• 분석과학
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• 제24권2호
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• pp.142-149
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• 2011

Resveratrol을 GC/TOF-MS를 이용하여 대사체를 확인한 결과, 두 개의 phenyl기를 연결하는 이중결합이 단일결합으로 환원된 구조로 추정되었다. 또한, GC/MSD를 이용하여 resveratrol 및 내인성스테로이드의 분석법에 대한 유효성을 점검한 결과, 회수율은 96.47 - 114.74%의 범위로 나타났으며, intra-day와 inter-day의 정밀도는 1.40 - 10.87%과 1.10 - 10.93% 그리고 정확도는 80.03 - 119.92%과 80.02 - 119.56%로 조사되었고, 모두 0.996이상의 직선성을 나타내어 유효한 분석 방법으로 검증되었다. 한편, 지원자들에게 resveratrol을 경구투여 한 요시료로부터 resveratrol과 그 대사체에 대한 상관성을 조사해본 결과, 요중 최대농도 도달시간이 일반적인 약물(1 - 2시간) 보다 긴 10 - 15시간에 나타났으며, 대사체로의 전환율은 남성보다 여성이 높게 나타났다. 한편, 내인성 스테로이드는 약물 복용 후 20시간 까지는 resveratrol 및 그 대사체와 다소 유사한 분비형태를 나타내었으며, estrone과 estradiol의 경우 여성이 남성에 비해 이 약물에 대한 민감성이 높게 나타났다. 그 외의 내인성 스테로이드는 유의할만한 차별된 분비형태의 변화가 나타나지 않았다. 따라서 resveratrol의 경우 약물의 활성이 남성보다는 여성에게 유의한 영향을 미치는 것으로 추측되었다.

• Mass Spectrometry Letters
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• 제2권2호
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• pp.33-36
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• 2011

The misuse of anabolic androgenic steroids is of particular concern in sports and society. Thus, it is of great importance to discriminate endogenous steroids such as testosterone or testosterone prohormones from their chemically identical synthetic copies. In this study, gas chromatography-combustion/isotope ratio mass spectrometric (GC-C/IRMS) method has been developed and validated for discriminating the origin of anabolic androgenic steroids. The method involves the solid-phase extraction, enzymatic hydrolysis with ${\beta}$-glucuronidase, HPLC-fractionation for the cleanup and analysis by GC-C/IRMS. The difference(${\Delta}^{13}C$) of urinary ${\delta}^{13}C$ values between synthetic analogues and endogenous reference compounds (ERC) by GC-C/IRMS was used to elucidate the origin of steroids, and intra- and inter-day precision, specificity and isotope fractionation were evaluated. The present GC-C/IRMS method combined with HPLC cleanup was accurate and reproducible enough to be successfully applied to the test of urine sample from suspected anabolic steroid abusers.

• 대한화학회지
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• 제41권8호
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• pp.406-413
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• 1997

현재 시행되고 있는 GC/MS를 이용한 금지 약물 분석법을 이용하여 신체의 성장, 분화 및 대사에 있어서 중요한 역할을 하고 있으며, 근육 강화제의 복용이 체내 농도에 영향을 미치는 것으로 알려진 내인성 스테로이드의 동시 분석 조건을 설정하였다. 그 결과 72.33%~94.54%의 회수율을 얻었으며, within-a day 및 day-to-day 분석에서의 RSD 값은 각각 1.43%∼10.86%, 0.96%∼9.98%로 측정되었다. 이 방법을 이용하여 9가지 근육 강화제를 정상인에게 투여한 후, 시간에 따라 소변으로 배설되는 근육 강화제의 농도와 8종의 주요 내인성 스테로이드의 농도를 동시에 정량함으로서, 한국인을 기준으로한 근육 강화제의 농도 변화에 따른 내인성 스테로이드의 농도 및 그들간의 농도 비율의 변화 경향을 조사하였다.

• 분석과학
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• 제11권5호
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• pp.353-359
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• 1998

국제 올림픽 위원회에서 금지하고 있는 합성 동화성 스테로이드로서 다양한 생물학적 효과를 나타내고 있는 danazol을 대상으로 androgenic 효과를 확인하기 위해 성인 남자 3명에게 100 mg씩 복용하게 한 후 그들의 소변을 채취하였다. 인체내 대사과정에서 생성되는 주요 대사체인 ethisterone의 배설에 따른 8가지의 내인성 스테로이드들의 배설정도를 GC/MS의 selected ion monitoring(SIM) 방법으로 동시분석 하였다. 그 결과 본실험방법에서 내인성 스테로이드들에 대한 회수율은 71.59%~93.56% 이었으며, within-a day 및 day-to-day 분석에서의 RSD 값은 각각 1.87%~10.48%와 1.32~11.25%이었다. 이때의 검출한계는 $0.01{\sim}0.05{\mu}g/mL$ 이었다. 각각의 내인성 스테로이드들과 ethisterone의 표준물질을 뇨시료에 0.1, 0.5, 1.0, 10, 20 그리고 $50{\mu}g/mL$를 첨가하여 작성한 검정곡선은 전체적으로 상관계수 0.963~0.991의 직선성을 보였다. 따라서 본 실험은 내인성 스테로이드들의 변화를 관찰함으로서 간접적으로 스테로이드 계열의 금지약물 복용여부를 판단할 수 있는 방법으로 매우 유용함을 알 수 있었다.

• 대한약학회:학술대회논문집
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• 대한약학회 2001년도 Proceedings of the Pharmaceutical Society of Korea
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• pp.133.2-134
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• 2001

• The Korean Journal of Physiology
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• 제21권1호
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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.

• Journal of Microbiology and Biotechnology
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• 제31권3호
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• pp.464-474
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• 2021

Bacterial cytochrome P450 (CYP) enzymes are responsible for the hydroxylation of diverse endogenous substances with a heme molecule used as a cofactor. This study characterized two CYP154C3 proteins from Streptomyces sp. W2061 (CYP154C3-1) and Streptomyces sp. KCCM40643 (CYP154C3-2). The enzymatic activity assays of both CYPs conducted using heterologous redox partners' putidaredoxin and putidaredoxin reductase showed substrate flexibility with different steroids and exhibited interesting product formation patterns. The enzymatic characterization revealed good activity over a pH range of 7.0 to 7.8 and the optimal temperature range for activity was 30 to 37℃. The major product was the C16-hydroxylated product and the kinetic profiles and patterns of the generated hydroxylated products differed between the two enzymes. Both enzymes showed a higher affinity toward progesterone, with CYP154C3-1 demonstrating slightly higher activity than CYP154C3-2 for most of the substrates. Oxidizing agents (diacetoxyiodo) benzene (PIDA) and hydrogen peroxide (H2O2) were also utilized to actively support the redox reactions, with optimum conversion achieved at concentrations of 3 mM and 65 mM, respectively. The oxidizing agents affected the product distribution, influencing the type and selectivity of the CYP-catalyzed reaction. Additionally, CYP154C3s also catalyzed the C-C bond cleavage of steroids. Therefore, CYP154C3s may be a good candidate for the production of modified steroids for various biological uses.

• Clinical and Experimental Reproductive Medicine
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• 제28권4호
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• pp.279-286
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• 2001

Objective: To elucidate 1) whether there are any differences in the urine concentrations of steroid hormone metabolites between patients with leiomyoma and normal controls 2) the correlation between urinary profiles of steroid hormones and leiomyomas of the uterus according to their type, location, volume, and weight. Materials of Methods : The study population consisted of 37 premenopausal patients with uterine leiomyoma and the control group consisted of 25 premenopausal normal volunteer women without uterine leiomyoma. Confirmation of the existence of uterine leiomyoma was done by ultrasonography and histopathological examination after surgery. The volume of the leiomyoma was estimated by trans-abdominal and/or trans-vaginal ultrasonography. The Leiomyomas were divided into 3 types (subserosal, intramural and submucosal). Seventeen patients had subserosal type of leiomyoma, 10 with the intramural type and 10 with the submucosal type. The locations of the leiomyoma were also divided into 3 groups (fundus, body and isthmus). Seventeen patients showed a fundus location, 10 in body, and 10 in isthmus. We compared urinary profiles of the endogenous steroids between patients with leiomyomas and normal controls, and also investigated the relationship between urinary profiles of the endogenous steroids and leiomyomas according to their type, location, volume and weight by using highly sensitive Gas Chromatography-Mass Spectrometry (GC-MS) system. Results: The mean ages of the patients with leiomyomas and the control group were $43.1{\pm}5.6$ and $40.6{\pm}7.2$ years, the weights were $63.4{\pm}7.3$ and $59.4{\pm}8.1\;kg$, and their heights were $155.4{\pm}4.8$ and $159.3{\pm}4.8\;cm$ respectively. Seventeen patients had subserosal, 10 had intramural, and 10 had submucosal leiomyomas. There were 17 patients with leiomyoma located in fundus, 10 in body and 10 in isthmus. $17{\beta}$-estradiol, 5-AT, 11-keto ET, $11{\beta}$-hydroxy An, $11{\beta}$-hydroxy Et, THS, THA, THE, a-cortolone, a-cortol, $\beta$-cortol, $11{\beta}$-OH Et/$11{\beta}$-OH An and E2/E1 were significantly increased in patients with leiomyoma than in the control group. $17{\beta}$-estradiol was significantly increased in the intramural and the submucosal types than in the subserosal type. There was no significant difference in the concentrations of urinary steroids according to the locations of leiomyomas. There was no significant relationship between the concentration of urinary steroids and the volume of the leiomyomas. $17{\beta}$-estradiol significantly decreased as the weight of uterus increased (r=-0.322, p=0.04). Conclusion: The concentrations of steroid hormone metabolites were generally increased in patients with leiomyoma but were not significantly related to the volume and weight of the leiomyomas. Our study suggests that steroid hormones may be involved in the initiation of leiomyomas but may not be involved in their progression. In addition, the concentrations of steroid hormone metabolites are not related to the leiomyoma type and location.

• Clinical and Experimental Reproductive Medicine
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• 제30권1호
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• pp.23-29
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• 2003

목 적: 본 연구의 목적은 내인성 성호르몬이 자궁근종의 성장에 미치는 영향을 알아보고자 연구를 시행하였다. 연구대상 및 방법: 폐경전 자궁근종 환자 27명과 같은 연령대의 정상여성 25명을 대상으로 하였다. 모든 대상군의 여성에서 24시간 소변을 모아서 소변내 estrogen, androgen의 대사체들을 GC-MS를 이용하여 측정하였으며 두 군에서의 차이를 비교분석 하였다. 결 과: 소변내 $17{\beta}$-estradiol, 5-androstene-$3{\beta}$, $16{\beta}$, $17{\beta}$-triol, 11-keto-ethiocholanolone, $11{\beta}$-hydroxy-androsterone, THS, THA, THE, a-cortolone, a-cortol 및 $\beta$-cortol가 환자군에서 의의있게 증가하였으며 $17{\beta}$estradiol/estrone 및 $11{\beta}$-hydroxy-ethiocholanolone/$11{\beta}$-hydroxy-androsterone도 환자군에서 의의있게 증가하였다. 결 론: 자궁근종의 성장은 요중 estrogen과 androgen의 농도와 밀접한 관련이 있으며 이는 환자의 스테로이드 호르몬 대사 감소에 기인한 것으로 사료된다.

• Asian Pacific Journal of Cancer Prevention
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• 제14권11호
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• pp.6215-6220
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• 2013

Kisspeptins (KPs) encoded by the KiSS-1 gene are C-terminally amidated peptide products, including KP-10, KP-13, KP-14 and KP-54, which are endogenous agonists for the G-protein coupled receptor-54 (GPR54). Functional analyses have demonstrated fundamental roles of KiSS-1 in whole body homeostasis including sexual differentiation of brain, action on sex steroids and metabolic regulation of fertility essential for human puberty and maintenance of adult reproduction. In addition, intensive recent investigations have provided substantial evidence suggesting roles of Kisspeptin signalling via its receptor GPR54 in the suppression of metastasis with a variety of cancers. The present review highlights the latest studies regarding the role of Kisspeptins and the KiSS-1 gene in tumor progression and also suggests targeting the KiSS-1/GPR54 system may represent a novel therapeutic approach for cancers. Further investigations are essential to elucidate the complex pathways regulated by the Kisspeptins and how these pathways might be involved in the suppression of metastasis across a range of cancers.