OBJECTIVETo observe the effects of low-dose exogenous estrogen nonylphenol (NP) on the proliferation of human prostate cancer cell lines DU-145 and the expression of the membrane estrogen receptor GPR30 in the DU-145 cells.

METHODSWe exposed DU-145 cells to different concentrations of NP for 24 hours, followed by measurement of the half maximal inhibitory concentration (IC50) of the cells by cell proliferation assay and determination of the concentration of exposure to low-dose NP. We also observed the expressions of 3 estrogen receptors (ER), including ER-alpha, ER-beta and membrane estrogen receptor GPR30, in the DU-145 cells exposed to low-dose NP by RT-PCR.

RESULTSCell proliferation assay showed that within a certain range of doses, NP inhibited the proliferation of the DU-145 cells with an IC50 of 46 micromol/L, a much lower dose of NP than IC50, 0.01, 0.1.1 micromol/l NP, that can promote the proliferation of DU-145 cells. The results of RT-PCR indicated that the expressions of the three ERs in the DU-145 cells were similar to those in prostate epithelial cells, and that low-dose NP promoted the expression of GPR30.

CONCLUSIONMembrane estrogen receptor GPR30 may play a role in low-dose NP promoting the proliferation of DU-145 cells.

Cell Line, Tumor ; Cell Proliferation ; drug effects ; physiology ; Estrogen Receptor alpha ; metabolism ; Estrogen Receptor beta ; metabolism ; Estrogens ; Humans ; Male ; Phenols ; administration & dosage ; pharmacology ; Prostatic Neoplasms ; metabolism ; pathology ; Receptors, Estrogen ; metabolism ; Receptors, G-Protein-Coupled ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo observe the effects of Bushen Zhuyun Recipe (BZR) on protein expressions of estrogen receptor (ER), progesterone receptor (PR) and integrin alpha5 and beta3 in endometrium of rats at the implantation stage, for exploring the possible mechanism of the recipe in treating luteal phase defect (LPD) infertility.

METHODSFemale SD rats were randomly divided into 6 groups, the blank group, the model group, the WM group treated by Western medicine, and the three BZR groups treated by low-, middle- and high-dose BZR respectively. Rats were made to pregnancy and sacrificed at the implantation stage, their middle segment of uterus, about 1 cm in length was gotten for detecting the protein expressions by Western blot. Results The protein expressions of endometrial ER and PR were significantly higher, while those of integrin alpha5 and beta3 were significantly lower than those of the control group (P < 0.05). The protein expressions of endometrial ER and PR were significantly lower, but those of integrin alpha5 and integrin beta3 were higher in rats treated by middle- and high- dose BZR than those in model rats (P < 0.05).

CONCLUSIONBZR can raise the receptivity of rats' endometrium through down-regulating the expressions of ER, PR and increasing the protein expression of integrin alpha5 and beta3 in endometrium and thus to enhance the pregnant rate.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Embryo Implantation ; drug effects ; physiology ; Endometrium ; drug effects ; metabolism ; Female ; Integrin alphaV ; metabolism ; Integrin beta3 ; metabolism ; Pregnancy ; Rats ; Rats, Sprague-Dawley ; Receptors, Estrogen ; metabolism ; Receptors, Progesterone ; metabolism

OBJECTIVETo discuss the relationship between estradiol and the mitogenic activated protein kinase signal transduction pathway and the expression of the MAPK in the MCF-7 breast cancer cell-line.

METHODSEpithelial growth factor (EGF) and different concentration of estradiol to induce the expression of phosphospecific ERK1/2 (pERK1/2) in MCF-7 cell line was used and the expression of pERK1/2 with western-blotting was detected. Then antiestrogen ICI 182780 and MAPK inhibitor PD98059 to inhibit the expression of pERK1/2 was used. The cell cycle of MCF-7 was detected by FACS.

RESULTSEGF could significantly induce the expression of pERK1/2. Estradiol could also induce the expression of pERK1/2, but the intensity was less than the induction of EGF. The percentage of cells in the G(2)/M cell cycle after estradiol induction increased (18.38%) compared to the control group (10.52%) (P < 0.05).

CONCLUSIONSMAPK is an important regulatory signal in breast cancer. Its measurement in breast cancer tissues provides information about the degree of activation of various growth factor pathways. This molecule may also provide a molecular target for compounds designed to block cell proliferation.

Breast Neoplasms ; enzymology ; pathology ; physiopathology ; Cell Proliferation ; drug effects ; Estradiol ; pharmacology ; Female ; Humans ; MAP Kinase Signaling System ; drug effects ; physiology ; Mitogen-Activated Protein Kinases ; drug effects ; metabolism ; Receptors, Estrogen ; metabolism ; Signal Transduction ; Tumor Cells, Cultured

AIMTo investigate the effect of androgen on the proliferation, differentiation and regression of canine prostatic stromal cells in vivo and human stromal cells in vitro.

METHODSTwenty-two dogs, including 15 normal prostate dogs and 7 prostatic hyperplasia dogs, had their serum concentration of testosterone and estrodiol determined by radioimmunoassay before and after castration. The expression of androgen receptor (AR) and estrogen receptor (ER) in the prostate were analysed by immunohistochemistry and semi-quantitative RT-PCR before and after castration. Light microscopy, transmission electron microscopy and TUNEL assay were carried out successively before and after castration to evaluate the prostatic histomorphology. In vitro serum-free cell cultures from human prostatic stroma were established and exposed to dihydrotestosterone (DHT). The proliferation of the cell culture was detected by MTT assay. The expression of TGFbgr, bFGF, AR, and smooth muscle cell (SMC) specific proteins (myosin and/or smoothelin) were detected using immunohistochemistry and RT-PCR. The differentiation from fibroblasts to smooth muscle cells was deduced by measuring the expression of SMC specific proteins.

RESULTSBefore castration, the serum concentrations of testosterone and estrodiol were not statistically different between normal and hyperplasia groups. Following castration, the serum concentration of testosterone decreased rapidly in 2 days, and the concentration of estrodiol had no significant change compared with the pre-castration data. In the prostate, AR was presented in both the epithelial and stromal cells and the AR mRNA level was higher in hyperplasia than in normal prostate tissues (P<0.05). While ER predominantly existed in the prostate stromal cells and the ER mRNA had no difference between the hyperplasia and the normal group. Within the early phase of castration (

CONCLUSIONThe whole prostate gland is an androgen-sensitive organ with both the epithelium and stroma under the control of androgen. Androgen may direct the proliferation, differentiation and regression of stromal cells by regulating the expression of TGFbgr, bFGF, AR and smooth muscle cell specific proteins.

Animals ; Biomarkers ; Cell Differentiation ; drug effects ; physiology ; Cell Division ; drug effects ; physiology ; Cells, Cultured ; Dihydrotestosterone ; pharmacology ; Dogs ; Estradiol ; blood ; Fibroblast Growth Factor 2 ; genetics ; pharmacology ; Gene Expression ; Humans ; Male ; Muscle, Smooth ; cytology ; physiology ; Orchiectomy ; Prostate ; cytology ; physiology ; Prostatic Hyperplasia ; physiopathology ; RNA, Messenger ; analysis ; Receptors, Androgen ; genetics ; Receptors, Estrogen ; genetics ; Stromal Cells ; cytology ; physiology ; Testosterone ; blood ; Transforming Growth Factor beta ; genetics ; pharmacology

To evaluate the estrogenic activities of several chemicals such as 17 beta-estradiol (E2), rho-nonylphenol, bisphenol A, butylparaben, and combinations of these chemicals, we used recombinant yeasts containing the human estrogen receptor [Saccharomyces cerevisiae ER + LYS 8127]. We evaluated E2 was most active in the recombinant yeast assay, followed by rho-nonylphenol, bisphenol A, butylparaben. The combinations of some concentrations of 17-estradiol as a strong estrogen and bisphenol A or butylparaben as a weak estrogen showed additive estrogenic effects. Also, the combinations of some concentrations of nonlyphenol and butylparaben and combination of butylparaben and bisphenol A showed additive effects in the estrogenic activity. Therefore, the estrogenic activities of the combinations of two chemicals were additive, not synergistic.
Cloning, Molecular ; Estradiol/pharmacology ; Estrogens/classification/*pharmacology ; Estrogens, Non-Steroidal/*pharmacology ; Humans ; Kinetics ; Parabens/pharmacology ; Phenols/pharmacology ; Receptors, Estrogen/drug effects/*physiology ; Recombinant Proteins/drug effects/metabolism ; Saccharomyces cerevisiae/genetics

OBJECTIVETo explore gene expression of estrogen receptor (ERalpha, ERbeta) and androgen receptor (AR) in genioglossal muscle (GG) of adult male rats, and to investigate the effects of sex hormones on GG activities, ERalpha, ERbeta and AR expression.

METHODSGG samples were collected from 10 healthy adult male rats. Total RNA were extracted and subjected to fluorescent quantitative reverse transcription-polymerase chain reaction (FQ RT-PCR) for quantitative measurement of ERalpha, ERbeta and AR mRNAs. The other 24 male rats were randomly divided into 3 groups: control group, estrogen group (intramuscular injection of estrogen 0.1 mg/kg, twice a week) and androgen group (intramuscular injection of androgen 2.5 mg/kg, twice a week). The electromyographic activities (EMG) and contract tension of GG were investigated after 4-week treatment. The expression of ERalpha, ERbeta and AR was assessed by Western blot.

RESULTSThe mRNA expression ratios of AR/GAPDH, ERalpha/GAPDH, ERbeta/GAPDH and ERalpha/ERbeta were (295.80 +/- 127.20), (2042.00 +/- 921.57), (65.96 +/- 29.57) and (36.83 +/- 19.66), respectively. The mRNA level of ERalpha was significantly higher than that of ERbeta (P < 0.01). Compared with the control group, the EMG of GG was intensified in the estrogen group (P < 0.01). GG contractility did not change significantly (P > 0.05), and ERalpha expression in GG was up-regulated by estrogen (P < 0.05); while in the androgen group, the EMG of GG was weakened (P < 0.05). P(t) and P(0) were slightly increased (P > 0.05) and the decline rate of P(0) was markedly quickened (P < 0.05). AR and ERbeta expressions were down-regulated by androgen (P < 0.05).

CONCLUSIONSBoth AR and ER were expressed in GG of adult male rats, and ERalpha was expressed more abundantly than ERbeta. Estrogen could greatly improve activities of GG and stimulate the expression of ERalpha. Whereas, androgen could restrain activities of GG, impair its fatigue resistance capacity and inhibit the expression of AR and ERbeta.

Animals ; Estradiol ; analogs & derivatives ; pharmacology ; Gene Expression Regulation ; Male ; Pharyngeal Muscles ; drug effects ; metabolism ; physiology ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Androgen ; genetics ; metabolism ; Receptors, Estrogen ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Testosterone Propionate ; pharmacology ; Tongue ; drug effects ; metabolism ; physiology

OBJECTIVETo investigate the effects of Jingqianshu granule (JQS), a Chinese medicinal formula on the expression levels of ERa and ERbeta mRNA in hypothalamus and hippocampus of PMS model rats with liver-qi depression.

METHODThe female SD rats were divided randomly into three groups: control, model and model plus Jingqianshu granule. We make the model rats of PMS with liver-qi depression using the method of emotional stimulation multiple factors. Then we applied semi-quantitative RT-PCR gene amplification technology to detect the expression levels of ERalpha and ERbeta mRNA in the brain regions of all rats.

RESULTCompared with the control group, the expression levels of ERalpha and ERbeta mRNA in hypothalamus and hippocampus of model rats were higher significantly (P < 0.05), while in the JQS-administration group, the expression levels of ERalpha and ERbeta mRNA were lower than model group (P < 0.05), and were no difference with the control group.

CONCLUSIONThe JQS granule can down-regulate the expression levels of ERalpha and ERbeta mRNA in hypothalamus and hippocampus, which maybe one of the mechanism to treat PMS with Liver-qi depression.

Animals ; Disease Models, Animal ; Dosage Forms ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Gene Expression ; drug effects ; Hippocampus ; drug effects ; metabolism ; Humans ; Hypothalamus ; drug effects ; metabolism ; Liver ; drug effects ; physiology ; Premenstrual Syndrome ; drug therapy ; genetics ; metabolism ; Qi ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Estrogen ; genetics ; metabolism

Bisphenol A (BPA), a ubiquitous environmental contaminant, has been shown to cause developmental toxicity and carcinogenic effects. BPA may have physiological activity through estrogen receptor (ER) -alpha and -beta, which are expressed in the central nervous system. We previously found that exposure of BPA to immature mice resulted in behavioral alternation, suggesting that overexposure of BPA could be neurotoxic. In this study, we further investigated the molecular neurotoxic mechanisms of BPA. BPA increased vulnerability (decrease of cell viability and differentiation, and increase of apoptotic cell death) of undifferentiated PC12 cells and cortical neuronal cells isolated from gestation 18 day rat embryos in a concentration-dependent manner (more than 50 micrometer). The ER antagonists, ICI 182,780, and tamoxifen, did not block these effects. The cell vulnerability against BPA was not significantly different in the PC12 cells overexpressing ER-alpha and ER-beta compared with PC12 cells expressing vector alone. In addition, there was no difference observed between BPA and 17-beta estradiol, a well-known agonist of ER receptor in the induction of neurotoxic responses. Further study of the mechanism showed that BPA significantly activated extracellular signal-regulated kinase (ERK) but inhibited anti-apoptotic nuclear factor kappa B (NF-kappaB) activation. In addition, ERK-specific inhibitor, PD 98,059, reversed BPA-induced cell death and restored NF-kappaB activity. This study demonstrated that exposure to BPA can cause neuronal cell death which may eventually be related with behavioral alternation in vivo. However, this neurotoxic effect may not be directly mediated through an ER receptor, as an ERK/NF-kappaB pathway may be more closely involved in BPA-induced neuronal toxicity.
Animals ; Apoptosis/drug effects ; Blotting, Western ; Cell Differentiation/drug effects ; Cell Survival/drug effects ; Estradiol/analogs & derivatives/pharmacology ; Estrogens, Non-Steroidal/*toxicity ; Flavonoids/pharmacology ; NF-kappa B/metabolism ; Neurons/*drug effects/physiology ; PC12 Cells ; Phenols/*toxicity ; Rats ; Receptors, Estrogen/metabolism ; Tamoxifen/pharmacology

OBJECTIVETo compare the therapeutic effect of Compound Recipe Gengniankang ( GNK) with that of hormone replacement treatment (HRT) on climacteric female rats with osteoporosis, and to investigate the roles of estrogen and estrogen receptors in the mechanism of osteoporosis.

METHODSClimacteric female rats with osteoporosis were chosen and divided into three groups (GNK group, HRT group and control group). Apoptosis of ovarian granulose cells was measured by terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) assay. Serum level of estradiol (E(2)), follicle stimulating hormone (FSH), luteinizing hormone (LH) were determined by the method of radioimmunoassay (RIA). Reverse transcriptase polymerase chain reaction (RT-PCT) technology was used to evaluate the expression of estrogen receptor (ER) in bone. Bone mineral density (BMD) was measured by double energy X-ray absorption (DEXA).

RESULTSIn the climacteric rats, BMD, serum E(2), ER mRNA expression in bone decreased remarkably, and serum FSH, LH and apoptosis of ovarian granulose cells increased obviously. After treating with GNK, all the indexes were reversed except serum E(2). The increase of E(2) was not significant.

CONCLUSIONGNK is effective on climacteric osteoporosis female rats. Its role is performed not by increasing serum E(2) but by enhancing ER in the bone and inhibiting apoptosis of ovarian granulose cells. GNK can deter further exhaustion of ovarian function.

Age Factors ; Animals ; Apoptosis ; Bone Density ; drug effects ; Bone and Bones ; drug effects ; metabolism ; Climacteric ; Drugs, Chinese Herbal ; pharmacology ; Estradiol ; blood ; Female ; Follicle Stimulating Hormone ; blood ; Hormone Replacement Therapy ; Hormones ; blood ; Luteinizing Hormone ; blood ; Osteoporosis ; metabolism ; Ovary ; drug effects ; physiology ; Rats ; Receptors, Estrogen ; biosynthesis

AIMTo investigate the effects of 17beta-estradiol (E2), Peganum harmala extract (PHE) and caloric restriction (CR) on various testis parameters during aging.

METHODSTwelve month-old male rats were treated for 6 months with either E2 or PHE, or submitted to CR (40%).

RESULTSOur results show that estrogens and CR are able to protect the male gonad by preventing the decrease of testosterone and E2 levels as well as the decrease of aromatase and estrogen receptor gene expressions. Indeed, E2, PHE and CR treatments induced an increase in the superoxide dismutase activities and decreased the activity of testicular enzymes: gamma-glutamyl transferase, alkaline phosphatase, lactate deshydrogenase as well as the aspartate and lactate transaminases in aged animals. In addition, the testicular catalase and gluthatione peroxidase activities were enhanced in E2, PHE and CR-treated rats compared to untreated animals at 18 months of age. Moreover, the positive effects of estradiol, PHE and CR were further supported by a lower level of lipid peroxidation. Recovery of spermatogenesis was recorded in treated rats.

CONCLUSIONBesides a low caloric diet which is beneficial for spermatogenesis, a protective antioxydant role of estrogens is suggested. Estrogens delay testicular cell damage, which leads to functional senescence and, therefore, estrogens are helpful in protecting the reproductive functions from the adverse effects exerted by reactive oxygen species (ROS) produced in large quantities in the aged testis.

Aging ; physiology ; Animals ; Antioxidants ; metabolism ; Aromatase ; biosynthesis ; genetics ; Caloric Restriction ; Estradiol ; metabolism ; pharmacology ; Estrogens ; pharmacology ; Lipid Peroxidation ; drug effects ; Male ; Oxidative Stress ; drug effects ; Peganum ; chemistry ; Plant Extracts ; pharmacology ; RNA ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; Receptors, Estrogen ; biosynthesis ; genetics ; Testis ; drug effects ; enzymology ; growth & development ; Testosterone ; metabolism ; Thiobarbituric Acid Reactive Substances ; metabolism