• Asian-Australasian Journal of Animal Sciences
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• 제20권12호
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• pp.1832-1842
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• 2007

Cytochrome P450 aromatase is responsible for the biosynthesis of estrogen. It is also responsible for the endogenous production of 19-nortestosterone (nandrolone), an anabolic androgen unique to pigs. Plasma concentrations of 19-nortestosterone are highest between two and four weeks after birth in male pigs. In the present study, the physiology of 19-nortestosterone was investigated by measuring the mRNA levels of steroidogenic enzymes, estrogen receptors and androgen receptor in the tissues of growing pigs. The expression of aromatase, 17${\alpha}$-hydroxylase and 3${\beta}$-hydroxysteroid dehydrogenase in the testes of male piglets increased between birth and two weeks of age, and then decreased progressively. Similar developmental expressional patterns were observed for 17${\alpha}$-hydroxylase and 3${\beta}$-hydroxysteroid dehydrogenase in the ovaries of female piglets, but without significant aromatase expression. The major form of aromatase expressed in the testes of piglets was identified as type I. Expression of estrogen receptor-${\alpha}$ and -${\beta}$and androgen receptor genes was also detected in both testes and ovaries. A transient elevation of androgen receptor mRNA in male piglets at two weeks of age was also observed in testes. Significant expression of the androgen receptor gene, but not of estrogen receptor-${\alpha}$ and -${\beta}$ genes, was also demonstrated in adipose tissue and muscle. We conclude that the observed increase in the testicular expression of aromatase in male pigs could account for the production of large amounts of 19-nortestosterone at between two and four weeks of age in males. Androgen receptor and 19-nortestosterone appeared to be important for testicular development and might contribute to sexual dimorphism in body composition and muscle development in juvenile pigs.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2007년도 Proceedings of The Convention
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• pp.57-64
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• 2007

Metabolic interactions of the three major cannabinoids, ${\Delta}^9$-tetrahydrocannabinol (THC), cannabidiol (CBD), and cannabinol (CBN) with steroid hormones were investigated. These cannabioids concentration-dependently inhibited $3{\beta}$-hydroxysteroid dehydrogenase and $17{\alpha}$-hydroxylase in rat adrenal and testis microsomes. CBD and CBN were the most potent inhibitors of $3{\beta}$-phydroxysteroid dehydrogenase and progesterone $17{\alpha}$-hydroxylase, respectively, in rat testis microsomes. Three cannabinoids highly attenuated hCG-stimulated testosterone production in rat testicular interstitial cells. These cannabinoids also decreased in levels of mRNA and protein of StAR in the rat testis cells. These results indicate that the cannabinoids could interact with steroid hormones, and exert their modulatory effects on endocrine and testicular functions. Metabolic interaction of a THC metabolite, $7{\beta}$-hydroxy-${\Delta}^8$-THC with steroids is also investigated. Monkey liver microsomes catalyzed the stereoselective oxidation of $7{\beta}$-hydroxy-${\Delta}^8$-THC to 7-oxo-${\Delta}^8$-THC, so-called microsomal alcohol oxygenase (MALCO). The reaction is catalyzed by CYP3A8 in the monkey liver microsomes, and required NADH as well as NADPH as an efficient cofactor, and its activity is stimulated by some steroids such as testosterone and progesterone. Kinetic analyses revealed that MALCO-catalyze reaction showed positive cooperativity. In order to explain the metabolic interaction between the cannabinoid metabolite and testosterone, we propose a novel kinetic model involving at least three binding sites for mechanism of the metabolic interactions.

• 대한의생명과학회지
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• 제19권3호
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• pp.261-265
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• 2013

The corpus luteum is a transient endocrine gland essential for regulation of the ovarian cycle as well as for establishing and maintaining pregnancy. Prostaglandin $F_{2{\alpha}}$ (PGF) initiates functional and structural regression of the corpus luteum and therefore is an important regulator of the estrous cycle. It is a matter of debate whether the endothelial cells of the bovine corpus luteum express PGFR, the cognate receptor for PGF. Therefore, the aim of this study was to assess the expression of PGFR in bovine endothelial cells. Endothelial cells were isolated from the bovine corpus luteum of the mid-luteal stage using magnetic beads and cultured in vitro. We demonstrate that this isolation procedure generates a pure culture of endothelial cells as confirmed by synthesis of Factor VIII and lack of expression of $3{\beta}$-hydroxysteroid dehydrogenase. By RT-PCR, Western blot and immunofluorescence analyses, we further show that the cultured endothelial cells produced PGFR. This model system can be utilized to provide an experimental system to investigate the role of PGF on endothelial cells during the reproductive cycle.

• 한국가축번식학회지
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• 제16권2호
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• pp.133-139
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• 1992

The study was conducted to find out the changes of progesterone and 20$\alpha$-dihydroprogesterone (20$\alpha$-OHP) levels during the oestrous cycle in Korean native goats. 4 cyclic goats were offered for this experiment. Blood samples were taken from jugular vein on day 0, 1, 3, 5, 7, 9, 11, 13, 15, 17 and 19 during the oestrous cycle, then the next oestrous day. The serum levels of progesterone and 20$\alpha$-OHP were measured by radioimmunoassay. The progesterone concentration in seurm of the cyclic goats was 0.29$\pm$0.06ng/ml on the first day of oestrous (day 0), increased to 5.29$\pm$0.73ng/ml on day 9, reached to a peak level of 5.73$\pm$0.61ng/ml on day 13, and thereafter decreased to 0.35$\pm$0.30ng/ml on day 1. The serum level of 20$\alpha$-OHP during the oestrous cycle was 0.42$\pm$0.33ng/ml on day 0 and then decreased to 0.28$\pm$0.01ng/ml on day 5. This basal level was maintained until day 13, increased gradually, and reached a peak level of 0.62$\pm$0.05ng/ml on day 19. From the above results, it was suggested that the enzyme 20$\alpha$-hydroxysteroid dehydrogenase(20$\alpha$-HSD) catalyzing the conversion of progesterone to a biologically inactive steroid, 20$\alpha$-OHP, should be active in the luteal cells during the oestrous cycle in Korean native goats.

• 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.

• 한국가축번식학회지
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• 제16권2호
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• pp.149-155
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• 1992

The present study was conducted to find out the changes of progesterone and 20$\alpha$-dihydroprogesterone(20$\alpha$-OHP) levels before and after parturition, 4 pluriparous goats were offered for this experiment. Blood samples were taken from jugular vein on Days, 5, 3, 2 and 1 before parturition, the day of parturition, 1, 3, 5, 7 and 9 after parturition. The blood samples were centraifuged and stored at -2$0^{\circ}C$ until hormone assay. The serum levels of progesterone and 20$\alpha$-OHP were measrued by radioimmunoassay. The changes of serum progesterone level during peripartum period were characterized as a remarkable decrease. The progesterone level was 4.05$\pm$0.52ng/ml on 56 days before parturition and decreased to 2.24$\pm$0.38ng/ml on 1 day before parturition and 0.79$\pm$0.09ng/ml on the day of parturition and the basal level was maintained through 9 days of postpartum period. The serum level of 20$\alpha$-OHP during the peripartum period was 1.25$\pm$0.21ng/ml on 5 days before paturition and increased to 1.32$\pm$0.25 on 3 days and 1.59$\pm$0.24ng/ml on 1 day before parturition, and reached a peak level of 1.78$\pm$0.25ng/ml just prior to parturition and then decreased greatly to 0.31$\pm$0.03ng/ml on 1 day postpartum and the basal level was remained until 9 days postpartum. The high serum level of 20$\alpha$-OHP, which was peak just prior to parturition, was maintained for 2 days following the onset of remarkable decrease in the serum level of progesterone. From the above results, it was concluded that the enzyme 20$\alpha$-hydroxysteroid dehydrogenase (20$\alpha$-HSD) catalyzing the conversion of progesterone to a biologically inactive steroid, 20$\alpha$-OHP was active properly in the luteal cells in Korean native goats.

• 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.

• 한국동물생명공학회지
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• 제36권3호
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• pp.137-144
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• 2021

In this study, to analyze whether artificial regulation of apoptosis in the development of somatic cells can affect the stable growth and development of cells, 20 alpha-hydroxysteroid dehydrogenase (20α-HSD) and rapamycin were treated to induce apoptosis and autophagy in the both skin and muscle cells. Respectively, and 3-methyladenine was supplemented to inhibit cell death. Our results show that stimulation with rapamycin activated autophagy in both types of cells, but increased apoptosis more than autophagy in the case of skin cells. These results indicate that there was a difference in the expression of survival factors and cell development depending on the type of cell. In particular, in the expression of autophagy-related gene (MAP1LC3A) was higher than that of Casp-3, an apoptosis factor. Furthermore, cell development was the highest in all cell groups cultured by artificially inducing autophagy, however the lowest in the apoptosis-inhibited group. Especially, the noteworthy result of this study was that when apoptosis was induced using 20α-HSD, it was possible to induce apoptosis in both skin and muscle cells. Therefore, the main point of this study is that apoptosis induced during cell culture plays a pivotal role in cell remodeling.

• 한국가축번식학회지
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• 제16권2호
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• pp.141-147
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• 1992

The present study was conducted to find out the changes of progesterone and 20$\alpha$-dihydroprogesterone(20$\alpha$-OHP) levels during the gestation period in Korean native goats. 4 pregnant goats were offered for this experiment. Blood samples were taken from jugular vein on Day 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140 and 145 of the pregnant goats. The serum levels of progesterone and 20$\alpha$-OHP were measured by radioimmunoassay. The progesterone level in goat serum during the gestaton was low as 2.94$\pm$0.21 at 5 days, and then increased to 4.51$\pm$0.25ng/ml at 10 days of gestation and increased greatly from Days 60 and reached a peak level of 7.21$\pm$0.58ng/ml at 100 days of gestation, and thereafter decreased to 6.01$\pm$0.57, 5.26$\pm$0.64ng/ml on Days 130, 140 of gestation, and decreased to 4.05$\pm$0.52ng/ml on Days 145 of pregnancy. The serum level of 20$\alpha$-OHP during the gestation was low as 0.3~0.4ng/ml in the early stage of pregnancy, and increased gradually and increased to 0.85$\pm$0.06ng/ml, 0.97$\pm$0.08ng/ml on Days 90 or 100, and then increased to 1.18$\pm$0.18, 1.25$\pm$0.21ng/ml on Days 140 or 145 of gestation. While the serum levels of progesterone during the luteal regression decreased, the 20$\alpha$-OHP increased continuously. From the above results, it was concluded that the enzyme 20$\alpha$-hydroxysteroid dehydrogenase (20$\alpha$-HSD) catalyzing theconversion of progesterone to a biologically inactive steroid, 20$\alpha$-OHP was active properly in the luteal cells what the levels of progesterone decreased and the levels of 20$\alpha$-OHP increased during the late pregnancy in Korean native goats.

• 한국가축번식학회지
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• 제19권4호
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• pp.307-322
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• 1996

The corpus luteum (CL) is formed by the action of a surge of luteinizing hormone (LH) on the pre-ovulatory follicle. Luteal cells derived from granulosa and theca interna cells continue to secrete progesterone for about two weeks. LH in domestic animals is essential for the normal secretion of progesterone at all stages of the luteal phase. For this process in the rodents, 20$\alpha$-hydroxysteroid dehydrogenase (20$\alpha$-HSD) is indispensable. 20$\alpha$-HSD is an enzyme to be a biologically inactive steroid. This enzyme plays a critical role in the regulation of the rat luteal function and reported to be present in steroid-producing tissues such as the testis and adrenal gland. We have purified 20$\alpha$-HSD and found two distinct 20$\alpha$-HSD molecules (HSD-1 and HSD-2). Their molecular weights are both estimated to be 33kd.The amino acid compositions of HSD-1 and HSD-2 are mostly similar, but there is a slight difference in the content of lysine. We demonstrated that 1) CL of previous generations contribute more to whole ovarian 20$\alpha$-HSD activity, 2) newly formed corpora lutea contain only 20$\alpha$-HSD-1 activity, and 3) old CL express activities of each HSD isozyme as shown in the luteal tissue of cycling rats on the day of diestrus where only degenerating old CL exist. The increase in 20$\alpha$-HSD activity identified seems to be related to the increase in the numbers of 20$\alpha$-HSD-positive cells. Interestingly, 20$\alpha$-HSD-1 activities were strongly found in the follicle fluids and theca interna cells by immunohistochemical study. Thus, the activity of 20$\alpha$-HSD may be related to a survival mechanism of those luteal cells and follicles remaining in the ovaries. Luteal cells arise from two sources. The small luteal cells are all of theca cell origin, while the large luteal cells are mainly of granulosa cell origin. CL of Korean Native Cattle, as those of other animal species, contains two morphologycally and functionally distinct luteal cell populations, such as small and large luteal cells as well as nonluteal cells. In all reproductive states except in the late luteal phase, the bovine CL also contained more small luteal cells than large luteal cells. Luteal tissue secretes a variety of growth factors (proteins) and the pattern of secretion changes during all stages of the luteal phase. These growth factors could be important in regulating the function of the bovine corpus luteum and may act in a potential endocrine autocrine and paracrine mechanisms. Therefore, further work has to be done to elucidate the role of growth factors in the ovary, especially in the corpus luterum. Interest should be focussed on interaction of these growth factors in the regulation of luteal cell and the localization of cytokine synthesis in differnet luteal cells.