• Clinical and Experimental Reproductive Medicine
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• 제36권4호
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• pp.265-274
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• 2009

목 적: 미세리보핵산 (microRNA, miR)은 전사 후 (post-transcriptional) 단계에서 목표 유전자 (target gene)의 발현을 억제하여 세포의 발달과 성장에 중요한 역할을 하는 것으로 알려져 있다. 그러나 난포의 성장과정 중의 miR 발현 양상에 대해서는 잘 알려져 있지 않다. 따라서 존 연구는 생쥐 난포의 체외배양 후 생식샘자극호르몬 (gonadotropin)과 사람융모성생식샘자극호르몬 (hCG) 첨가에 따른 난자와 난구세포에서의 miR 발현 양상을 살펴보고자 수행되었다. 연구방법: 생후 12일된 생쥐 (C57BL6)의 난소 적출 후, 전동 난포 (preantral follicle)를 분리하여 무작위로 $20\;{\mu}L$ 점적의 배양액만 있는 군 (control group), 재조합 난포자극호르몬을 첨가한 군 (FSH group), 재조합 황체형성호르몬을 첨가한 군 (LH group), FSH와 LH를 같이 첨가한 군 (FSH+LH group)으로 나누어 배양하였다. 난포가 충분히 성장하였을 때, 다시 hCG를 첨가한 군 (hCG (+) group)과 첨가하지 않은 군 (hCG (-) group)으로 나누어 hCG (-) group에서 난자와 난구세포를 각각 분리하여 RNA를 추출하였다. 36시간 후, 배란된 난자 난구세포 복합체 (cumulus oocyte complex, COC)에서 난자와 난구세포를 각각 분리하여 RNA를 추출하고, mmu--miR-16, -miR-27a, -miR-126, -miR-721 등의 miR에 대한 primer를 이용하여 실시간 중합연쇄반응을 시행하였다. 결 과: 배란율과 MII 난자 생성율은 다른 군들에 비해 FSH+LH군에서 유의하게 높았다. 각 군내의 난자와 난구세포 사이에서도 miR 발현 양상의 차이가 관찰되었다. 또한, 난자와 난구세포에서의 miR 발현 양상은 각 군간 차이가 있었으며, hCG (+)군과 hCG (-)군간에도 차이를 나타냈다. 결 론: 생쥐 난포의 체외배양 중 난자 및 난구세포에서의 miR 발현 양상은 gonadotropin의 종류 및 난자의 성숙도에 따라 다르다. 이러한 결과는 표적 유전자의 발현에 대한 추후 연구를 통한 확인이 필요할 것으로 사료된다.

• Asian-Australasian Journal of Animal Sciences
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• 제20권3호
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• pp.340-350
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• 2007

The objective of this study was to determine whether peroxisome proliferator-activated receptor ${\gamma}$(PPAR${\gamma}$ is involved in the regulation of weaning to estrus of primiparous sows. Twelve sows composed of 6 groups of 2 full-sibs in a similar age (325.2 d), body weight (BW; 152.4 kg) and backfat thickness (BFT; 27.0 mm) at start of lactation, were allocated to accept 31 MJ (restricted group, R-group) or 53 MJ (control group, C-group) DE/d treatment, respectively. The experimental results indicated that the low energy intake resulted in excessive losses of BW and BFT during lactation in R-group sows, which may be related to decrease of serum 15-deoxy-${\Delta}^{12,14}$-prostaglandin $J_2$ (15d-$PGJ_2$), a ligand of PPAR${\gamma}$ The obvious peak and the frequency of LH, FSH and estradiol ($E_2$) were only observed in C-group sows. Except for $E_2$ at d 1 and 2, serum FSH, LH and $E_2$ concentrations in R-group were lower than those in C-group sows after weaning. However, the serum progesterone ($P_4$) level in R-group sows was always more than that in C-group. The expression abundances of PPAR${\gamma}$and GnRH receptor (GnRH-R) in pituitary, FSH receptor (FSH-R), LH receptor (LH-R), estrogen receptor (ES-R) and aromatase in ovary of anestrous sows were lower than those of estrous sows. Neither the BFT nor the BW was associated with the mRNA abundance of PPAR${\gamma}$in hypothalamus during lactation. Expressions of PPAR${\gamma}$in pituitary and ovary were affected evidently by the BFT changes and only by the loss of BW of sows during and after lactation. Furthermore, PPAR${\gamma}$mRNA level in ovary was significantly related to the expression abundances of GnRH-R, FSH-R, ES-R and aromatase, and GnRH-R was obviously associated with PPAR${\gamma}$expression in pituitary. However, PPAR${\gamma}$expression in hypothalamus likely has no effects on these genes expression and no obvious difference for all sows. Not serum $E_2$ or $P_4$ alone but the ratios of $E_2$ to $P_4$ and 15d-$PGJ_2$ to $P_4$, and serum FSH and LH were evidently related to PPAR${\gamma}$expression in pituitary and ovary. It is concluded that PPAR${\gamma}$is associated with body conditions, reproduction hormones and their receptor expression, which affected the functions of pituitary and ovary and ultimately the estrus after weaning of primiparous sows.

• Journal of Acupuncture Research
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• 제17권1호
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• pp.287-318
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• 2000

The purpose of this study is to observe the effect of Aqua-acupuncture utilizing Lycopi Herba Solution(LH-AS) on arthritis. For that purpose, we formed three experimental group, synovial cells of human body, normal BALB/C mice and DBA/1J mice with collagen II-induced arthritis, and measured the treatment effect of LH-AS on each group. The conclusions are as follows. 1. After the LH-AS treatment on synovial cells, there were no significant change in 1, 10, 50 and $100{\mu}g/m{\ell}$ whereas there was significant change in 200 and $400{\mu}g/m{\ell}$ in cytotoxicity. 2. IL-6, IL-$1{\beta}$, TNF-${\alpha}$ gene expression of synovial cells and the secretion amount of IL-6 and IL-$1{\beta}$ are significantly inhibited in treatment group with LH-AS. 3. The proliferation of synovial cells was significantly inhibited in treatment group with rIL-6, LH-AS 200 and rIL-6, $100{\mu}g/m{\ell}$. 4. After the DBA/1J mice with collagen II-induced arthritis were treated by LH-AS, the incidence of arthritis, hind paw edema, the index of arthritis and DTH were significantly inhibited. 5. After the DBA/1J mice with collagen II-induced arthritis were treated by LH-AS, splenetic weight was significantly increased and the number of leukocyte was significantly decreased. But there was no significant change in the number of platelet. 6. After the DBA/1J mice with collagen II-induced arthritis were treated by LH-AS, the number of $CD4^+$, $CD8^+$ activated cells and the surface-receptor expression were significantly increased whereas the number of $CD19^+$ activated cells and the surface-receptor expression were decreased. 7. After the DBA/1J mice with collagen II-induced arthritis were treated by LH-AS, total protein, LDH were significantly decreased, but there was no significant change in creatinine. 8. After the normal splenetic cells of BALB/C mice were treated by LH-AS and cultured, it was observed that the adherent cells were morphologically activated and IL-12 and IFN-${\gamma}$ gene expression were increased. 9. After the normal splenetic cells of BALB/C mice were treated by LH-AS and cultured, the number of $CD4^+$, $CD8^+$, $CD19^+$ activated cells and surface-receptor expression were inhibited when being compared with the control group. Taking all these observations into account, LH-AS injection is considered to be effective in treating arthritis and put to practical use in future arthritis clinic.

• Clinical and Experimental Reproductive Medicine
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• 제29권1호
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• pp.67-76
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• 2002

Objective: To assess the difference of baseline hormonal status and pathophysio logy, and confirm the risk factors for long term complication according to Body Mass Index in women with polycystic ovary syndrome. Materials and Methods: Serum level of LH, FSH, Estradiol, Prolactin, Testosterone, DHEA-S, fasting insulin were measured and 100 gm oral glucose tolerance test and endometrial biopsy were performed in total 75 chronic anovulation patients and 20 normal cycling infertility patients. 95 evaluated patients were divided into 3 groups including patients with chronic anovulation having BMI below 25, BMI beyond 25.1, normal cycling infertility patients, Group 1 (n=39), Group 2 (n=36), Group 3 (n=20), respectively. Statistical analysis was performed respect to relationship between BMI and measured hormone level, sum of glucose level during 100 gm OGTT, insulin resistance using t-test, ANOVA test, Post Hoc test, Mann-Whitney test. p<0.05 was considered as statistically significant. Results: Serum LH level and LH/FSH ratio was significantly higher in Group 1, compared than Group 2 or 3 (p<0.05), BMI and LH, LH/FSH ratio was negatively correlated (r=-0.351, r=-0.318). There was no significant difference according to BMI in FSH, testosterone, estradiol, prolactin, DHEA-S level. Fasting insulin and sum of glucose level during 100 gm OGTT were significantly higher in Group 2 compared than Group 1 or Group 3 (p<0.05), there was no significant difference between Group 1 and Group 3. Insulin resistance was more frequently identified in Group 2 compared than Group 1 (p=0.001). Conclusions: BMI and LH, LH/FSH ratio were negatively correlated, so clinical significance of LH, LH/FSH ratio in diagnosis of PCOS may be attenuated by increasing body weight. Overweight patients with chronic anovulation may be the risk group for developing insulin resistance, hyperinsulinemia, glucose intolerance, later type 2 DM. Hyperinsulinemia may operate mainly in overweight chronic anovulation patients in development of hyperandrogenism.

• 대한임상검사과학회지
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• 제37권3호
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• pp.143-148
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• 2005

Automatic hematology analyzers provide the leukocyte differential count and the useful flags of the various hematological parameters. We compared LH 750 (Beckman Coulter Co., Miami, FL, USA) with manual method on differential leukocyte counts and evaluated the usefulness of the suspect flags, provided by this instrument. The comparison of leukocyte differential counts between two methods showed good correlation coefficient (r), which were 0.95 (neutrophil), 0.92 (lymphocyte), 0.82 (monocyte) and 0.95 (eosinophil). The frequency of the total flags displayed on LH 750 were 15.5%, which included immature granulocyte/left shift 63.5%, nucleated RBC 14.6%, platelet clumps 13.1%, variant lymphocyte 50% and blast 16.6%. This instrument showed higher positive predictive value in the flags such as platelet clumps 68.8%, immature granulocyte/left shift 61.5%, nucleated RBC 27.3%, variant lymphocyte 50% and blast 16.7%. In this study, the leukocyte differential counts of LH 750 showed good correlation with manual method and the suspect flags also showed a good performance for applying the criteria of re-examination in the clinical laboratory.

• 한국발생생물학회지:발생과생식
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• 제12권1호
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• pp.57-66
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• 2008

한국 남성의 연령 증가에 따른 혈중 luteinizing hormone(LH), testosterone(T), 그리고 타액 T 수준 변화를 조사하였다. 혈중LH 수준은 40대까지 유의한 변화를 보이지 않았으나(20s, $2.5{\sim}1.0$; 30s, $2.7{\sim}1.5$; and 40s, $2.5{\sim}1.8\;mIU/mL$), 50대 이상에서 유의하게 증가하였다(50s, $3.7{\sim}1.8$ and 60s, $3.1{\sim}1.7\;mIU/mL$). 또한, 혈중 T 수준도 40대까지는 변화하지 않았으나(20s, $5.3{\sim}2.6$, 30s, $4.4{\sim}1.4$, 40s, $4.1{\sim}1.5$ ng/mL), 50대 이후 유의하게 감소하였다(50s, $3.4{\sim}1.5$; 60s, $2.6{\sim}0.8\;ng/mL$). 타액 T 수준 또한 40대까지 약간의 변화가 나타났으나($20s{\sim}40s$, $0.11{\pm}0.015\;ng/mL$), 50대에 유의하게 감소하였다($0.08{\sim}0.03\;ng/mL$). 타액 T 대 혈중 T의 상대적인 비율은 모든 연령대에서 유의한 변화가 없었다($2.4{\sim}0.9%$). 직선회귀 분석(Linear regression analysis)에서 혈중 LH 수준은 매년 1.5%씩 증가하고 혈중 T와 타액 T 수준은 각각 매년 1%와 0.8%씩 감소하는 것으로 예측되었다. 혈중 T/LH 비율은 40대까지 유의한 변화가 없었으나($20s{\sim}40s$; $2.27{\pm}0.14$) 50대에 유의하게 감소하였다($1.2{\sim}1.0$). 연령과 관련된 타액 T/LH 비율은 혈중 T/LH 비율과 대단히 유사하였다. 본 연구 결과에서 혈중 LH와 T, 그리고 타액 T 수준이 한국인 남성에서 40대까지는 유의하게 변화하지 않았고, 50대부터는 LH 수준이 증가하고 T 수준이 감소하였다. 이 결과는 50대경 노화과정에 의해 일어나는 원발성 정소부전(primary testicular failure)이 원인인 것으로 보인다. 또한, 본 연구결과는 비록 타액 T 수준이 혈중 총 T 수준과 약한 상관관계(r=0.53)를 갖지만, 혈중 free T 수준을 나타내는 것으로 보인다. 그러므로, 타액 T 수준에 대한 정보는 연령과 관련해서 나타나는 남성 정소생리를 연구하는데 유용할 것으로 사료된다.

• 환경생물
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• 제21권3호
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• pp.308-312
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• 2003

Various pesticides known or suspected to interfere with steroid hormone function were screened toy effects in leydig cells on catalytic activity and mRNA expression of aromatase. Dichlorodiphenyltrichloroethane (DDT) is a widespread environmental pollutant. In this study, we investigated the effect of DDT on testosterone production through aromatase activity and its molecular mechanism in testicular leydig cell, R2C by using radioimmunoassay (RIA). As the results, the potent leydig: cell activator LH increased testosterone production compared to the control. DDT exposure significantly decreased testosterone production in R2C cell. In addition, DDT was found to increase aromatase gene expression and activity in R2C cell in a dose dependent manner. In order to assess whether the suppressive effects of DDT on LH-inducible testosterone (T) production might be influenced by the ER, ICI 182.780 was used, and it was found that these inhibitory effects of DDT were antagonized by ICI 182.780, implying that the estrogen receptor (ER) mediates the suppressive effects of DDT. Furthermore, the inducible effects of DDT on aromatase gene expression might be influenced by the ER, ICI 182.780 was used, and it was found that these enhancing effects of DDT were antagonized by ICI 182.780, implying that the ER mediates the inducible effects of DDT. Our results indicated that DDT inhibition of luteinizing hormone (LH) -inducible T production in R2C cell is mediated through aromatase. However, the precise mechanisms by which DDT enhance in R2C cell remains unknown. The current study suggests the possibility that DDT might act as a modulator aromatase gene transcription.

• Asian-Australasian Journal of Animal Sciences
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• 제11권1호
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• pp.78-83
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• 1998

The objective of this study was to investigate the responsiveness of hypophysis and gonads to synthetic GnRH among prepubertal buffalo heifers at 12 months of age. Peripheral plasma FSH, LH, estradiol and progesterone level were measured in blood samples collected at 1 hr before and up to 18 days subsequent to the administration of $200{\mu}g$ GnRH (n=6) or saline (n=6) in Murrah buffalo heifers. The pretreatment peripheral plasma FSH, LH, estradiol and progesterone among GnRH treated heifers were $7.35{\pm}0.45ng/ml$, $1.08{\pm}0.3ng/ml$, $22.93{\pm}1.06pg/ml$ and $0.27{\pm}0.04ng/ml$ respectively. A quick elevation (p < 0.01) of FSH and LH within five min of GnRH administration was observed in all geifers. Although the peak FSH $(89.57{\pm}23.43ng/ml)$ and LH $(7.52{\pm}3.08ng/ml)$ reached by 10 min of GnRH administration, yet the animals differed both in terms of their amplitude response of FSH and LH release as well as in terms of time which animals took to exhiit maximum response to GnRH administration. The GnRH administration did not cause alteration in plasma estradiol and progesterone level. The present study suggests that the pituitary of 12 month buffalo heifers has capacity to synthesize and store of gonadotropin and have developed receptors for GnRH for a spike of gonadotropin release.

• 한국발생생물학회지:발생과생식
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• 제11권1호
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• pp.49-54
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• 2007

Ethane 1,2-dimethane sulfonate(EDS)은 Leydig cells(LC)만을 선별적 사멸을 유도하는 약물로서 가역적인 테스토스테론(testosterone, T) 결핍 흰쥐 모델을 만드는데 널리 사용된다. 본 연구에서는 수컷 흰쥐 뇌하수체의 생식소자극호르몬인 LH와 FSH의 발현에 미치는 EDS 투여 효과를 조사하였다. 성숙한 수컷 흰쥐(SD strain, $300{\sim}350\;g$ B.W.)에 EDS(75 mg/kg, i.p.)를 1회 복강주사하고 주사 후 0, 1, 2, 3, 4, 5, 6 그리고 7주가 경과한 날 희생시켰다. 뇌하수체로부터 total RNA를 추출한 후 뇌하수체 glycoprotein hormone common alpha subunit($C{\alpha}$), LH beta subunit($LH{\beta}$), FSH beta subunit($FSH{\beta}$) 그리고 GnRH 수용체(GnRH-R)의 발현 변화를 semi-quantitative RT-PCR로 측정하였다. 그 결과, $C{\alpha}$ 전사수준은 주사 후 1주부터 급격히 상승하여 주사 후 4주까지 유의하게 높게 유지되다가 5주 후부터 control 수준으로 회귀하였다. $LH{\beta}$ 전사 수준은 주사 후 2주부터 유의하게 상승하여 주사 후 4주에 최고 수준에 도달하였으며, 5주 후부터 control 수준으로 감소하였다. $FSH{\beta}$ 전사수준은 주사 후 2주부터 유의하게 상승하여 주사 후 3주에 최고 수준에 도달하였으며, 4주 후부터 감소하여 5주 후에 최소치를 보였다. 유사하게, GnRH-R 전사 수준도 주사 후 2주부터 유의하게 상승하여 주사 후 3주에 최고 수준에 도달하였으며, 5주 후부터 control 수준으로 감소하였다. 본 연구는 EDS 주사에 의해 수컷 흰쥐 뇌하수체 전엽에서의 생식소 자극호르몬 subunit들과 GnRH-R의 발현 변화가 가역적으로 유도될 수 있음을 보여준 것이다. EDS 주사 모델은 수컷 흰쥐에서의 시상하부-뇌하수체 신경내분비 축의 호르몬 조절에 대한 기작을 이해하는데 도움이 될 것이다.

• Environmental Analysis Health and Toxicology
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• 제18권2호
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• pp.95-100
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• 2003

본 연구는 환경호르몬(endocrine disruptors)으로 분류되었으며, 제초제로 널리 사용되었던 Dichlorodiphenyltrichloroethane(DDT)가 설치류의 생식세포 중 Leydig 세포의 testosterone(T)생성억제 및 그 관련 작용메카니즘을 규명코자 수행되었다. 먼저 흰쥐의 웅성 생식세포주인 R2C세포에 T의 양 및 aromatase 활성도를 radio immunoassay (RIA)방법을 이용하여 측정하였다. 그 결과 R2C 세포에 황체형성호르몬(LH)를 처리하여 testosterone생성을 증가시킨 후, DDT를 처리한 군들은 대조군에 비하여 농도 의존적으로 T의 양은 감소하였으며, aromatase 활성도는 증가하였다. 또한 DDT자체만 처리한 군에서도 대조군에 비하여 testosterone의 생성이 감소하였다. 이러한 aromatase활성 증가가 estradiol receptor(ER)와의 상호 관련성을 확인하기 위해 ER antagonist인 ICI 182.780를 처리한 후 T의 양 및 aromatase 활성도를 측정한 결과 DDT에 의해 증가된 aromatase활성도가 ICI 182.780에 의해 다시 감소됨을 확인하였다. 또한 DDT에 의해 감소된 T의 양도 ICI 182.780에 의해 다시 회복되었다. In vivo실험으로 흰쥐에 DDT를 직접 투여한 후 정소 내 성 호르몬들을 측정해 본 결과 T의 양은 유의성 있게 감소하였으며, estradiol(E$_2$)의 양은 증가하였으며, aromatase 활성도도 감소하였다. 이러한 결과를 종합해 볼 때 DDT는 aromatase를 감소시키고, 이렇게 감소된 aromatase에 의해 testosterone 생성량을 억제하고, 이러한 DDT의 aromatase의 감소는 ER을 경유하는 것으로 추정할 수 있다.