muscle protein during burn with sepsis. Glucocorticoid receptor antagonist RU38486 could decrease the hyperdegradation of skeletal muscle during burn with sepsis.

Animals ; Burns ; metabolism ; Gene Expression Regulation ; Glucocorticoids ; physiology ; Hydrocortisone ; blood ; Male ; Mifepristone ; pharmacology ; Muscle Proteins ; metabolism ; Muscle, Skeletal ; metabolism ; Rats ; Rats, Wistar ; Sepsis ; metabolism ; Ubiquitin ; metabolism

OBJECTIVETo construct an inducible eukaryotic vector carrying red fluorescent protein (DsRed) and evaluate the regulation of DsRed gene expression in vitro.

METHODSThe vector pRS17-RUDsRed containing DsRed gene, promoter and RU486-inducible system was constructed using molecular biological methods. To minimize potential interference, the two transcriptional elements were spaced with a 1.6 kb insulator. Fluorescence microscopy and flow cytometry were used to observe the activation of this regulatable vector after transfection in MFC cells.

RESULTSThe vector was identified by digestion with different restriction enzymes, sequencing and PCR. In the absence of RU486, the cells transfected with the vector exhibited very low DsRed protein expression, and the addition of RU486 induced efficient DsRed expression in the cells.

CONCLUSIONThe RU486-inducible eukaryotic vector carrying DsRed protein allows effective regulation of the target gene expression in vitro, which provides a useful tool for gene regulation and gene therapy studies.

Gene Expression Regulation ; Genetic Therapy ; methods ; Genetic Vectors ; genetics ; Humans ; Luminescent Proteins ; genetics ; metabolism ; Mifepristone ; pharmacology ; Promoter Regions, Genetic ; genetics ; Stomach Neoplasms ; genetics ; pathology ; Tumor Cells, Cultured