Salivary Cortisol and DHEA Levels in the Korean Population: Age-Related Differences, Diurnal Rhythm, and Correlations with Serum Levels

안련섭,이영진,최준영,권혁방,전세일 연세대학교의과대학 2007 Yonsei medical journal Vol.48 No.3

번식기에 개구리 여포의 스테로이드 생성변화와 난자의 성숙능 획득

안련섭,이용탁 한국통합생물학회 1994 동물학회지 Vol.37 No.2

3 종의 한국산개구리를 사용하여 여포들이 생성하는 스테로이드와 난자의 성숙능이 번식기 근처에서 어떻게 변하는가를 조사하였다. 개구리의 여포를 생체외 배양하면서 이들이 뇌하수체추출물(frog pituitary homogenate, FPH) 혹은 CAMP의 자극으로 생성하는 progesterone(Pa)과 testosterone(T)의 양을 측정하여 이로부터 P4yT의 비율을 구하였으며 여포난자들이 FPH. P4 혹은 12-0-tetrsdecanoyl phorbol-13-acetate(TPA)에 반응하여 성숙을 일으키는 시기를 조사하였다. 북방산개구리의 여포난자는 11월 부터 호르몬에 반응하여 성숙을 일으켰으며 2월에 PJT의 비율이 급격히 증가하였다 참개구리는 5월에 가서of PJT의 비율이 현저히 증가하고 FPH에 반응하여 성숙을 일으켰다. 그러나 P4에 의한 난자성숙은 3월부터 시작하였다 옴개구리는 7월에 P4/T 비율의 증가가 있었으며 이때부터 또한 TPA에 반응하여 난자의 성숙을 일으켰다 따라서 3종의 개구리에서 모두 각각의 번식기 근처에서 PJT의 비가 급격히 증가하고 또한 성숙능이 생긴다는 것을 알 수있었다.

북방산개구리 난자의 자발적 성숙에 관한 연구

안련섭,최한호권혁방배동규 한국통합생물학회 1993 동물학회지 Vol.36 No.1

본연구는 북방산개구리 여포난자의 자발적 성숙과 관련된 몇가지 요인들에 대해 조사하였다. 자발적 성숙을 일으키는 여포에서 여포조직을 제거하여 denuded 난자를 만든다음 배양했을 때에는 자발적 성숙을 일으키지 않았다. 이들 denuded 난자에 frog pituitary homogenate(FPH, 0.05 gland/ml)를 처리하여도 역시 성숙이 일어나지 않았으나 progesterone(P4, 0.5 Ig/ml)을 처리하였을 때에는 성숙이 일어났다. 동면기간에 여포들이 생성하는 P4와 testosterone(T)의 양을 측정해본 결과 동면초에는 비슷한 양의 P4와 T를 생성하였으나 후기에는 P4만 생성하고 T는 거의 생성하지 않았다. 배양액에 estradiol을 첨가하면 농도에 의존하여 FPH에 의해 촉진된 P4의 생성을 억제할 뿐 아니라 성숙도 비가역적으로 억제하였다. 배양액에 첨가한 hvpoxanthine은 CAMP와 항진적으로 난자의 성숙을 억제하였다. 이러한 결과들은 자발적 성숙과정에 여포세포와 P4/T 비율의 증가가 중요한 요인이 된다는 것과 일반적인 난자성숙억제제는 자발적 성숙을 역시 억제한다는 것을 보여준다.

Effect of 6-Hydroxydopamine (6-OHDA) on the Expression of Testicular Steroidogenic Genes in Adult Rats

허현진,안련섭,이성호 한국발생생물학회 2010 발생과 생식 Vol.14 No.3

A neurotoxin, 6-hydroxydopamine (6-OHDA) has been widely used to create animal model for Parkinson's disease (PD). The present study was undertaken to examine whether depletion of brain dopamine (DA) stores with 6-OHDA can make alteration in the activities of the testicular steroidogenesis in adult rats. Young adult male rats (3 months old) were received a single dose of 6-OHDA (200 ㎍ in 10 ㎕/animal) by intracerebroventricular (icv) injection, and sacrificed after two weeks. The mRNA levels of steroidogenesis-related enzymes were measured by qRT-PCRs. Serum testosterone levels were measured by radioimmunoassay. Single icv infusion of 6-OHDA significantly decreased the mRNA levels of CYP11A1 (control:6-OHDA group=1:0.68±0.14 AU, p<0.05), CYP17 (control:6-OHDA group=1:0.72±0.13 AU, p<0.05). There were no changes in the mRNA levels of 3β-HSD (control:6-OHDA group=1:0.84±0.08 AU) and 17β-HSD (control: 6-OHDA group=1:0.63±0.20 AU), though the levels tended to be decreased in the 6-OHDA treated group. Administration of 6-OHDA decreased significantly the mRNA level of StAR when compared to the level of saline-injected control animals (control:6-OHDA group=1:0.72±0.08 AU, p<0.05). Treatment with single dose of 6-OHDA remarkably lowered serum testosterone levels compared to the levels of control group (control:6-OHDA group=0.72±0.24:0.13±0.03 ng/㎖, p<0.05). Taken together with our previous study, the present study demonstrated that the activities of hypothalamus-pituitary-testis hormonal axis could be negatively affected by blockade of brain DA biosynthesis, and suggested the reduced reproductive potential might be resulted in the animals. More precise information on the testicular steroidogenic activities in PD patients and PD-like animals should be required prior to the generalization of the sex steroid hormone therapy to meet the highest standards for safety and efficacy.

수컷 흰쥐의 시상하부-뇌하수체 축 호르몬 유전자 발현에 미치는 6-Hydroxydopamine(6-OHDA)의 영향

허현진,안련섭,이성호 한국발생생물학회 2009 발생과 생식 Vol.13 No.4

6-hydroxydopamine(6-OHDA)는 파킨슨 질환 동물 모델의 제조에 널리 사용되는 신경독소로 도파민성 뉴런에 대한 특이적인 독성을 나타낸다. 도파민 신호는 중추신경계의 광범위한 영역에서 생리 기능을 조절하는데, 이에 따라 파킨슨병 환자와 6-OHDA를 처리한 동물들의 신경내분비 활성에 극심한 변화가 있을 것으로 예상할 수 있다. 하지만 6-OHDA 주사 모델에서 시상하부-뇌하수체 신경내분비 회로에 관한 연구들은 전무한 실정이다. 본 연구는 6-OHDA에 의한 뇌 카테콜아민 합성의 차단이 성체 수컷 흰쥐의 시상하부-뇌하수체 호르몬 유전자들의 전사 활성에 일으키는 변화를 조사한 것이다. 생후 3개월의 수컷 흰쥐(SD strain)에 개체 당 200 ㎍의 6-OHDA를 10 ㎕의 생리식염수에 녹여 뇌실 내 주사(icv)하였고, 2주 후에 모든 실험동물들을 희생시켰다. 시상하부-뇌하수체 호르몬 유전자들의 mRNA 수준을 조사하기 위해 total RNA를 추출하여 반-정량적 RT-PCR을 시행하였다. 카테콜아민 생합성에서 속도조절효소로 작용하는 tyrosine hydroxylase(TH)의 경우 6-OHDA군에서 대조군에 비해 유의한 발현 감소가 나타났고(대조군:6-OHDA군=1:0.72±0.02AU, p<0.001), 이를 통해 6-OHDA 주사의 효력을 확인하였다. 시상하부에서 gonadotropin-releasing hormone(GnRH)과 corticotropin releasing hormone(CRH)의 mRNA 수준은 6- OHDA군이 대조군에 비해 유의하게 낮았다(GnRH, 대조군:6-OHDA군=1:0.39±0.03AU, p<0.001; CRH, 대조군:6-OHDA군=1:0.76±0.07AU, p<0.01). 뇌하수체에서 glycoprotein hormone들의 공통적인 alpha subunit(Cgα)과 LH beta subunit(LH-β) 그리고 FSH beta subunit(FSH-β)의 mRNA 수준의 경우 모두 6-OHDA군에서 대조군에 비해 유의한 감소를 나타냈다(Cgα, 대조군:6-OHDA군=1:0.81±0.02AU, p<0.001; LH-β, 대조군:6-OHDA군=1:0.68±0.04AU, p<0.001; FSH-β, 대조군:6-OHDA군=1:0.84±0.05AU, p<0.01). 이와 유사하게, 6-OHDA군에서의 뇌하수체 adrenocorticotrophic hormone(ACTH) 전사 수준 역시 대조군에 비해 유의하게 낮았다(대조군:6-OHDA군=1:0.86±0.04AU, p<0.01). 본 연구는 중추신경계로의 도파민 신경독소 주입에 의해 두 가지의 시상하부-뇌하수체 신경내분비 회로인 GnRH-성선자극호르몬 회로와 CRH-ACTH 회로의 전사 활성이 하향조정됨을 증명하였다. 이러한 결과는 시상하부로의 CA입력은 시상하부-뇌하수체 기능 조절을 통해 생식소와 부신의 활성에 영향을 미침을 시사하는 것으로, 파킨슨병 환자들에게서 빈번하게 발생하는 성 기능 장애와 열악한 스트레스 반응을 설명할 단서를 제공한다.

Effects of Azoles on the In vitro Follicular Steroidogenesis in Amphibians

김안나,안련섭,권혁방 한국통합생물학회 2006 Animal cells and systems Vol.10 No.4

Azoles are widely used antifungal agents, whichinhibit the biosynthesis of fungal cell-membrane ergosterol.In this study, using an amphibian follicle culture system, theeffects of azoles on follicular steroidogenesis in frogs wereexamined. Itraconazole (ICZ), clotrimazole (CTZ) andketoconazole (KCZ) suppressed pregnenolone (P5) productionby the follicles (ED50; 0.04M, 0.33M, and 0.91M,(FPH). However, fluconazole (FCZ), miconazole (MCZ) andeconazole (ECZ) were not effective in the suppression of P5production. Not all the azoles examined suppressed theconversion of exogenous P 5 to progesterone (P4) (by 3β-HSD) or P4 to 17α-hydroxyprogesterone (17α-OHP) (by17α-hydroxylase), or androstenedione (AD) to testosterone(T) (by 17β-HSD). In contrast, CTZ, MCZ and ECZ inα-OHP toAD (by C17-20 lyase) (ED50; 0.25M, 4.5M, and 0.7M,respectively) and CTZ, KCZ, ECZ and MCZ stronglysuppressed the conversion of exogenous T to estradiol (E2)(by aromatase) (ED50; 0.02 M, 8M, 0.07M, 0.8M,respectively). These results demonstrated that some azoleand particularly, P450scc and aromatase are more sensitiveto azoles than other steroidogenic enzymes.

Age-related Changes in Luteinizing Hormone and Testosterone Levels in Korean Men

이성호,안련섭,권혁방 한국발생생물학회 2008 발생과 생식 Vol.12 No.1

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, ; 30s, ; and 40s, ), 50대 이상에서 유의하게 증가하였다(50s, and 60s, ). 또한, 혈중 T 수준도 40대까지는 변화하지 않았으나(20s, , 30s, , 40s, ng/mL), 50대 이후 유의하게 감소하였다( Changes in luteinizing hormone (LH), serum testosterone (T), and salivary T levels with age were examined in Korean men. Serum was obtained from 167 Korean men of different ages (), and the serum LH and T levels were measured. Saliva samples were also obtained, and the salivary T level was determined. The LH levels did not change considerably until 40 y of age (20s, ; 30s, ; and 40s, ) but increased significantly around 50 y (50s, and 60s, ). Further, the serum T levels also did not change until 40 y of age (20s, , 30s, , 40s, ) but decreased significantly at 50 y (50s, ; 60s, ). The salivary T levels also showed small changes until the age of 40 y (, ) but decreased significantly at 50 y (). Thus, the relative ratio of salivary T to serum T levels did not change significantly in all the ages examined (). Linear regression analysis predicted that the LH levels increased 1.5%/y while the serum and salivary T levels decreased 1%/y and 0.8%/y, respectively. The serum T/LH ratio did not change considerably until the age of 40 y (, ) but decreased significantly () at 50 y. Age-related changes in the salivary T/LH ratio were very similar to those in the serum T/LH ratio. These results demonstrated that LH and T levels in serum or saliva did not change considerably until 40 y of age; instead, in Korean men, from 50 y of age, the LH level increased, while the T level decreased. This suggests that primary testicular failure that occurred due to aging (approximately 50 y) and caused this phenomenon. The present study also shows that the salivary T level can be an indicator of the free T level in serum although the salivary T level correlates weakly with the total T level in serum (r=0.53). Thus, information regarding salivary T levels may be useful for studying the age-related changes occurring in male testicular physiology.

Cortisol Awakening Response and Nighttime Salivary Cortisol Levels in Healthy Working Korean Subjects

신일영,안련섭,전세일,이영진,김민수,이채관,성시몬 연세대학교의과대학 2011 Yonsei medical journal Vol.52 No.3

Cortisol awakening response (CAR) and nighttime cortisol levels have been used as indices of adrenocortical activity. However, population-based statistical information regarding these indices has not been provided in healthy subjects. This study was carried out to provide basic statistical information regarding these indices. Materials and Methods: Cortisol levels were measured in saliva samples collected immediately upon awakening (0 min), 30 min after awakening and in the nighttime on two consecutive days in 133 healthy subjects. Results: We determined the mean [standard deviation (SD)], median (interquartile range) and 5th-95th percentile range for each measure and auxiliary indices for CAR, i.e., the secreted cortisol concentration within 30 min of awakening (CARscc) and absolute and relative increases in cortisol level within 30 min of awakening (CARi and CARi%, respectively). We also determined these values for auxiliary indices derived from nighttime cortisol level, i.e., the ratio of cortisol level 30 min after awakening (CA_30 min) to nighttime level (CA_30 min/NC), as well as absolute and relative decreases in cortisol levels from CA_30 min to nighttime (DCd and DCd%, respectively). We found no significant differences in cortisol level for any time point or in auxiliary indices between collection days, genders and ages. Conclusion: The provided descriptive information and statistics on the CAR and nighttime cortisol level will be helpful to medical specialists and researchers involved in hypothalamus-pituitary-adrenal axis assessment.

Further Evidences For the Two-cell Type Model in the ovarian Follicular Steroidogenesis in Amphibians

유명식,안련섭,강해묵,권혁방 한국통합생물학회 1995 한국동물학회 뉴스레터 Vol.12 No.2

Effects of Heavy Metals on the in vitro Follicular Steroidogenesis in Amphibians

최미정,안련섭,권혁방 한국통합생물학회 2006 Animal cells and systems Vol.10 No.4

Heavy metals are well known as importantenvironmental pollutants and also considered as endocrinedisrupters. This study was performed to evaluate the directeffects of heavy metals such as cadmium (Cd), zinc (Zn),mercury (Hg), lead (Pb), cobalt (Co), and arsenic (As) on thevarious steroidogenic enzymes in frog ovarian follicles.Ovarian follicles from Rana catesbeiana were isolated andhomogenate (FPH, 0.05 gland/ml) or various steroid precursorswith or without heavy metals (0.01-100M), and steroidlevels in the follicle or culture medium were measured byradioimmunoassay (RIA). Thus, the steroidogenic enzymeactivities were indirectly evaluated by measuring theconverted steroid levels from the added precursor steroid.Among heavy metals, Hg, Cd and Zn significantly inhibitedFPH-induced pregnenolone (P5) production by the follicles(EC50, 4.0M, 25.6M and 5.7 M, respectively ), and alsosuppressed the conversion of testosterone (T) to estradiol17β (E2) (EC50, 4.2M, 7.5M and 80.0 M) while Pb, Coand As are not or less effective in the inhibition. Otherenzymes such as C 17-20 lyase and 17 β-hydroxysteroiddehydrogenase (17 β-HSD) were suppressed only in thehigh concentration of Hg, Cd and Zn. Taken together, thesedata demonstrate that cytochrome P450 side chain cleavage(P450scc) and aromatase are much more sensitive to heavymetals than other steroidogenic enzymes and Hg, Cd andZn show stronger toxicity to follicles than other heavy metalsexamined.