• Clinical and Experimental Reproductive Medicine
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• v.11 no.2
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• pp.17-25
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• 1984

Hypothalamic-pituitary function in patients of 6 selected groups of amenorrhea was evaluated by performing premarin test. Selected amenorrheic patients were divided into 6 groups of Turner's syndrome(5), adrenogenital syndrome(3), Sheehan's syndrome(4), moderate hyperprolactinemia(3), severe hyperprolactinemia(9) and functional oligoamenorrhea(9) the diagnoses of which were performed according to modified our own protocol for management of amenorrheic patients. As control 20 normally cycling women in mid follicular phase determined by their symptothermal charts during last 6 months designed by WHO were compared. The premarin test which is one of the tests evaluating the hypothalamic-pituitary function by the principle of negative and positive feed back effect's of estrogen was performed by injecting 20 mg of premarin in volus intravenously. The levels of serum LH before, 24, 48, 72, 96 and 120 hours after injection of premarin were measured by double antibody technique radioimmunoassay the reagents of which were supplied by WHO. The results were as follows: 1. Both negative and positive feed back effects by exogenous estrogen were well preserved even in the patients of gonadal dysgenesis although the baseline levels were much higher than normal. 2. In the patients of Sheehan's syndrome one could observe the minimal response of feed back effect in the case with minimal pituitary function. 3, Androgens in adrenogenital syndrome and prolactin in hyperprolactinemia may suppress mainly the positive feed back effect rather than the negative one. The suppressive effect can be abolished by proper treatments which can eliminate those suppressive hormones. 4. This premarin test may be beneficial for predicting the result of clomiphene in ovulation induction.

• Development and Reproduction
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• v.13 no.3
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• pp.133-143
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• 2009

Pivotal roles of steroid hormones in uterine endometrial function are well established from the mouse models carrying the null mutation of their receptors. Literally androgen belongs to male but interestingly it also detected in female. The fluctuations of androgen levels are observed during reproductive cycle and pregnancy, and the functional androgen receptor is expressed in reproductive organs including uterus. Using high throughput methodology, the downstream genes of androgen have been isolated and revealed correlations between other steroid hormones. In androgen-deficient mice, uterine responses to exogenous gonadotropins are impaired and the number of pups per litter is reduced dramatically. As expected androgen has important role in decidual differentiation through AR. It regulates specific gene network during those cellular responses. Recently we examined the effects of steroid hormonal complex containing high level of androgen. Interestingly, on the contrary to the androgen-alone administration, the hormonal complex did not disturb the decidual reaction and the pubs did not show any morphological abnormality. It is suspected that the complexity of communication between other steroid hormone and their receptors are the reasons. In summary, androgen exists in female blood and it suggests the importance of androgen in female reproduction. However, the complex interactions with other hormones are not fully understood compared with estrogen and progesterone. The further studies to evaluate the possible role of androgen are needed and important to provide the in vivo rational for the prevention of associated pregnancy complications and help human's health.

• Development and Reproduction
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• v.19 no.1
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• pp.43-51
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• 2015

Connexin (Cx) is a complex which allows direct communication between neighboring cells via exchange of signaling molecules and eventually leads to functional harmony of cells in a tissue. The initial segment (IS) is an excurrent duct of male reproductive tract and expression of numerous genes in the IS are controlled by androgens and estrogens. The effects of these steroid hormones on gene expression in the IS during postnatal development have not extensively examined. The present research investigated expressional modulation of Cx isoforms in the IS by exogenous exposure to estrogen agonist, estradiol benzoate (EB), or androgen antagonist, flutamide (Flu), at weaning age. Two different doses of EB or Flu were subcutaneously administrated in 21-day old of male rats, and expressional changes of Cx isoforms in the adult IS were analyzed by quantitative real-time PCR. Treatment of a low-dose EB ($0.015{\mu}g/kg$ body weight) resulted in an increased expression of Cx31 gene and a decreased expression of Cx37 gene. A high-dose EB ($1.5{\mu}g/kg$ body weight) treatment caused an increase of Cx31 gene expression. Increased levels of Cx30.3 and Cx40 transcripts were observed with a low-dose Flu ($500{\mu}g/kg$ body weight) treatment. Treatment of high-dose Flu (50 mg/kg body weight) led to expressional increases of Cx30.3, 40, and 43 genes. Our previous and present findings suggest differential responsiveness on gene expression of Cx isoforms in the IS by androgens and estrogens at different postnatal ages.

• Journal of Veterinary Clinics
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• v.16 no.1
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• pp.42-49
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• 1999

The reproductive effect of $Telazol^{\circledR}$ was studied in pregnant rats. Two different doses (20 or 40 mg/kg/day) were injected on days one of 3, 5 or 8 of pregnancy and 3 consecutive days of 3~5 or 5~7 in pregnant rats. On days 8 or 21, the rats were anesthetized, the cesarean section was performed, the uterus was carefully examined, and the numbers of corpora lutea, implantation sites and viable fetuses, fetal and placental weights and pre-and post-implantation losses in rats were surveyed. Simultaneously the concentrations of sodium and potassium of the plasma, uterine and amniotic fluids were determined. And gestation period in pregnant rats, postnatal offspring survival and body weight gains were evaluated. Administration of $Telazol^{\circledR}$ did not exert a bad influence upon body weight gains in pregnant rats. Reproductive indices were normal. The concentrations of sodium and potassium of the plasma, uterine and amniotic fluids in pregnant rats remained unchanged. Gestation period of rats, postnatal offspring survival and body weight gains were normal. It is concluded that $Telazol^{\circledR}$ is devoided of significant adverse reproductive effects in pregnant rats.

• Proceedings of the Korean Society of Applied Pharmacology
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• 2003.11a
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• pp.88-88
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• 2003

Cytochrome P-450 3A4 (CYP3A4) is major enzyme in human liver, the role of this is detoxification and metabolizing more than 50% clinical drugs in use. Expression of CYP3A4 is transciptionally regulated by the Pregnenolone X receptor (PXR), of which human form is Steroid and Xenobiotics receptor (SXR). SXR is activated by wide range of endogenous and exogenous compounds, and then induces CYP3A4 gene expression. In the previous study, it has been known that proximal promoter (-864 to +64) does not response to chemical inducers such as pregnenolone 16a-carbonitrile (PCN), Rifampicin, Estrogen in terms of transcription of CYP 3A4 in cultured cells. Here, we developed luciferase reporter gene assay system to detect SXR-based CYP 3A4 transcriptional activity. We have used CYP3A4-Luc plasmid that contains proximal promoter of human CYP3A4 gene upstream of the luciferase gene. We did transient transfection of 3A4-luciferase gene and SXR. In the HepG2 cells transfected with CYP3A4-Luc, when rifampicin treatment was combined with histone deacetylase inhibitor (HDAC Inhibitor), such as Trichostatin A, Hc-toxin and IN 2001 of the luciferase activity was induced 10-20 fold over control.

• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 2003.10a
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• pp.178-178
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• 2003

Cytochrome P-450 3A4 (CYP3A4) is major enzyme in human liver, the role of this Is detoxification and metabolizing more than 50% clinical drugs in use. Expression of CYP3A4 is transciptionally regulated by the Pregnenolone X receptor (PXR), of which human form is Steroid and Xenobiotics receptor (SXR). SXR is activated by wide range of endogenous and exogenous compounds, and then induces CYP3A4 gene expression. In the previous study, it has been known that proximal promoter (-864 to ＋64) does not response to chemical inducers such as pregnenolone 16a-carbonitrile (PCN), Rifampicin, Estrogen in terms of transcription of CYP 3A4 in cultured cells. Here, we developed luciferase reporter gene assay system to detect SXR-based CYP 3A4 transcriptional activity. We have used CYP3A4-Luc plasmid that contains proximal promoter of human CYP3A4 gene upstream of the luciferase gene. We did transient transfection of 3A4-luciferase gene and SXR. In the HepG2 cells transfected with CYP3A4-Luc, when rifampicin treatment was combined with histone deacetylase inhibitor (HDAC Inhibitor), such as Trichostatin A, Hc-toxin and IN 2001 of the luciferase activity was induced 10-20 fold over control.

• Korean Journal of Veterinary Research
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• v.34 no.4
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• pp.745-757
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• 1994

Superovulatory treatment with exogenous gonadotropins adversely affects the uterus through the disruption of the delicate balance of ovarian steroids (estrogens, progestins, androgens). To examine the uterine effects of this treatment, 189 rats were given 4IU, 20IU or 40IU pregnant mare',s serum gonadotropin(PMSG) at 28 days of age and sacrificed every 24h until day 10(D10) post injection. Long term uterine effects were examined in 12 rats treated with 4IU or 40IU PMSG and killed on D30. Adult rat uteri were examined to provide a reference for comparisons. Morphological and histological changes of control (4IU) uteri mimicked those of the adult on a comparable time-course form D2 to D5. Administration of superovulatory doses(20IU, 40IU) of PMSG produced stromal hypertrophy by D2 and focal papillary hyperplasia of the luminal epithelia by D3. Levels of $17{\beta}$-estradiol following 20IU and 40IU PMSG treatment were significantly(p<0.05, p<0.005) elevated above those of controls after D1. Androgen levels of both groups(20IU, 40IU) significantly p<0.05, p<005 increased from baseline on D1 and were maximum between D2 and D3. In the 20IU PMSG group, the hyperplasia gradually regressed after D3 and was absent by D10. The hyperplasia in the 40IU PMSG group, however, had become extensive by D6. It is suspected that preceding elevated levels of estrogen may be responsible for this progressive change. On D 4, the levels of $17{\beta}$-estradiol reached a maximum, which was significantly(p<0.001) greater than both the controls and 20IU PMSG-treated rats. Between D6 and D10, the hyperplasia in 40IU PMSG-treated rats partially regressed. Examination of uteri from D30 revealed no evidence of the hyperplasia. It is suggested that previous exposure to high levels of estrogen and androgens, secondary to superovulation, is possible cause for the observed hyperplasia.

• Archives of Pharmacal Research
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• v.20 no.5
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• pp.465-470
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• 1997

The human cDNA clone UDPGTh2, encoding a liver UDP-glucuronosyltransferase (UDPGT), was isolated from a .gamma.gt 11 cDNA library by hybridization to mouse transferase cDNA clone, UDPGTm1. The two clones had 74% nlicleotide sequence identities in the coding region. UDPGTh2 encoded a 529 amino acid protein with an amino terminus membrane-insertion signal peptide and a carboxyl terminus membrane-spanning region. In order to establish substrate specificity, the clone was inserted into the pSVL vector (pUDPGTh2) and expressed in COS 1 cells. Sixty potential substrates were tested using cells transfected with pUDPGTh2. The order of relative substrate activity was as follows: 4-hydroxyestrone > estriol >2-hydroxyestriol > 4-hydroxyestradiol > $6{\alpha}$-hydroxyestradiol >$5{\alpha}$-androstane-$3{\alpha}$, $11{\beta}$, $17{\beta}$-triol=5${\beta}$-androstane-$3{\alpha}$ ${\beta}$, $17{\beta}$-triol. There were only trace amounts of gulcuronidation of 2-hydroxyestradiol and 2-hydroxyestrone, and in contrast to other cloned transferase, no gulcuronidation of either the primary estrogens and androgens (estrone, $17{\beta}$estradiol/testosterone, androsterone) or any of the exogenous substrates tested was detected. A lineweaver-Burk plot of the effect of 4-hydroxystrone concentration on the velocity of glucuronidation showed an apparent Km of $13{\mu}M$. The unique specificity of this transferase might play an important role in regulating the level and activity of these potent and active estrogen metabolites.

• Molecules and Cells
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• v.25 no.2
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• pp.265-271
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• 2008

Single nucleotide polymorphisms (SNPs) are the most common form of human genetic variation. Non-synonymous SNPs (nsSNPs) change an amino acid. Organic anion transporters (OATs) play an important role in eliminating or reabsorbing endogenous and exogenous organic anionic compounds. Among OATs, hOAT4 mediates high affinity transport of estrone sulfate and dehydroepiandrosterone sulfate. The rapid bone loss that occurs in post-menopausal women is mainly due to a net decrease of estrogen. In the present study we searched for SNPs within the exon regions of hOAT4 in Korean women osteoporosis patients. Fifty healthy subjects and 50 subjects with osteoporosis were screened for genetic polymorphism in the coding region of SLC22A11 (hOAT4) using GC-clamp PCR and denaturing gradient gel electrophoresis (DGGE). We found three SNPs in the hOAT4 gene. Two were in the osteoporosis group (C483A and G832A) and one in the normal group (C847T). One of the SNPs, G832A, is an nsSNP that changes the $278^{th}$ amino acid from glutamic acid to lysine (E278K). Uptake of [$3^H$] estrone sulfate by oocytes injected with the hOAT4 E278K mutant was reduced compared with wild-type hOAT4. Km values for wild type and E278K were $0.7{\mu}M$ and $1.2{\mu}M$, and Vmax values were 1.8 and 0.47 pmol/oocyte/h, respectively. The present study demonstrates that hOAT4 variants can causing inter-individual variation in anionic drug uptake and, therefore, could be used as markers for certain diseases including osteoporosis.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.37 no.3
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• pp.192-196
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• 2004

The in vitro effect of bisphenol A (BPA) on ovarian steroidogenesis of the longchin goby (Chasmichthys dolichognathus) was investigated. Oocytes taken during the maturing phase (vitellogenic, fully vitellogenic or germinal vesicle breakdown stage) were incubated with BPA (100 ng/mL) in the presence of exogenous precursor $^{3}H-17\alpha\;hydroxyprogesterone\;(^{3}H-17\alphaOHP).$ Steroids were extracted from the media and the isolated oocytes, and the extracts were separated and identified by thin layer chromatography and gas chromatography-mass spectrometry. The identities of the major metabolites were progestogens $[17{\alpha}-hydroxy,20{\alpha}-dihydroprogesterone\;(17{\alpha}20{\alpha}OHP)\;and\;17{\alpha}-hydrxy,20{\beta}-dihydroprogesterone\;(17{\alpha}20{\beta}OHP),$ androgens [androstenedione (A4) and testosterone (T)] and estrogens [estrone $(E_1)\;and\;estradiol-17{\beta}(E_2)].$ BPA treatment inhibited production of estrogens in all the maturing phases and progestogens in the germinal vesicle migrating stage. Percentage yield of estrogens was decreased with increased yield of androgens. In conclusion, BPA had an inhibitory effect on the conversion of $^3H-17\alphaOHP$ to estrogens and progestogens. These results demonstrate that BPA can act either estrogenic or anti-estrogenic effects.