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스테로이드 합성을 교란하는 내분비계장애물질 검색을 위한 라이디히 세포 분리 및 배양조건 확립
강일현(Il Hyun Kang),강태석(Tae Seok Kang),강호일(Ho Il Kang),문현주(Hyun Ju Moon),김태성(Tae Sung Kim),기호연(Ho Hyun Ki),류혜원(Hye Won Ryu),신재호,동미숙(Mi Sook Dong),한순영(Soon Young Han),김승희(Seung Hee Kim),홍진환(Jin Hwan Hong 한국환경성돌연변이발암원학회 2006 한국환경성돌연변이·발암원학회지 Vol.26 No.2
Normally, environmental toxicants are classified as endocrine disruptors if they interfere with regulation of cellular function by endogeneous steroids through inhibition of receptor binding and/or transcriptional activation. So, many studies have been performed about agonist/antagonist of hormone receptor to study mechanisms of endocrine disruptors. If toxicants affect steroid biosynthesis and/or degradation and alter hormone homeostasis, these also are classified as endocrine disruptors. But there are not many studies of the mechanisms of endocrine disruptors on the basis of alteration of steroid biosynthesis and/or degradation. Isolation and culture of Leydig cells from testis is one of methods for the steroidogenesis screening assays to evaluate a substance for altering steroidogenesis. Leydig cells were harvested using the method described by Klinefelter with modifications. Leydig cells were purified by perfusion of testis and incubation (34℃, 80cycles/minute, 20 minutes) with collagenase (0.25 ㎎/㎏), centrifugal elutriation, percoll gradient centrifugation and BSA multidensity gradient centrifugation. To confirm if this method is one of appropriate tools to evaluate a substance for altering steroidogenesis, ketoconazole, positive control was administered to purified Leydig cells. Ketoconazole (10??M and above) significantly reduced testosterone production in purified Leydig cells. From above results, we suggest that this method for steroidogenesis screening assay appears to be a appropriate tool to detect suspected compounds for altering steroidogenesis.
LC-MS/MS를 이용한 인체시료 중 프탈레이트 대사체 동시분석법 확립
홍순근,남혜선,정기경,강일현,김태성,조상은,정수희,이장우,김준철,고영림,강태석,Hong, Soon-Keun,Nam, Hye-Seon,Jung, Ki-Kyung,Kang, Il-Hyun,Kim, Tae-Sung,Cho, Sang-Eun,Jung, Su-Hee,Lee, Jang-Woo,Kim, Jun-Cheol,Kho, Young-Lim,Kang, Tae-Se 한국환경보건학회 2010 한국환경보건학회지 Vol.36 No.6
Phthalates, such as di (2-ethylhexyl) phthalate (DEHP), dibutyl phthalate (DBP) have been proved to be teratogenics and endocrine disruptors, metabolized rapidly and excreted in the urine. In this study, a simultaneous analytical method for 10 phthalate metabolites, MnBP, MiBP, MBzP, MCHP, MEHP, MEHHP, MEOHP, MnOP, MiNP and MiDP, in human urines, based on switching system with on-line pretreatment column using HPLC-MS/MS has been developed. This method was validated according to the guideline of bioanalytical method validation of National Institute of Toxicological Research. Limits of detection range between 0.2 and 0.9 ng/ml for 10 phthalate metabolites. The calibration curves showed linearity in the range 0.997~0.999, and the results of the intra- and inter-day validations were in the range from 0.4 to 14.7% RSD and from 0.3 to 9.4% RSD, respectively. Recoveries of phthalate metabolites varied from 87.0 to 116.1%. This analytical method showed high accuracy and stable precision for all metabolites, and seems to be suitable for biomonitoring of phthalates in human urine.
항안드로겐성 물질이 성 성숙 이전 단계의 정소에서 미치는 영향 연구
홍진,한순영,문현주,강태석,강일현,김태성,김승희,권기성,Hong Jin,Han Soon-Young,Moon Hyun-Ju,Kang Tae-Seok,Kang Il-Hyun,Kim Tae-Sung,Kim Seung-Hee,Kwon Ki-Sung 환경독성보건학회 2006 환경독성보건학회지 Vol.21 No.3
The experiments investigated whether early exposure to testosterone propionate (TP) during prepuberty alters testis development in Sprague-Dawley male rats. We performed Hershberger assay using the stimulated weanling male rats by OECD protocols, cDNA microarray, and Western blot. TP was subcutaneously injected to uncastrated Sprague-Dawley male rat of 22 days old for 10 consecutive days at doses of 0.4, 0.8, 1.0, 1.2, 1.6 mg/kg per day. At necropsy, the following tissues were removed and weighed: combined testes, epididymides (Epi), Cowper's glands (COW), levator am, and bulbocavernosus muscles (LABC), seminal vesicles, together with coagulating gland (SV) and ventral prostate (VP). We found that TP increased the weights of Epi, VP, SV, COW, and LABC, while testis was decreased in a dose-dependent manner. In cDNA microarray analysis of testis, there were significant reductions in the expression of cytochrome P450 11A (CYP11A), the rate-limiting enzyme of steroidogenesis. Taken together these results, TP exposure before puberty in male rats may produce the delay in testis development by inhibiting the CYP11A gene expression.