• Asian-Australasian Journal of Animal Sciences
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• 제16권3호
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• pp.342-347
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• 2003

The enzyme $20{\alpha}$-hydroxysteroid dehydrogenase ($20{\alpha}$-HSD) catabolizes progesterone to $20{\alpha}$-dihydroprogesterone ($20{\alpha}$-OHP), and is appeared in rat corpora luteal and placenta. A polled samples of 10-15 placental and ovarian tissues collected from each individual rat were subjected to measurement of $20{\alpha}$-HSD activity. A $20{\alpha}$-HSD activity in the cytosol fraction was based on the generation of NADPH. In this study, it is designed to study cytosolic $20{\alpha}$-HSD activity in rat ovarian and placenta during pregnancy, and its relationship to embryonic mortality. It was found that from days 5 to 18 of pregnancy the $20{\alpha}$-HSD activities steady by decreased but at parturition time rapidly increased in ovary. On the other hand, placental cytosolic $20{\alpha}$-HSD activities were high detected from days 8 to 10 of pregnancy, not detectable from days 11 to 20 of pregnancy, but again very high at the time of parturition. Analysis of DEAE column chromatography revealed that two different types of $20{\alpha}$-HSD (HSD-1 and HSD-2) were found with similar activity in the placental cytosol on day 10 of pregnancy. The number of fetuses on day 10 of pregnancy was 15.4 and decreased significantly to 12.9 on day 12. The results suggested that expression of $20{\alpha}$-HSD in the placental tissues seems to be related the number of fetal survived in the specific time (days 11 and 12) which spontaneous fetal loss occurs.

• 대한수의학회지
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• 제34권4호
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• pp.867-872
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• 1994

Progesterone catabolizing enzyme, the enzyme $20{\alpha}$-hydroxysteroid dehydrgenase($20{\alpha}$-HSD) is pivotal in the regulation of ovarian function in rodents, which catabolizes progesterone into biologically inactive $20{\alpha}$-hydroxypregn-4-en-3-one($20{\alpha}$-OHP). In this study was carried out the influence of $20{\alpha}$-HSD activity of ovarian function, we investigated changes in ovarian cytosol $20{\alpha}$-HSD activity and serume progesterone concentration during the estrous cycles and pregnancy in rat. During the estrous cycles, the $20{\alpha}$-HSD activities were highest on the progestrous, but serum progesterone concentration was lowest on this phase. During the pregnancy, the $20{\alpha}$-HSD activities were relatively higher early pregnancy(day-1-3 gestation) and late pregnancy(day 21 to parturition), serum progesterone concentration was maintained significantly high to day 19 of gestation. The $20{\alpha}$-HSD activities were lower during the middle pregnancy. From these results, ovarian $20{\alpha}$-HSD activities may possibly act as physiologically very important in the control and maintenance of estrous cycles in rat.

• 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.

• 한국수정란이식학회지
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• 제26권1호
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• pp.57-63
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• 2011

This study was performed to the expressions of pregnancy-associated plasma protein-A (PAPP-A) and 20alpha-hydroxysteroid dehydrogenase ($20{\alpha}$-HSD) in bovine corpus luteum during early pregnancy. To determine the function of PAPP-A gene during early pregnancy, we collected corpus luteum samples on 30, 60 and 90 days of pregnancy in bovine. The mRNA expression of PAPP-A, $20{\alpha}$-HSD, progesterone-receptor (PR) and insulin-like growth factor binding protein4 (IGFBP4) gene was conducted by Real-time PCR. In parallel with mRNA levels, The protein expressions of PAPP-A and $20{\alpha}$-HSD were detected by immunological analysis. The mRNA expressions $20{\alpha}$-HSD and PAPP-A significantly increased on day 90 in the corpus luteum during pregnancy. The mRNA expression of PR and JGFBP4 in the corpus luteum progressively was enhanced at 30 to 60 day, but decreased on 90 day of pregnancy in the corpus luteum. The expression patterns of these genes, PAPP-A and $20{\alpha}$-HSD were similar pattern in these tissues. In conclusion, PAPP-A and $20{\alpha}$-HSD activity in corpus luteum could be played a role for early pregnancy manifestation.

• 대한수의학회지
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• 제41권4호
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• pp.583-589
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• 2001

To estalish bio-indices for detection of the development of multistep hepatotumorigenesis, rats were fed water containing 0.01% diethylnitrosamine (DEN) ad libitum for 13-14 weeks. Hepatocellular carcinoma was developed by treatment with DEN, DEN only was able to induce hepatic tumors in rats without any other cocarcinogen. Compared to control group, liver cytosol protein concentration in all treated grous was significantly decreased (p<0.05). From week to week, $20{\alpha}$-hydroxysteroid dehydrogenase ($20{\alpha}$-HSD) activity was increased and the highest activity was observed on the 12th week (p<0.05). In addition, the urine biopterin concentration was also significantly increased compared to control groups (p<0.05) in a time course manner. These results indicated that $20{\alpha}$-HSD activity, urine biopterin and liver cytosol protein concentration might be very useful maker to hepatic tumorigenesis.

• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 1998년도 제3차 심포지움
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• pp.10-12
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• 1998

• 대한수의학회지
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• 제34권4호
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• pp.873-880
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• 1994

Progesterone catabolizing enzyme, the enzyme $20{\alpha}$-hydroxysteroid deltydrgenase($20{\alpha}$-HSD) is pivotal in the regulation of ovarian function in rodents, which catabolizes progesterone into biologically inactive $20{\alpha}$ hydroxypregn-4-en-3-one($20{\alpha}$-OHP). In this study was carried out the influence of $20{\alpha}$-HSD activity to ovarian function and regulation such as ovulation, formation of corpus luteum, maintenance of estrous cycles and pregnancy, we investigated changes in activities of the splenic macrophages and ovarian histological findings in rat. During the estrous cycles, the ratious of phagocytotic macrophage in splenic adherent cells were highest on the proestrous, but they were lowest on metestrous. During the pregnancy, the ratious of phagocytotic macrophages in splenic adhrent cells were lowest by 1 day and then significantly increased toward the pregnancy, which were highest pregnancy on day 12. On histological findings in rat ovary, the mean number of growing, antral and mature follicles were 15.9(72.9%), 3.4(15.9%), and 2.4(11.2%), respectively. Growing follicles reached to lowest number at diestrus and mature follicle reached to lowest number at metestrus. The numbers of corpus luteums per tissue section of the ovary were 14.4 and the number of normal and atretic follicles were 11.0(76.4%) and 3.4(23.6%), respectively. The number of corpus luteum with vacolated cell were 1.7(11.8%). In this study suggests splenic macrophages are as a source of the substance which maintainer progesterone secretion from luteal cells, and are recognized pass the vessels and reside in the fresh corpora lutea soon after ovulation via inflammatory reactions, and these macrophages are felt to have a stimulatory effect on the formation of cropora lutea. In view of the results include previous report, ovarian $20{\alpha}$-hydroxysteroid dehydrogenase enzyme and splenic macrophages consider play central role in the control and maintenance of estrous cycles and pregnancy, and also applicable to both clinical and research in a wide variety such as control of reproductive system.

• Reproductive and Developmental Biology
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• 제41권1호
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• pp.1-6
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• 2017

In all mammalian species, progesterone is essential to both the preparation for, and maintenance of, pregnancy. The $20{\alpha}$-hydroxysteroid dehydrogenase ($20{\alpha}$-HSD) enzyme predominantly converts progesterone into its biologically inactive form $20{\alpha}$-hydroxyprogesterone, thereby regulating its activity. Thus, to directly assess sexual maturation in the MediKinetics $micropig^{(R)}$, we analyzed the concentration of the steroid hormones progesterone and estradiol during the estrous cycle. Our results show that the progesterone level exhibited by the analyzed $micorpig^{(R)}$ was low at the beginning of the estrous cycle, and then abruptly increased to $30.32{\pm}10.0ng/mL$ and $46.37{\pm}11.0ng/mL$ by days 9 and 11 of the cycle, respectively. It reached the highest level $55.87{\pm}3.5ng/mL$ on day 13 of the estrous cycle, before decreasing to $46.58{\pm}13.1ng/mL$ and $10.0{\pm}7.6ng/mL$ by days 15 and 17 of the cycle, respectively. In contrast, the estradiol level was shown to be highest ($27.13{\pm}11.2ng/mL$) at the initiation of the estrous cycle, after which point it decreased to $13.29{\pm}6.5ng/mL$ and $10.94{\pm}5.9ng/mL$ by days 4 and 5 of the estrous cycle, respectively. By day 17 of the estrous cycle, the estradiol level decreased to $4.13{\pm}7.6ng/mL$. We anticipate that these results will provide useful information to enable the study of human ovulation and reproductive physiology using the MediKinetics $micoripig^{(R)}$ as a model system. We recommend further investigation to elucidate the functional mechanisms underlying the regulation of sexual maturation in the MediKinetics $micropig^{(R)}$.

• Journal of Applied Biological Chemistry
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• 제59권4호
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• pp.305-311
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• 2016

본 연구는 야관문과 호로파 복합추출물인 YHM이 과산화수소($H_2O_2$)로 산화적스트레스를 가한 TM3 세포의 테스토스테론 발현에 미치는 영향을 확인하고자 수행되었다. 세포독성 시험을 수행하여 YHM의 경우 $40{\mu}g/mL$을 최고 농도로 중농도 $20{\mu}g/mL$, 저농도 $10{\mu}g/mL$ 처리군을 설정하였고, TM3 세포에 산화적 스트레스를 주기 위해서는, serum free 배지에 $50{\mu}M$의 과산화수소를 4시간 동안 처리하였다. 산화적 스트레스가 가해진 TM3 세포에 YHM 시료를 처리하여 세포 생존률에 미치는 영향을 평가하였을 때 모든 농도 처리군에서 세포증식이나 독성이 없었다. 테스토스테론은 과산화수소를 처리하였을 때 감소하였다가 YHM 시료를 처리하였을 때 control 수준으로 회복되거나, control 보다 더 증가하였다. 또한 시료에 의한 테스토스테론 양의 증가원인을 확인하기 위하여, 테스토스테론 합성 및 분해에 관여하는 효소들의 발현량을 ELISA와 Real-time PCR을 통해 알아보았다. 테스토스테론 합성에 관여하는 $3{\beta}$-HSD4와 17,20-desmorase는 과산화수소 처리 시 감소하였다가, YHM을 처리하였을 때는 control 수준으로 회복하였다. 테스토스테론을 estradiol 및 dihydrotestosterone로 변환시키는 aromatase와 $5{\alpha}$-reductase2는 과산화수소를 처리하였을 때 증가하였다가 YHM 시료를 처리하면 control 수준이나 그 이하로 감소하였다. 이 결과들로 보았을 때 YHM 시료는 TM3 세포의 증식에는 영향을 미치지 못 하지만, 산화적 스트레스에 의해 감소된 테스토스테론 합성 효소의 발현을 증가시키고, 반대로 증가되는 테스토스테론 분해 효소의 발현은 감소시켜, 결국 산화적 스트레스에 의해 저하된 테스토스테론의 양을 회복시키거나 증가시키는 효과가 있는 것으로 판단된다.