• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.82-82
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• 2001

본 연구는 한우에 있어서 인공수정후 수태율증진을 위한 기초자료를 확보할 목적으로 성숙 미경산 한우의 발정동기화를 위해 CIDR(EAZI-BREED CIDR Plus, USA)를 7일간 질내에 삽입하였고, CIDR처리 제 6일째에 PGF2$\alpha$(lutlyase, USA)제제 5 $m\ell$을 근육주사하여 발정이 유기된 92%의 개체를 인공수정을 실시하였다. 수태율증진을 위해 무처리구를 두었고, hCG처리구(Chorulon, Intervet, Netherlands)는 2000IU를 근육 주사하였으며, CIDR처리구에는 Estradiol-17$\beta$의 캡슐을 제거한 CIDR를 수정후 7일부터 일주일간 질내에 삽입 처리하여 각 처리당 15두씩 총 45두의 한우를 실험에 공시하였다. 발정이 유기된 개체는 인공수정후 30일까지 2일 간격으로 채혈하여 혈중 Progesterone, IGF, IGF-II 및 cortisol 농도를 RIA법으로 분석하였으며 수정 후 60일 이후에는 직장검사법으로 임신감정을 실시하였다. 수정후 수태율은 대조구와 CIDR 처리구에서는 평균 62%였으나 hOG처리구는 73%로서 대조구에 비해 유의적으로 수태율이 높게 나타났다. 이때의 혈중 progesterone농도를 분석한 결과, 대조구에 CIDR 처리구의 수정후 7일째부터 증가하기 시작하여 제 12일째에서 평균 5.5ng/$m\ell$까지 증가하다가 그 이후 재발정 예정일일 21일째에 다소 감소되다가 다시 증가되어 일정한 수준으로 유지되었다 그러나 hCG처리구의 경우 수정후 5일부터 progesterone이 증가하다가 제 15일까지 약 10ng/$m\ell$까지 높은 progesterone이 유지되었다. 이와 같은 결과로 보아 hCG는 혈중 progesterone농도를 증가시킴으로써 난소내 임신황체의 progestorone분비 능을 촉진시키는 작용을 하는 것으로 사료되나, 한편 혈중 IGF I과 IGF-II농도는 대조구, hCG처리구 및 CIDR 처리구간의 차이가 없이 수정 후 제10일까지는 다소 높은 수준이었으나 그 이후 감소되는 현상으로 혈중 progesterone농도와는 부의 상관관계를 보였다. 따라서 IGF-I과 IGF-II는 혈중 progesterone의 농도를 인위적으로 조절하거나 황체의 progesterone분비기능을 직접 조절하는 역할은 하지 않는 것으로 사료되며, 특히 초기 임신의 수태율에 직접적으로 영향이 없는 것으로 사료된다. 또한 혈중 cortisol농도는 처리간의 차이는 확인할 수 없으나 임신과 비임신간의 혈중 cortisol농도는 유의적으로 차이가 있는 것으로 나타났다. 본 실험의 결과로 인공수정이나 수정란이식 및 체세포복제 수정란뿐만 아니라 형질전환 수정란의 이식후 수태율을 증진시켜 첨단기술의 조기정착을 위해 이식 후 7일째나 인공수정 후 7일째 hCG제제를 근육주사로 간단하게 처리함으로써 수태율을 크게 개선될 수 있으며 이것은 난소의 황체기능 즉, 황체로부터 충분한 progesterone을 분비할 수 있는 내분비적 환경을 제공하는 것으로 사료된다.

• Reproductive and Developmental Biology
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• v.32 no.2
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• pp.117-121
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• 2008

Our previous study showed that transgenic (TG) pigs harboring human EPO (hEPO) gene have been shown to have reproductive disorders, including low pregnancy rates, irregular estrus cycle and low little size. To investigate these reasons, we assessed estrus behavior (standing response) and plasma $17{\beta}$-estradiol ($E_2$) level, which partly reflect reproductive function, during the estrus cycles after synchronization and superovulation by hormone treatments. Then, we analysed blood composition and expression of hEPO gene in TG pigs. Pigs were injected with PG600. After 10 days, pigs were fed with Regumate porcine for 6 days. Blood samples were collected from jugular vein. Analysis of blood composition and $E_2$ level were measured by Hemavet 950 and $E_2$ ELISA kit, respectively. And, the expression of hEPO gene in reproductive organs was quantitated by real-time RT-PCR. The percentage of estrus behavior in TG was significantly decreased. Hematocrit (HCT), hemoglobin (Hb) concentration and red blood cell (RBC) number were significantly higher in TG than wild type (WT). On the other hand, high expression of hEPO gene in TG was observed in the mammary gland as well as in the uterus. Moreover, plasma $E_2$ level was significantly higher in TG than WT. These results suggest that nonspecific expression of hEPO gene in the other organs of TG may affect blood composition and plasma $E_2$ level, thereby causing reproductive disorders.

• Clinical and Experimental Reproductive Medicine
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• v.25 no.3
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• pp.251-260
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• 1998

It is well known that the clinical test for responsibility of accurate fertilization capacity in male partners is very important to diagnose and treat the infertility. However, it has been reported that the traditional semen analysis cannot accurately predict fertilization and pregnancy potential. The present study was performed to evaluate the acrosomal reaction to ionophore challenge (ARIC) test as a prognostic indicator for fertilization of sperm and oocyte in an in vitro fertilization and embryo transfer (IVF-ET) program. From March 1996 to Februry 1997, 30 couples undergoing IVF program were allocated to this study group. All female partners in the study group were 35 years old or less and their serum level of basal follicle stimulating hormone (FSH) and estradiol $(E_2)$ were normal. All the male partners have normal parameters of semen analysis. The ARIC tests were performed on the day of ovum pick up and in vitro insemination in all the male partners. The controlled ovarian hyperstimulation (COH) using luteal long protocol of gonadotropin releasing hormone (GnRH) agonist was used in all couples for IVF-ET. The acrosomal reaction with $10{\mu}l$ of 10% DMSO was induced spontaneously in $10.1{\pm}9.8%$, and acrosomal reaction with calcium ionophore A 23187 was induced in $27.4{\pm}18.1%$, and the ARIC value was $17.4{\pm}16.2%$. There were no significant correlation between the ARIC value and the fertilization rate ($r^2$=0.044, p=0.268). There were also no significant correlation between the ARIC value and the percentage of the grade I, II embryos ($r^2$=0.046, p=0.261). On the basis of above results, it was suggested that ARIC test might not be a useful prognostic indicator for fertilization in IVF-ET in male partners with normal parameters of conventional semen analysis. We guessed that IVF-ET could be performed to the patients primarily without universal appilcation of ARIC test to all male partenrs, and if fertilization failure occurs, the micro assisted fertilization (MAF) such as intracytoplsmic sperm injection (ICSI) might be used as an alternative mode of treatment with acceptable success rate.

• Asian-Australasian Journal of Animal Sciences
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• v.25 no.1
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• pp.44-51
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• 2012

Calcium ions play an important role in the establishment and maintenance of pregnancy, but molecular and cellular regulatory mechanisms of calcium ion action in the uterine endometrium are not fully understood in pigs. Previously, we have shown that calcium regulatory molecules, transient receptor potential vanilloid type 5 (TRPV6) and calbindin-D9k (S100G), are expressed in the uterine endometrium during the estrous cycle and pregnancy in a pregnancy status- and stage-specific manner, and that estrogen of conceptus origin increases endometrial TRPV6 expression. However, regulation of S100G expression in the uterine endometrium and conceptus expression of S100G has been not determined during early pregnancy. Thus, we investigated regulation of S100G expression by estrogen and interleukin-$1{\beta}$ (IL1B) in the uterine endometrium and conceptus expression of S100G during early pregnancy in pigs. We obtained uterine endometrial tissues from day (D) 12 of the estrous cycle and treated with combinations of steroid hormones, estradiol-$17{\beta}$ ($E_2$) and progesterone ($P_4$), and increasing doses of IL1B. Real-time RT-PCR analysis showed that $E_2$ and IL1B increased S100G mRNA levels in the uterine endometrium, and conceptuses expressed S100G mRNA during early pregnancy, as determined by RT-PCR analysis. To determine if endometrial expression of S100G mRNA during the implantation period was affected by the somatic cell nuclear transfer (SCNT) procedure, we compared S100G mRNA levels in the uterine endometrium from gilts with SCNT-derived conceptuses with those from gilts with conceptuses derived from natural mating on D12 of pregnancy. Real-time RT-PCR analysis showed that levels of S100G mRNA in the uterine endometrium from gilts carrying SCNT-derived conceptuses was significantly lower than those from gilts carrying conceptuses derived from natural mating. These results showed that S100G expression in the uterine endometrium was regulated by estrogen and IL1B of conceptus origin, and affected by the SCNT procedure during early pregnancy. These suggest that conceptus signals regulate S100G, an intracellular calcium transport protein, for the establishment of pregnancy in pigs.

• Asian Pacific Journal of Cancer Prevention
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• v.15 no.1
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• pp.495-499
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• 2014

Estrogens are considered the major breast cancer risk factor, and the carcinogenic potential of estrogens might be attributed to DNA modification caused by derivatives formed during metabolism. $17{\beta}$-estradiol ($E_2$), the main steroidal estrogen present in women, is metabolized via two major pathways: formation of 2-hydroxyestradiol (2-OH $E_2$) and 4-hydroxyestradiol ($4-OH\;E_2$) through the action of cytochrome P450 (CYP) 1A1 and 1B1, respectively. Previous reports suggested that $2-OH\;E_2$ has putative protective effects, while $4-OH\;E_2$ is genotoxic and has potent carcinogenic activity. Thus, the ratio of $2-OH\;E_2/4-OH\;E_2$ is a critical determinant of the toxicity of $E_2$ in mammary cells. In the present study, we investigated the effects of berberine on the expression profile of the estrogen metabolizing enzymes CYP1A1 and CYP1B1 in breast cancer MCF-7 cells. Berberine treatment produced significant induction of both forms at the level of mRNA expression, but with increased doses produced 16~ to 52~fold greater induction of CYP1A1 mRNA over CYP1B1 mRNA. Furthermore, berberine dramatically increased CYP1A1 protein levels but did not influence CYP1B1 protein levels in MCF-7 cells. In conclusion, we present the first report to show that berberine may provide protection against breast cancer by altering the ratio of CYP1A1/CYP1B1, could redirect $E_2$ metabolism in a more protective pathway in breast cancer MCF-7 cells.

• The Korean Journal of Physiology
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• v.21 no.1
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• pp.35-46
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• 1987

It has been well known that estrogens stimulate the uterine contractility and progestins inhibit it. Then, one may expect that the uterine contractility and sensitivities to oxytocin (OT) and prostaglandin $F_{2{\alpha}}\;(PGF_{2{\alpha}})$ would be different among the estrus cycle. These hypotheses were tested using the mature female rat. Spontaneous isometric contractions of isolated uterine strips $(1{\times}0.3\;cm)$ from cyclic rats in various stages of the estrus cycle, bilateral ovarectomized rats and hypophysectomized rats were recorded in absence or presence with $estradiol-17{\beta}\;(E_2)$, progesterone $(P_4)$, OT and $PGF_{2{\alpha}}$. The results were summarized as follows: 1) The spontaneous uterine contractile force was the highest in the estrus rat and the lowest in the ovarectomized or the hypophysectomized rat. In the proestrus rat, the contractile frequency was the lowest (2.7 beats/10 min) and the contractile duration was the longest (70 sec). In the other groups, there were no any differencies in frequency (9 beats/10 min) and in duration (30 sec). 2) OT and $PGF_{2{\alpha}}$ stimulated the uterine contractility in all groups tested except in the hypophysectomized rat in which OT failed to stimulate the uterine contraction. $PGF_{2{\alpha}}$ was more effective in stimulating the uterine contraction than OT in all groups tested except in the estrus rat. OT-induced contraction was the highest in the estrus rat and $PGF_{2{\alpha}}-induced$ contraction was the lowest in the hypophysectomized rat. 3) Uterine contractilities were not changed by the in vitro treatments of $E_2$ or $P_4$ under the influence of endogenous steroids, however, $E_2$ and $P_4$ stimulated the uterine contraction in the ovarectomized rat in which endogenous steroids were almost abolished. 4) Increased uterine contraction by the treatment of OT was suppressed by in vitro $E_2$ or $P_4$ in the estrus rat, while it was potentiated by the $P_4$ in the proestrus rat. In other groups, exogenous $E_2$ or $P_4$ did not affect the OT-induced uterine contraction. 5) $PGF_{2{\alpha}}-induced$ uterine contraction was suppressed in the ovarectomized rat by $E_2$ and $P_4$, in the diestrus and proestrus rats by $P_4$ and in the hypophysectomized rat by $E_2$. In other groups, exogenous $E_2$ or $P_4$ was ineffective in altering the $PGF_{2{\alpha}}-induced$ uterine contraction. According to the above results, it may conclude that the mechanisms of the different uterine contractility and the different uterine sensitivity to OT or $PGF_{2{\alpha}}$ according to the estrus cycle are not explicable with only the serum concentrations of steroids, OT and $PGF_{2{\alpha}}$ but also other unknown factors.

• The Journal of Korean Obstetrics and Gynecology
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• v.24 no.3
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• pp.28-47
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• 2011

Objectives: This study was performed to investigate the effects of Angelicae gigantis Radix (AGR) which is one of the most useful herbal-drug to treat patients with Polycystic Ovary Syndrome (PCOS) in Oriental medicine on gene expression of ovary tissue. Methods: The effects of AGR on gene expression of ovary tissue resected from PCOS induced rats using single injection of $\ss$-Estradiol 17-valerate (EV) was measured using microarray technique, and the functional analysis on these genes was conducted. Results: Total 2,812 genes were up-regulated or down-regulated, 1,421 genes were up-regulated, 1,391 genes were down-regulated by induction of PCOS. Up-regulated genes were mainly involved in biological function such as cell signalling pathways and inflammatory response. Expression levels of 1,442 genes were restored to those of naive animals by administration of AGR. 558 genes were restored to those of naive animals, which were lowered by induction of PCOS. 884 genes were lowered to naive levels, which were elevated by induction of PCOS. The functions of restored genes were partially involved in the restoration of expression levels, which were changed by induction of PCOS. Especially, up-regulated gene by induction of PCOS were mainly involved in these changes. These results mean restorative effects of AGR on damaged functions by induction of PCOS. The network of total protein interactions was measured using cytoscape program, and some key molecules, such as IRS2, MCM10, ORC2L related in up-regulated genes, CTBP2, CD44, RHOA, related in down-regulated genes that can be used for elucidation of therapeutical mechanism of medicine in future were identified. Conclusion: Restored genes by AGR were thought to have common pathways related in regulation of gene expressions. Especially, genes in restored expression levels by AGR, which were up-regulated by induction of PCOS, were regulated by 9 of common transcription factors, genes in restored expression levels by AGR, which were down-regulated by induction of PCOS, were involved in 25 of common transcription factors.

• Proceedings of the Korean Society of Postharvest Science and Technology of Agricultural Products Conference
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• 2003.04a
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• pp.22-39
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• 2003

Biological activity of phenolic compounds in seeds and leaves of safflower (Carthamu tinctorius L.) were evaluated using several in vitro and in vivo assays. Six phenolic constituents were isolated from the seeds and identified as N-feruloylserotonia, N- (p-coumaroyl)serotonin, matairesinol, 8′-hydroxyarctigenin, acacetin 7-O-$\beta$-D-glucoside (tilianine) and acacetin. Six phenolic compounds exhibited considerable antioxidative activity, and especially two serotonins showed potent DPPH radical scavenging activity and antiperoxidative activity against rat liver microsomal lipid peroxidation induced by the hydroxyl radical generated via a Fenton-type reaction. Additionally, six phenolic compounds possessed comparable cytotoxicity against three cancer cells, Hela cell, MCF-7 and HepG2 cell, and particularly acacetin and its glycosides had the most potent cytotoxicity. Moreover, we found that feeding safflower seeds attenuated bone loss, and lowered levels of plasma and liver lipids in ovariectomized rats. Serotonins, lignans and flavones stimulated proliferation of the osteoblast-like cells in a dose-dependent manner (10$^{-15}$ ~10$^{-6}$ M), as potently as E$_2$ (17$\beta$-estradiol). Particularly, serotonins were mainly responsible for bone-protecting and lipid lowering effects in ovariectomized rats. Meanwhile, eight flavonoids, including a novel quercetin-7-O-(6"-O-acetyl)-$\beta$-D-glucopyranoside and seven kown flavonoids, luteolin quercetin, luteolin 7-O-$\beta$-D-glucopyranoside, luteolin-7-O-(6"-O-acetyl)-$\beta$-D-gluco-pyranoside, quercetin 7-O- -glucopyranoside, acacetin 7-O-$\beta$-D-glucuronide and apigenin-6-C-$\beta$-D-glucopyranosyl-8-C-$\beta$-D-glucopyranoside were first isolated and identified from safflower leaf. Among these flavonoids, luteolin-acetyl-glucoside and $\beta$quercetin- acetyl-glucoside showed potent antioxidative activities against 2-deoxyribose degradation and lipid peroxidation in rat liver microsomes. Luteolin, quercetin and their corresponding glycosides also exhibited strong antioxidative activity, while acacetin glucuronide and apigenin-6, 8-di-C-glucoside were relatively less active. Finally, changes in phenolic compositions were also determined by HPLC in the safflower seed and leaf during growth stages and roasting process to produce standardized supplement powerds. These results suggest that phenolic compounds in the roasted safflower seed and leaf may be useful as potential sources of therapeutic agents against several pathological disorders such as carcinogenesis, atherosclerosis and osteoporosis.

• Korean Journal of Food Science and Technology
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• v.32 no.4
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• pp.964-969
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• 2000

Di-(2-ethylhexyl) adipate(DEHA) has been used extensively as a plasticizer in the manufacture of plastic products such as PVC films. Though, phthalate esters plasticizers have been known to induce endocrine system-mediated responses, few studies have been conducted for the screening of estrogenic activity of DEHA, an adipate plasticizer. This study was initiated to evaluate the estrogenic activity of DEHA by in vitro E-screen assay and in vivo uterotrophic assay. MCF-7 human breast cancer cells were treated with $DEHA(5{\times}10^{-9}{\sim}5{\times}10^{-4}\;M)$, for 144 hr, and cell proliferation was determined by sulforhodamine B(SRB) assay. DEHA dissolved in corn oil was administered subcutaneously to ovariectomized(OVX) female Sprague-Dawley rats at dosage levels of 0, 2, 20 and 200 mg/kg/day for three consecutive days. Rats were sacrificed 24 hr after final treatment and vagina and uterus(wet and blotted) weights were obtained. E-screen assayed DEHA did not generate cell proliferation at treated concentrations$(5{\times}10^{-9}{\sim}5{\times}10^{-4}\;M)$, whereas 17 ${\beta}-estradiol$(E2), the positive control, induced cell proliferation at low concentrations$(5{\times}10^{-14}{\sim}5{\times}10^{-9}\;M)$. In the uterotrophic assay, DEHA did not change vagina and uterus(wet and blotted) weights at dosage levels up to 200 mg/kg/day treatment. These results demonstrated that DEHA did not exhibit the estrogenic activity as determined by in vitro E-screen assay and in vivo uterotrophic assay.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.1
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• pp.16-21
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• 2004

To assess the estrogenicities of Korean arrowroot (Pueraria thunbergiana) the contents of nine phytochemicals which are known to present were analyzed by HPLC. Also the estrogenecities of these phytochemicals were assayed using estrogen receptor dependent transcriptional expression assay. Daidzein and puerarin are major compounds in arrowroot, which were contained in the order of: root>stem>leaf>flower>seed. Among the assayed phytochemicals daidzein, genistein, biochanin A, formononetin, puerarin and genistin were highly estrogenic. The total estrogenicities in different parts of arrowroot and those by regions in Korea were also assessed. Roots had the highest estrogenic effects. The estrogenicities also were shown in stem and leaf. The differences in the estrogenicities of arrowroots by regions in Korea were shown. These results demonstrated that the estrogenic phytochemicals content was the highest in Korean arrowroot and also present in stem and leaf.