• 생명과학회지
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• 제28권6호
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• pp.648-655
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• 2018

신령버섯 액체배양추출물(Agaricus blazei mycelial liquid culture extract: ABMLCE)과 eritadenine (EA)의 mouse 정소 Leydig TM3 세포에서 testosterone (TS) 생성에 관한 연구를 정상 조건과 산화스트레스 조건에서 수행하였다. 정상 조건에서는 TM3 세포를 DMEM 배지에 배양하면서 EA (0~100 ppm)와 EA (0~50 ppm) + ABMLCE (10 ppm)를 24 hr처리하였다. 산화스트레스 조건에서는 TM3 세포를 $H_2O_2$ ($50{\mu}M$)를 4 hr 처리하고 정상 조건과 동일하게 처리하였다. DMEM 배양물에 함유된 TS 함량, HSD3B2 효소 활성, $5{\alpha}-R2$ 효소 활성과 NO 함량을 assay kit를 사용하여 측정하였다. EA는 정상 조건이나 산화스트레스 조건에서 TS 함량을 유의성 있게 증가시켰고, ABMLCE와 ABMLCE + EA도 TS의 함량을 증가시켰다. TS 전구체 생성에 관여하는 HSD3B2 효소 활성은 두 조건에서 모두 EA, ABMLCE, ABMLCE + EA에 의해서 증가되었다. 또한 TS를 DTH로 전환시켜 TS함량을 감소시키는 역할을 하는 $5{\alpha}-R2$ 효소활성은 정상 조건 및 산화스트레스 조건에서 ABMLCE에 의해서만 감소되었다. Free radical로 작용하는 NO의 함량은 두 조건 모두 EA, ABMLCE, ABMLCE + EA에 의해 감소되었다. 이 결과는 EA, ABMLCE, ABMLCE + EA 처리가 TM3 세포의 HSD3B2 효소 활성을 증가시키고, NO 생성을 억제하여 TS 함량을 증가시켰음을 의미하며 EA + ABMLCE 혼합물이 남성 성기능개선 물질로 사용될 수 있음을 의미한다.

• 한국임상약학회지
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• 제22권2호
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• pp.153-159
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• 2012

Licorice is a very useful herbal medicine frequently prescribed, but glycyrrhizin, one of its components, can cause pseudoaldosteronism presenting hypokalemia, metabolic alkalosis and hypertension as a result of prolonging a biological half-life of systemic steroid by strongly inhibiting the enzyme $11{\beta}$-hydroxysteroid dehydrogenase type 2. The risk factors for this side effect are still unclear. Here, 479 hospitalized patients taking herbal medicines including licorice for more than five days were recruited to analyze their serum potassium levels, under following parameters ; age and gender of patient, dose and period of licorice, disease state such as hypertension and with or without diuretics. They were divided into the non-occurrence group (over 3.5 mEq/L) and the occurrence group (under 3.5 mEq/L) of hypokalemia based on serum potassium levels during hospitalization. The average ages of the non-occurrence group and the occurrence group were $52.6{\pm}17.8$ and $68{\pm}10.5$, respectively (p < 0.001). But there were no noticeable differences in the daily dose of licorice, days of taking, and total dose between groups. Regression analysis showed that odd ratio of age (>60) and taking diuretics inducing hypokalemia was 3.5 (95%CI 1.5-8.1), 4.6 (95%CI 2.0-10.9) that indicates significant correlation with hypokalemia. Based upon this analysis, it is imperative to monitor regularly the risk of pseudoaldosteronism or hypokalemia not only when administered licorice of high dose for long period but also when combined it with diuretics causing hypokalemia to the elderly patients over 60 year old.

• Asian-Australasian Journal of Animal Sciences
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• 제22권5호
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• pp.636-642
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• 2009

The corpus luteum (CL) is a transient endocrine gland that produces progesterone, a product required for the establishment and maintenance of pregnancy. In the absence of pregnancy, the production of progesterone in the CL decreases and the structure itself regresses in size. The life span and function of the CL are regulated by complex interactions between stimulatory (luteotrophic) and inhibitory (luteolytic) mediators. When an ovum is released from a mature follicle, angiogenesis and rapid growth of follicular cells form the CL. The purpose of the present study was to determine whether steroidogenesis, proliferation, and hypoxiarelated proteins are expressed in caprine CL. The expression of proliferating cell nuclear antigen (PCNA), vascular endothelial growth factor (VEGF), and hypoxia-inducible factor $1{\alpha}$ (HIF-$1{\alpha}$) were determined in caprine CL during the estrous cycle. Cytochrome P450 side chain cleavage protein did not vary significantly during the estrous cycle; however, there was an increased expression of $3{\beta}$ -hydroxysteroid dehydrogenase in the early and middle stages, which rapidly decreased in the late stage. The same observations were made with respect to steroidogenic acute regulatory protein. Variations in progesterone content and expression of PCNA, HIF-$1{\alpha}$, and VEGF were consistent with this result. Thus, the steroidogenic proteins, PCNA, HIF-$1{\alpha}$, and VEGF in caprine CL are dependent on the stage of the estrous cycle.

• Asian-Australasian Journal of Animal Sciences
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• 제31권5호
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• pp.658-671
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• 2018

Objective: Nandrolone decanoate (ND) is an anabolic-androgenic steroid frequently used for clinical treatment. However, the inappropriate use of ND results in the reduction of serum testosterone level and sperm production. The suppressive effect of ND on testosterone production has not been investigated in detail. The present study was designed to examine the effect of ND on the expression of steroidogenic enzymes in the rat testis. Methods: Male Sprague Dawley rats at 50 days of age were subcutaneously administrated with either 2 or 10 mg of ND/kg body weight/week for 2 or 12 weeks. The changes of transcript and protein levels of steroidogenic enzymes in the testis were determined by real-time polymerase chain reaction and western blotting analyses, respectively. Moreover, immunohistochemical analysis was employed to determine the changes of immunostaining intensity of these enzymes. The steroidogenic enzymes investigated were steroidogenic acute regulatory protein, cytochrome P450 side chain cleavage enzyme, $17{\alpha}-hydroxylase$, $3{\beta}-hydroxysteroid$ dehydrogenase, and cytochrome P450 aromatase. Results: The treatment of ND resulted in depletion of Leydig cells and sloughing of germ cells in the testis. The ND treatment caused significant expressional decreases of steroidogenic enzymes at transcript and protein levels, and the destructive effects of ND on the testis were more apparent with a higher dose and a longer period of the treatment. Evident reduction of immunostaining intensity present in Leydig cells was clearly detected by the ND treatment. Conclusion: The exposure to ND in young male results not only in histological changes of the testis but also in aberrant gene expression of testicular steroidogenic enzymes, consequently leading into the reduction of testosterone production in the testis and thus likely disruption of spermatogenesis.

• Asian-Australasian Journal of Animal Sciences
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• 제28권11호
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• pp.1573-1582
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• 2015

This study was to investigate the effect of soyabean isoflavones (SIF) on onset of puberty, serum hormone concentration, and gene expression in hypothalamus, pituitary and ovary of female Bama miniature pigs. Fifty five, 35-days old pigs were randomly assigned into 5 treatment groups consisting of 11 pigs per treatment. Results showed that dietary supplementation of varying dosage (0, 250, 500, and 1,250 mg/kg) of SIF induced puberty delay of the pigs with the age of puberty of pigs fed basal diet supplemented with 1,250 mg/kg SIF was significantly higher (p<0.05) compared to control. Supplementation of SIF or estradiol valerate (EV) reduced (p<0.05) serum gonadotrophin releasing hormone and luteinizing hormone concentration, but increased follicle-stimulating hormone concentration in pigs at 4 months of age. The expression of KiSS-1 metastasis-suppressor (KISS1), steroidogenic acute regulatory protein (StAR) and 3-beta-hydroxysteroid dehydrogenase/delta-5-delta-4 isomerase ($3{\beta}-HSD$) was reduced (p<0.01) in SIF-supplemented groups. Expression of gonadotropin-releasing hormone receptor in the pituitary of miniature pigs was reduced (p<0.05) compared to the control when exposed to 250, 1,250 mg/kg SIF and EV. Pigs on 250 mg/kg SIF and EV also showed reduced (p<0.05) expression of cytochrome P450 19A1 compared to the control. Our results indicated that dietary supplementation of SIF induced puberty delay, which may be due to down-regulation of key genes that play vital roles in the synthesis of steroid hormones.

• Asian-Australasian Journal of Animal Sciences
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• 제23권12호
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• pp.1566-1577
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• 2010

Nandrolone, 19-nortestosterone, is a synthetic androgenic-anabolic steroid promoting muscle growth. Nandrolone is also present in pig meat and sera at non-negligible levels. A number of scientific reports have suggested a positive relationship between incidence of infertility and increased meat consumption in humans. The present study was designed to determine out the effect of feeding nandrolone on the testis of the male reproductive tract. Mixtures of food and nandrolone at different concentrations (0.005 ppm and 0.5 ppm) were supplied to pubertal male rats for 6 weeks. Body weight was recorded every week during the entire experimental period. At the end of the treatment, the testis, epididymis, and epididymal fat were collected and weighted. Sperm numbers in the caudal epididymis were counted. Differential gene or protein expression of steroidogenic enzymes in the testes among experimental groups was determined by semi-quantitative real-time PCR or western blotting analysis, respectively. Histological changes of the testis induced by nandrolone treatment were examined by hematoxylin and eosin staining. Immunohistochemical analysis was employed to detect changes in the localization of steroidogenic enzymes in the testes among experimental animals. There were no significant changes on body, testis, epididymis, and epididymal fat weights among experimental groups. A significant increase of caudal sperm number was found in the 0.5 ppm nandrolone-treated group. Histological examination of the testes noted a high frequency of germ cell sloughing in seminiferous tubules of 0.5 ppm nandrolone-treated rats. Even though transcript levels of $3{\beta}$-hydroxysteroid dehydrogenase (HSD) I, $17{\beta}$-HSD4, and $17{\alpha}$-hydroxylase were influenced by nandrolone treatments, protein levels of all molecules examined in the present study were not significantly affected. Immunohistochemical analysis showed no visible changes in the localization of steroidogenic enzymes in the testes among experimental groups. The current study showed that oral intake of nandrolone in male rats for 6 weeks did not cause significant damage to the testis. It is considered that a feeding effect of nandrolone on male fertility would not be remarkable.

• 대한임상검사과학회지
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• 제51권3호
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• pp.360-369
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• 2019

Cadherin은 원형질막에 존재하며 세포-세포 결합에 관여하며, 황체 구조 유지에 필수적인 단백질이다. 본 연구에서는 prostaglandin F2 alpha ($PGF2{\alpha}$)가 황체의 협막세포(luteal theca cells, LTCs)의 E-cadherin, N-cadherin 및 세포-세포부착에 미치는 영향에 대해서 수행하였다. 황체세포는 소의 황체중기 조직으로부터 분리하였으며, 황체세포 중에서 mesenchymal 세포 형태학적 특성을 가지는 세포만을 분리하여 LTCs로 판단하였다. 이 후 steroidogenic 기능 및 혈관세포 유무를 판단하기 위해 $3{\beta}$-HSD 및 VEGF2R mRNA 발현을 확인하였으며, E-cadherin 및 N-cadherin mRNA를 사용하여 LTCs 내 cadherin의 존재여부를 판단하였다. 또한 0, $10^{-5}$, $10^{-4}$ 및 $10^{-3}M$ $PGF2{\alpha}$를 24시간 동안 처리하여 LTCs의 E- 및 N-cadherin 단백질을 관찰한 후 세포-세포 접착 실험을 실시하였다. 그 결과, LTCs에서 $3{\beta}$-HSD mRNA가 발현되었지만, VEGFR2 mRNA는 발현되지 않았으며, E-cadherin 및 N-cadherin mRNA 모두 발현되는 것을 확인하였다. 또한 E-및 N-cadherin 단백질은 $10^{-5}$, $10^{-4}$ 및 $10^{-3}M$ $PGF2{\alpha}$를 처리한 LTCs에서 응집되어 발현되는 것을 확인하였으며, $PGF2{\alpha}$에 의해 LTCs의 세포부착 효율이 유의적으로 감소된 것을 확인하였다. 결론적으로 $PGF2{\alpha}$는 LTCs의 E- 및 N-cadherin을 붕괴시켜 세포부착을 감소시켰고, 이러한 결과는 황체퇴행의 새로운 원인을 밝혀 내기 위한 cadherin과 세포부착의 역할을 이해하는데 중요한 자료로 활용될 것으로 판단된다.