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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.