• Journal of Life Science
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• v.23 no.9
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• pp.1155-1162
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• 2013

Estrogen like activities were evaluated using ethanol and hot water extracts of gongjindan (GJD) and 3 types of brown algae by an in vitro detection system. Laminaria japonica and Undaria pinnatifida represented statistically significant (p<0.001) estrogen like activities, while GJD and Ecklonia stolonifera did not. Furthermore, activities of hot water extracts ($500{\mu}g/ml$) of Laminaria japonica and three mixture (Ecklonia stolonifera : Undaria pinnatifida : Laminaria japonica, 1: 1: 1) were stronger than that of $10^{-7}M$ $17{\beta}$-estradiol. These results suggest that Laminaria japonica contain estrogen like compounds. Based on these results, the prepared seaweed GJD were used for animal experiments about the effects on inhibition of platelet aggregation and serum lipid levels in ovariectomized rats. 9-week Old female SD-rats were randomly assigned to 3 groups as sham-operated rats, ovariectomized rats and ovariectomized rats that were treated with GJD. The rats were placed on prescribed diets for 5 weeks following ovariectomy. Total cholesterol and triglyceride contents on serum decreased in the SHAM group compared to the OVX-CON group. 5 weeks feeding of GJD resulted in significant lowering of triglyceride and a decreasing tendency of total cholesterol level. The level of HDL-cholesterol on serum increased significantly by feeding diets containing the GJD. The ability of platelet aggregation of group treated with GJD was less than that of the OVX-CON group. These results suggest that the beneficial effects of seaweed gongjindan may be used to improve menopausal syndrome.

• Journal of Applied Biological Chemistry
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• v.59 no.4
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• pp.305-311
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• 2016

This study investigated the effects of a mixture of fenugreek seeds and Lespedeza cuneata extracts on testosterone synthesis in TM3 cells that were oxidatively stressed with $H_2O_2$. In order to oxidatively stress TM3 cells, the cells were treated with $50{\mu}M$ hydrogen peroxide for 4 hr in serum-free media. Yagwanmun-horopa mixture (YHM) showed neither cytotoxicity nor increment of cell proliferation in the oxidatively stressed TM3 cells in any concentration. When the cells were treated with hydrogen peroxide, testosterone levels decreased, but the testosterone level was returned to that of the control level in the presence of YHM. In order to find out the reasons for the increase of testosterone, the expression of the genes involved in the synthesis or disintegration of testosterone. On the other hand, the levels of $3{\beta}$-HSD4 and 17, 20-desmorase, which are involved in testosterone synthesis, were decreased through the use of hydrogen peroxide and were recovered through YHM treatment. Aromatase and $5{\alpha}$-reductase2, which convert testosterone to estradiol and dihydrotestosterone, respectively, were increased through the use of hydrogen peroxide, and were returned to control level through YHM treatment. These results suggest that YHM does not affect TM3 cell proliferation. However, YHM increases the expression of testosterone-synthesizing enzyme, which was decreased through oxidative stress, and decreases the expression of testosterone- converting enzyme, which was increased through oxidative stress. Therefore, it is reasonable that YHM has strong recovery activity on testosterone to normal level, even in the oxidatively stressed TM3 cells which mimics the andropause state.

• Environmental Analysis Health and Toxicology
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• v.20 no.3 s.50
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• pp.195-203
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• 2005

Cytochrome P45O 17$\alpha$-hydroxylase (CYPI 7) and cytorhrome P45O aromata.ie (CYPI 9) are key steroidogenic enzymes in androgen and estrogen synthesis. ThiL study evaluated the effects of nonylphenol (NP) on CYP17 and CYP19 expression in the ovary of Sprague-Dawley rats. All female rats were administered orally with the vehicle (control, corn oil), diethylstilbestrol (DES, 5.0 $\mu$g/kg) and NP (50, 100, or 200 mg/kg/day), which was startinB when they were weaned at 21 days of age for 20 days. Twenty four hours after final dose, the animals were anelthetized with ether. Significant decreases in the uterus (wet weight) were observed with 5.0 $\mu$g/kg/day DES (78$\%$, of control) and 200 mg/kg/day NP (62$\%$ of control), respectively Additionally, ovarian weight was significantly decreased with 5.0 $\mu$g/kg/day DES (63$\%$ of control) and 200 mg/kg/day NP (72$\%$ of control). The serum estradiol levels were sligHtly lower in DES and high dose NP treatment groups, but the 74 levels were not affected by DES and NP. The expression of the ovarian CYP19 gene increased with low doses (50 and 100 mg/kg/day) of NP. while DES and high dose oi NP (200 mg/kg/day) did not affect on the CYP19 mRNA levels. In contrast to the CYP19 gene, the CYP17 gene expreLsion level was significantly down-regulated by the DES and 200 mg/ks/day NP. This result suggestE that NP inhibits ovarian estrogen synthelis by supprelsing CYP17 mRNA efprelsion, And different mechanisml might exist for the expression of Lteroidogenic CYP17 and CYP19 genes in the ovary of Sprague-Dawley rats in response to NP.

• Journal of Life Science
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• v.23 no.12
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• pp.1501-1508
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• 2013

This study investigated antioxidant activity and the lipid content of serum for the possible outcome of improving the activity of Ishige okamurae extracts in ovariectomized rats. The antioxidant effects of an Ishige okamurae water extract and an Ishige okamurae ethanol extract were measured by evaluating DPPH free radical scavenging activity and SOD-like activity. Fifty, seven-week old female Sprague Dawley rats were randomly assigned to five groups as follows: sham-operated rats (SHAM), ovariectomized rats (OVX-CON), ovariectomized rats that were treated with 17-beta-estradiol (200 ${\mu}g/kg/day$), and ovariectomized rats that were treated with Ishige okamurae extracts (50 mg/kg/day and 200 mg/kg/day, respectively). The diets were fed to the rats for seven weeks after ovariectomy. The antioxidant activities of the water and ethanol extracts of Ishige okamurae increased in a dose-dependent manner, and the ethanol extract was found to be higher than the water extract. Therefore, we examined the effect of an Ishige okamurae ethanol extract on total serum cholesterol, triglycerides, HDL-cholesterol, and LDL-cholesterol levels, and anti-platelet aggregation. The total-cholesterol and triglyceride content of the serum increased in the OVX-CON group compared to the SHAM group, but supplementation with the Ishige okamurae ethanol extract caused these factors to decrease. Notably, the serum LDL-cholesterol concentration in the supplemented 200 mg/kg/day Ishige okamurae ethanol extract group was significantly more reduced than it was in the OVX-CON group. In addition, the platelet aggregation ability was lower in the groups treated with Ishige okamurae than it was in the OVX-CON group. According to these results, the effects of Ishige okamurae extract on serum lipid content in ovariectomized rats were illuminated.

• Development and Reproduction
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• v.5 no.2
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• pp.175-180
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• 2001

The ectopic expression of gonadotropin releasing hormone(GnRH and luteinizing hormone(LH) in several tissues is a quite intriguing phenomenon. Recently, the presence of GnRH and its receptor has been clearly demonstrated in rodents and human mammary gland. In this context, one can postulate that the presence of local circuit composed of GnRH and LH in the gland. The present study was undertaken to elucidate whether there is a correlation between the LH expression in rat mammary gland and physiological status during the process of mammary differentiation. LH contents in mammary gland from cycling to weaning rats were measured by radioimmunoassay(RIA). In cycling rats, changes of the LH level in both serum and mammary gland showed similar pattern as the highest level in proestrus and the lowest level in diestrus II stage. While the serum LH levels were fluctuated from pregnant through involution stage, a sharp decline of mammary LH contents was observed in the lactating rats. This decrement was recovered in involuting rats to the level of proestrus stage. Reverse transcription-polymerase chain reaction (RT-PCR) and Southern blot analyses demonstrated that the transcriptional activities of the mammary LH and GnRH were increased from diestrus I stage to estrus stage, and the increased levels were maintained in pregnant, lactation and involution stages. To test the hypothesis that the alteration in mammary LH expression might be steroid-dependant, ovariectomy(OVX) and steroid supplement model was employed. As expected, supplement of estradiol(E$_2$) after OVX remarkably decreased serum LH level compared to that in serum from vehicle-only treated rats. Likewise, administration of E$_2$ significantly reduced the mammary LH content. The present study demonstrated that (i) the LH expression in mammary gland could be altered by some physiological parameters such as estrous cycle, pregnancy, lactation and involution, and (ii) ovarian steroid especially estrogen seems to be one of major endocrine factors which are responsible for regulation of mammary LH expression.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.4
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• pp.463-470
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• 2011

Gonadotropin-releasing hormone receptor (GnRHR) gene is expressed at the anterior pituitary gland and plays a key role in gonad development. This study aimed to investigate molecular genetic characteristics of the GnRHR gene and elucidate the effects of single nucleotide polymorphisms (SNPs) of GnRHR gene on sex steroid level in Japanese flounder (Paralichthys olivaceus). We used polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) and sequencing of the GnRHR gene in 75 individuals. We identified three SNPs in the GnRHR gene: P1 locus (C759A and C830T) in the coding region of exon2 which were both linked together and P2 locus (G984T) in the coding region of exon3, which added a new transcript factor (ADR1) and a new methylation site (CG). Only C830T of P1 leads to amino acid changes Thr266Ile. Statistical analysis showed that P1 was significantly associated with $17{\beta}$-estradiol ($E_2$) level (p<0.01) and gonadosomatic index (GSI) (p<0.05). Individuals with genotype BB of P1 had significantly higher serum $E_2$ levels (p<0.01) and GSI (p<0.05) than those of genotype AA or AB. Another SNP, P2, synonymous mutation, was significantly associated with GSI (p<0.05). Individuals with genotype AB of P2 had significantly higher GSI (p<0.05) than that of genotype AA. In addition, there was a significant association between one diplotype based on three SNPs and reproductive traits. The genetic effects for both serum $E_2$ level and GSI of diplotype D4 were super diplotypes (p<0.05). These results suggest that the SNPs in Japanese Flounder GnRHR are associated with $E_2$ level and GSI.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.4
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• pp.302-306
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• 2000

Gonad and blood samples were taken from cultured greenling (Hexagrammos otakii), and changes of steroid hormone levels in serum were examined in relation to the gonadosomatic index (GSI). GSI of female and male were not changed significantly between March and August and then began to increase from September showing the highest value in January. Hepatosomatic index (HSI) of female was the highest in December,'then decreased gradually until May. $Estradiol-l7{\beta} (E_2)$ level in blood of female reached to the highest level in December, then decreased rapidly in January. The level was maintained low until August and started to increase In September. Testosterone (T) level in blood of female was similar to trend of $E_2$ in female. $17{\alpha}2{\beta}-dihydroxy-4-pregnen-3-one (17{\alpha}20{\brta}-OHP)$ level in blood of female reached to the maximum in January, and then decreased rapidly from Febuary and maintained low until July. T level in blood of male reached to the highest value in December, and it started to decrease from January and maintained low until July, after then increased again, while $17{\alpha}20{\beta}-OHP$ level of male was similar to trend of female's.

• Journal of Ginseng Research
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• v.44 no.6
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• pp.790-798
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• 2020

Background: Beneficial effects of Korean Red Ginseng (KRG) on polycystic ovarian syndrome (PCOS) remains unclear. Methods: We examined whether pretreatment (daily from 2 hours before PCOS induction) with KRG extract in water (KRGE; 75 and 150 mg/kg/day, p.o.) could exert a favorable effect in a dehydroepian-drosterone (DHEA)-induced PCOS rat model. Results: Pretreatment with KRGE significantly inhibited the elevation of body and ovary weights, the increase in number and size of ovarian cysts, and the elevation of serum testosterone and estradiol levels induced by DHEA. Pretreatment with KRGE also inhibited macrophage infiltration and enhanced mRNA expression levels of chemokines [interleukin (IL)-8, monocyte chemoattractant protein-1), proinflammatory cytokines (IL-1β, IL-6), and inducible nitric oxide synthase in ovaries induced by DHEA. It also prevented the reduction in mRNA expression of growth factors (epidermal growth factor, transforming growth factor-beta (EGF, TGF-β)) related to inhibition of the nuclear factor kappa-light-chain-enhancer of activated B cell pathway and stimulation of the nuclear factor erythroid-derived 2-related factor 2 pathway. Interestingly, KRGE or representative ginsenosides (Rb1, Rg1, and Rg3(s)) inhibited the activity of inflammatory enzymes cyclooxygenase-2 and iNOS, cytosolic p-IκB, and nuclear p-nuclear factor kappa-light-chain-enhancer of activated B in lipopolysaccharide-induced RAW264.7 cells, whereas they increased nuclear factor erythroid-derived 2-related factor 2 nuclear translocation. Conclusion: These results provide that KRGE could prevent DHEA-induced PCOS via antiinflammatory and antioxidant activities. Thus, KRGE may be used in preventive and therapeutic strategies for PCOS-like symptoms.

• Animal cells and systems
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• v.16 no.2
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• pp.127-134
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• 2012

We investigated the effects of fadrozol, an aromatase inhibitor (AI), and $17{\alpha}$-methyltestosterone (MT) on the induction of sex change in juvenile longtooth grouper $Epinephelus$ $bruneus$, via histological observation of gonads. Changes in the mRNA expression of GtH subunits (FSH-${\beta}$ and LH-${\beta}$) in the pituitary, and estradiol-$17{\beta}$ (E2) and 11-ketotestosterone (11-KT) levels in the blood were also surveyed after AI and MT treatment. Juvenile longtooth groupers ($113{\pm}17g\;body\;weight$; $16.2{\pm}1.2cm\;body\;length$) received intramuscular injections of AI at 3 (3-AI) and 5 (5-AI) mg/kg BWdoses and MT at a 5 mg/kg BW (5-MT) dose. At week 7 post-injection, 3-AI and 5-MT oocytes were degenerated, and gonads of the 5-AI group initiated spermatogenesis. At week 21 post-injection, 3-AI- and 5-MT-treated gonads contained spermatogonia and spermatocytes, while 5-AI treatment induced advanced stages of spermatogenesis. The serum E2 level showed no significant differences throughout the experimental period, whereas that of 11-KT was significantly elevated in the 5-AI group at weeks 7 and 21 post-injection. A significant increase in the expression of FSH-${\beta}$ mRNA was evident in the 5-AI group at week 21 post-injection. In contrast, LH-${\beta}$ mRNA expression did not significantly differ among groups during the experimental period. These results imply that sex change has two stages in the longtooth grouper. In the first stage, oocytes are degenerated by the stimulation by 11-KT, and in the second stage spermatogenesis occurs, owing to the co-effects of 11-KT and FSH-${\beta}$.

• Korean Journal of Veterinary Research
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• v.32 no.1
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• pp.15-24
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• 1992

The effects of CC the ovulatory response, oocyte normality, ovarian steroidogenesis and subsequent embryo developmental potential were examined in PMSG-treated rats. On Days of 25~27 of age, immature female Sprague Dawley rats were treated with three different doses(0.05, 0.1 or 1.0mg /day) of clomiphene citrate or vehicle. The females subsequently received 4IU PMSG on Day 28 and/or 10IU hCG on Day 30, and were killed on Day 31. Some females given 0.1mg CC or vehicle with 4IU PMSG were then mated and killed on Days 2, 3, 4 and 5 of pregnancy. Compared to vehicle(control) group, by increasing the doses of CC, there were a significant decrease in the ovulatory response as judged by both the proportion of rats ovulating and the mean number of oocytes per rat and a marked reduction of ovarian weight. The increasing doses of CC substantially promoted the degeneration(%) of oocytes ovulating in a dose-dependent manner. The CC-mediated inhibitions of the ovulatory response and ovarian weight were oompletely overcome by a subsequent treatment of hCG. Increasing doses of CC resulted in a siginificant elevation of serum estradiol with the decreased levels of progesterone and androgens. The additive treatment with hCG was effective to reduce the elevation of estradiol and to increase the reduction of progesterone produced by high dose(1.0mg) of CC. The preimplantation embryos recovered from 0.1mg CC-treated pregnant rats demonstrated a progressive early loss from Day 3 of pregnancy with a significant increase in the percentage of degeneration during all periods examined, compared to controls. The rate of progressive embryo cleavage in the CC-treated rats were slower than that in controls from Day 3 of pregnancy. Additionally, the percentage of the cleaved embryos recovered from the CC-treated rats remained significantly lower consistently from Day 2 of pregnancy, compared to control regimen. These results demonstrate a possible mechanism of CC-mediated inhibition of ovulatory response in the rats which may include the attenuation or blockade of the endogenous secretion of gonadotropins and also suggest that its detrimental effects observed on oocyte normality and embryonic development may be caused by abnormal follicular steroidogenesis( especially elevated estradiol) preceding fertilization.