• 생명과학회지
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• 제30권6호
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• pp.550-554
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• 2020

이 연구의 목적은 시험관내 TCD (테스토스테론 화합물 검출) 시스템으로 8개 한약재의 테스토스테론 유사 활성을 검색하는 것이었다. 녹각상, 백수오, 야관문, 홍삼, 토령, 민들레, 남가세, 호로파의 열수 추출물과 80% 에탄올 추출물을 5, 50, 500 ㎍/ml의 농도로 in vitro TCD 시스템에 적용하였다. 80% 에탄올 추출물의 테스토스테론 유사 활성은 일반적으로 농도에 비례하였고, 음성대조군보다 0.9 내지 5.0 배 더 높았다. 호로파와 야관문은 동일한 농도에서 우수한 테스토스테론 유사 활성을 보였으며 민들레와 홍삼이 그 뒤를 이었다. 토령과 남가세는 낮은 테스토스테론 유사 활성을 보였으며 녹각상과 백수오는 매우 낮은 활성을 나타냈다. 양성대조군 5α-DHT의 활성과 비교하면, 호로파 및 야관문의 80% 에탄올 추출물은 500 ㎍/ml에서 10^-8 M 5α-DHT보다 높은 테스토스테론 유사 활성을 나타냈다. 열수 추출물은 80% 에탄올 추출물보다 낮은 테스토스테론 유사 활성을 보였고, 농도에 따른 활성의 증가도 에탄올 추출물보다 낮았다. 호로파, 야관문 및 민들레 등의 80% 에탄올 추출물이 테스토스테론 관련 유전자의 전사 효율을 향상시킬 수 있음이 확인되었다. 따라서, 본 연구는 남성 갱년기 증상의 개선에 필요한 건강 기능성 식품 재료 또는 의약 재료를 개발하는 데 참고가 될 수 있을 것이다.

• 환경위생공학
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• 제21권2호
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• pp.30-35
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• 2006

The purpose of this study finds out the effect of red ginseng extract (1.0g/kg) on TBTO (10, 20, and 40mg/kg) which poisons against some organs like thyroids gland, liver, kidney, testis, ovary, sex hormone activity of rats are examined by gastric tubing for 3 weeks. The weight of each organ in treated group were increased, especially liver in female and those of testis in males were significantly increased at 10, 20 and 40mg/kg (P<0.05, P<0.01). In case of sex hormone activity of each sex, the estradiol activity of female and testosterone activity of males were significantly decreased rather than the control group (P<0.05, P<0.01) According to between the TBTO treated group and 10+ rGe group of the testosterone activity each sex were significantly increased (P<0.01).

• 생명과학회지
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• 제29권10호
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• pp.1159-1163
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• 2019

남성호르몬 감소와 관련된 남성갱년기에 대한 관심이 고조되고 있지만, 남성호르몬의 정량을 위해 항체를 이용하는 고가의 kit가 이용되고 있다. 본 연구에서는 in vitro 전사 활성 시험법을 이용하여 남성 스테로이드호르몬의 활성 혹은 농도를 검증하는 시스템을 구축하였다. 테스토스테론-AR (androgen receptor) 복합체와 반응하는 ARE-AdE1bTATA 염기서열이 삽입되고 리포터로 luciferase를 발현하는 테스토스테론 유사활성 검증 리포터 플라스미드인 pGL2-Neo-ARE-AdE1BTATA를 제조하고, 인체 전립선암 세포인 LNcap-LN3 세포에 stable transfection을 실시하였다. 구축된 LNcap-LN3/pGL2-Neo-ARE-AdE1BTATA TCD (testosterone compound detection) 시스템은 표준물질인 테스토스테론의 $10^{-13}{\sim}10^{-8}M$ 범위에서 농도 증가에 비례하는 정량성을 보였다. 이 연구에서 확립된 in vitro TCD 시스템을 이용하면 천연물 유래 테스토스테론 유사물질 및 테스토스테론 저하물질의 대량 탐색 등이 가능할 것이므로, 건강기능성 식품이나 의약품 신소재의 개발에 기여할 것이다.

• 한국환경보건학회지
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• 제25권3호
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• pp.89-93
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• 1999

The purpose of this study finds out the effect of red ginseng extract(1.0 g/kg) on bis(tributyltin)oxide(TBTO)(10, 20 and 40 mg/kg) which poisons against some organs like thyroid gland, liver, kidney, testis, ovary. Serum immunity and sex hormone activity of rats are examined by gastric tubing for 3 weeks. The weights of each in treated group were increased, especially liver in females and those of testis in males were signignificantly increased at 10, 20 and 40 mg/kg (P<0.05, P<0.01). In group treated with TBTO(10, 20 and 40 mg/kg) for females, the serum IgG level was significantly reduced in comparison with control group(P<0.05). The group of females which were poisonned by TBTO and rGe was significantly recovered in TBTO 10 mg/kg + rGe 1.0 g/kg and TBTO 20 mg/kg + rGe 1.0 g/kg. In case of sex hormone activity of each sex, the estradiol activity of females and testosterone activity of males were signignificantly decreased rather than the control group. And control group by bis(tributyltin) oxide and red ginseng extract is just increased estiradiol activity.

• Advances in Traditional Medicine
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• 제6권2호
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• pp.86-95
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• 2006

The ethanol extract of the Crotalaria juncea seeds, which showed promising antispermatogenic and antiandrogenic activities in albino mice, was taken up further for the isolation of the active fractions present in it. Two fractions that were obtained from thin layer chromatography were subjected for testing to know their antispermatogenic and antiandrogenic activities. After preliminary trials the fraction I showed maximum antifertility activity at the dose level of 200 mg/kg body weight when administered orally to the rats for 50 days. The fraction I was found to affect spermatogenesis as well as the endocrine functions of the testis as indicated by gravimetric, histopathological and biochemical changes. Further this fraction has caused degenerative changes in the seminiferous tubules and Leydig cells of the testis. The accessory reproductive organs like epididymis, seminal vesicles, vas deferens, prostrate, Cowper's gland and Levator Ani muscle showed significant malfunction. Cauda epididymal sperm count and sperm motility were reduced significantly. The treatment has also resulted in increase in the cholesterol level and alkaline phosphatase activity, and decrease in protein, glycogen, sialic acid contents and acid phosphatase activity in testis. It is noteworthy that RIA studies have shown significant reduction in serum FSH, LH and testosterone. Scanning electron microscopic observations revealed abnormalities in sperm structure.

• Journal of Life Science
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• 제11권2호
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• pp.103-107
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• 2001

The equine chorionic gonadotropin (eCG) subunits $\alpha$ and ${\beta}$ are transcribed from different genes and associate noncovalently to form the bioactive eCG heterodimer. Dimerization is rate limiting for eCG secretion, and dissociation leads to hormone inactivation. The correct conformation of the heterodimer is alto important for efficient secretion, hormone-specific post-translational modifications, receptor binding and signal transduction. To determine whether ${\alpha}$ and ${\beta}$ subunits can be synthesized as a single polypeptide chain (tethered-eCG) and also display biological activity, the tethered-eCG molecule by fusing the carboxyl terminus of the eCG ${\beta}$-subunit to the amino terminus of the af-subunit was construe-ted and transfected into chinese hamster ovary (CHO-Kl) cells. LH- and FSH-like activities were assayed in terms of testosterone production and aromatase activity in primary cultured rat Leydig cells and granulosa cells, respectively. The tethered-eCG was efficiently secreted and showed similar LH-like activity to the dimeric eCG ${\alpha}$/${\beta}$ and native eCG. FSH-like activity of the tethered-eCG was also shown similarly in comparison with the native and wild type eCG ${\alpha}$/${\beta}$. Our data for the first time suggest that the tethered-eCG can be expressed efficiently and the produced product by the CHO-K1 cells is fully LH- and FSH-like activities in rat in vitro bioassay system. Our results also suggest that this molecular can imply particular models of FSH-like activity not LH-like activity in the eCG. Taken together, these data indicate that the constructs of tethered molecule will be useful in the study of mutants that affect subunit association and/or secretion.

• 환경위생공학
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• 제16권3호
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• pp.8-13
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• 2001

This study was carried out effect of red ginseng extract(1.0g/kg) on organotin compounds (10, 20 and 40 mg/kg) which poisons against some organs like thyroid gland, liver, kindey, testis, ovary, serum immuinty and sex hormone activity of rats were examinned by gastric tubing for 3 weeks. The weight of each organ in TBTO treated group were significantly increased other organs which excepted kinedy in males and only liver in females.(p<0.05, p<0.01). In case of Immunity activity of each sex, IgM level was small change comparsion with that of control group in all sex. but IgG level was significantly decreased females rather than males comparsion with that of control group.(p<0.05, p<0.01) In case of sex hormone activity, the testosterone activity of males and the estradiol activity of females were significantly decreased rather than the control group. on the other hand, red singsong treated group was only significantly increased estradiol activity.( p<0.05, p<0.01)

• Toxicological Research
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• 제18권1호
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• pp.59-64
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• 2002

Substantial evidences have been accumulated about the hormone-like effects of exogenous substances such as pesticides and industrial chemicals during past years. The effects of these substances on the endocrine system are believed to be either enhancing or reducing of various endocrine action. It is necessary to identify putative causal agents by the batter system and to assess their ability to disrupt the endocrine system. A variety of in vitro and In vivo approaches have been used to determine the androgenic effects of environmental chemicals. To establish the method for assessment of the putative endocrine disruptors with androgenic activity, we carried out the cell proliferation assay by MTS method after treatment with the various concentration of testosterone in LNCaP cells (human prostatic cancer cell line) and also observed the expression of androgen-related genes by quantitative RT-PCR. In the cell proliferation assay, the results showed that the grouth of LNCaP cells increased within level of at least 10pM testosterone. We measured by quantitative RT-PCR method on the effects of testosterone on mRNA expression of androgen receptor (AR), prostate-specific antigen (PSA), bone morphogenetic protein (BMP) and BMP receptor (BMPR) In LNCaP cells. The results demonstrated that mRNA expression of PSA and BMPR-IB was observed differently within level of at least 0.01 pM testosterone compared with non-treated control. These observations suggest that the detection of PSA and BMPR-IB mRNA by the quantitative RT-PCR in LNCaP cells is very sensitive method to identify the endocrine disruptors to have the androgenic effects.

• Environmental Analysis Health and Toxicology
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• 제31권
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• pp.10.1-10.8
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• 2016

Objectives Aromatase inhibitors that block estrogen synthesis are a proven first-line hormonal therapy for postmenopausal breast cancer. Although it is known that standardized extract of Ginkgo biloba (EGb761) induces anti-carcinogenic effects like the aromatase inhibitors, the effects of EGb761 on steroidogenesis have not been studied yet. Therefore, the effects of EGb761 on steroidogenesis and aromatase activity was studied using a H295R cell model, which was a good in vitro model to predict effects on human adrenal steroidogenesis. Methods Cortisol, aldosterone, testosterone, and $17{\beta}$-estradiol were evaluated in the H295R cells by competitive enzyme-linked immunospecific assay after exposure to EGb761. Real-time polymerase chain reaction were performed to evaluate effects on critical genes in steroid hormone production, specifically cytochrome P450 (CYP11/ 17/19/21) and the hydroxysteroid dehydrogenases ($3{\beta}$-HSD2 and $17{\beta}$-HSD1/4). Finally, aromatase activities were measured with a tritiated water-release assay and by western blotting analysis. Results H295R cells exposed to EGb761 (10 and $100{\mu}g/mL$) showed a significant decrease in $17{\beta}$-estradiol and testosterone, but no change in aldosterone or cortisol. Genes (CYP19 and $17{\beta}$-HSD1) related to the estrogen steroidogenesis were significantly decreased by EGb761. EGb761 treatment of H295R cells resulted in a significant decrease of aromatase activity as measured by the direct and indirect assays. The coding sequence/Exon PII of CYP19 gene transcript and protein level of CYP19 were significantly decreased by EGb761. Conclusions These results suggest that EGb761 could regulate steroidogenesis-related genes such as CYP19 and $17{\beta}$-HSD1, and lead to a decrease in $17{\beta}$-estradiol and testosterone. The present study provides good information on potential therapeutic effects of EGb761 on estrogen dependent breast cancer.

• 농약과학회지
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• 제12권3호
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• pp.207-214
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• 2008

내분비장애 추정물질의 분류를 위해 많은 시험법이 연구되고 있는데 추후 내분비장애 추정물질로 분류된 기등록 농약에 대한 자료요구 또는 신규 등록농약에 대한 등록기준의 추가 등을 고려하여 OECD와 EPA에서 권장하는 시험법을 확립하고자 본 연구를 수행하였다. 시험약제를 30일간 경구 투여하여 조사한 결과, alachlor 투여 수컷에서 25 mg/kg/day, 50 mg/kg/day에서 고환과 갑성선의 중량이 증가하였다. Alachlor 투여 암컷에서는 질의 중량이 25, 50 mg/kg/day에서 감소하였고 25 mg/kg/day에서 갑상선의 중량이 감소하였다. Alachlor 투여 암컷 25, 50 mg/kg/day에서 주요 갑상선 호르몬인 T4와 성호르몬 testosterone이 감소하였다. 따라서 pubertal assay 결과 alachlor는 갑상선 호르몬성 영향이 의심되었다. 시험세포를 이용한 시험 결과, 시험약제를 1 nM에서 1000 nM까지 처리하였을 때 음성대조군과 비교하여 alachlor는 100-134%의 갑상선 호르몬성 영향을 나타내었다. 따라서 세포를 이용한 시험에서는 alachlor에 의한 갑상선 호르몬성 영향이 나타나는 것으로 판단되었다. 항갑상선 호르몬성 영향 시험에서는 시험약제 100 nM과 T4의 혼합 처리시 alachlor는 항갑상선 호르몬성 영향은 나타나지 않았다.