• Asian-Australasian Journal of Animal Sciences
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• v.20 no.12
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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.

• Biomedical Science Letters
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• v.19 no.3
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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.

• Korean Journal of Animal Reproduction
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• v.16 no.2
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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.

• Journal of Animal Reproduction and Biotechnology
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• v.36 no.3
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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.

• Journal of Life Science
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• v.28 no.6
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• pp.648-655
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• 2018

Enhancement mechanistic actions of testosterone (TS) productions in mouse Leydig TM3 cells by the eritadenine (EA) and/or the Agaricus blazei mycelial liquid culture extract (ABMLCE). Productions of TS in TM3 cells were investigated in normal and oxidative-stressed culture conditions. In the normal culture condition, TM3 cells grown in a Dulbecco's Modified Eagle's Medium (DMEM) were treated with EA (0~100 ppm) and ABMLCE (10 ppm) + EA (0~50 ppm) for 24 hr, and in the oxidative-stressed culture condition, the cells grown in DMEM containing $50{\mu}M$ $H_2O_2$ to induce oxidative stress for 4 h were treated with the same as those in the normal culture condition. TS content, $3{\beta}$-hydroxysteroid dehydrogenase 2 (HSD3B2) enzyme activity, $5{\alpha}$-reductase 2 ($5{\alpha}-R2$) enzyme activity, and free-radical nitric oxide (NO) content in the culture media were measured using their corresponding assay kits. EA, ABMLCE, and ABMLCE + EA significantly, p<0.05, enhanced TS productions in both cultural conditions, relative to control treatment. The activity of the HSD3B2 enzyme, which is involved in the production of precursors for TS production, was elevated by EA, ABMLCE, and ABMLCE + EA treatments in both culture conditions. The activity of the $5{\alpha}-R2$ enzyme, which converts TS to dihydroxytestosterone (DHT), was not significantly affected in either culture condition by EA, ABMLCE, or ABMLCE + EA treatments. The treatments included reduced NO content. These results indicate that EA, ABMLCE, and EA + ABMLCE treatments elevated TS in TM3 cells via the enhancements of HSD3B2 activity and the reduction of NO production, and also imply that EA and ABMLCE or EA + ABMLCE could be useful materials for the production of TS in humans.

• Asian-Australasian Journal of Animal Sciences
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• v.22 no.5
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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.

• Korean Journal of Animal Reproduction
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• v.16 no.2
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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.

• Korean Journal of Animal Reproduction
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• v.16 no.2
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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.

• Korean Journal of Animal Reproduction
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• v.19 no.4
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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.

• Journal of Nutrition and Health
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• v.50 no.3
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• pp.217-224
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• 2017

Purpose: Although it is well known thatmortality and morbidity due to cardiovascular diseases are higher in salt-sensitive subjects than in salt-resistant subjects, their underlying mechanisms related to obesity remain unclear. Here, we focused on salt-sensitive gene variants unrelated to monogenic obesity that interacted with sodium intake in humans. Methods: This review was written based on the modified $3^rd$ step of Khans' systematic review. Instead of the literature, subject genes were based on candidate genes screened from our preliminary Genome-Wide Association Study (GWAS). Finally, literature related to five genes strongly associated with salt sensitivity were analyzed to elucidate the mechanism of obesity. Results: Salt sensitivity is a measure of how blood pressure responds to salt intake, and people are either salt-sensitive or salt-resistant. Otherwise, dietary sodium restriction may not be beneficial for everyone since salt sensitivity may be associated with inherited susceptibility. According to our previous GWAS studies, 10 candidate genes and 11 single nucleotide polymorphisms (SNPs) associated with salt sensitivity were suggested, including angiotensin converting enzyme (ACE), ${\alpha}$-adducin1 (ADD1), angiotensinogen (AGT), cytochrome P450 family 11-subfamily ${\beta}$-2 ($CYP11{\beta}$-2), epithelial sodium channel (ENaC), G-protein b3 subunit (GNB3), G protein-coupled receptor kinases type 4 (GRK4 A142V, GRK4 A486V), $11{\beta}$-hydroxysteroid dehydrogenase type-2 (HSD $11{\beta}$-2), neural precursor cell-expressed developmentally down regulated 4 like (NEDD4L),and solute carrier family 12(sodium/chloride transporters)-member 3 (SLC 12A3). We found that polymorphisms of salt-sensitive genes such as ACE, $CYP11{\beta}$-2, GRK4, SLC12A3, and GNB3 may be positively associated with human obesity. Conclusion: Despite gender, ethnic, and age differences in genetics studies, hypertensive obese children and adults who are carriers of specific salt-sensitive genes are recommended to reduce their sodium intake. We believe that our findings can contribute to the prevention of early-onset of chronic diseases in obese children by facilitating personalized diet-management of obesity from childhood to adulthood.