• 作者： 林仁壽; 謝美華; 林文翰
• 作者服務機構： 臺灣大學畜牧系; 榮民總醫院臺中分院新陳代謝科
• 中文摘要： 本試驗之目的在改良現有的分離腎上腺細胞法以供微量鼠血漿之促腎上皮素測定。所使用的改良方法包括修整(priming), Methyl-isobutylxanthsine或dexamethasone處理。由結果顯示僅有dexamethasone能得到預期的效果。 大鼠先以dexamethasone(100 g/kg BM.)腹腔注射，一小時經迅速斷頭犧牲，取出腎上腺，去除膜鞘，切成8-10片，再以 Collagenase 及 deoxyribonuclease 消化三十分鐘，即可得到分離之細胞，將此細胞以 Krebs-Ringer溶液(KR-BGA)洗淨，並分裝於小試管中，每試管含3~4x 10 細胞，再加入促腎上皮素(ACTH)標準液或血漿抽出液，於37°C下培育2小時，並通入 95％ O 及 5％ CO 。然後以螢光比色怯測定此混合液中腎上皮質酮 (Corticosterone)之含量，以混合液中腎上皮質酮的總產量來預估血漿中的促腎上皮素的含量。本法極為精確 ( =0.06)，靈敏度(10 fg)及便捷。通常以10～15頭大鼠為材料，一位熟練的人員，一日可以分析15～20血樣，而血漿抽出物僅需 0.01～0.05毫升。
• 英文摘要： The present study was designed to improve the dispersed adrenal cell technique for determiningadrenocorticotrophic hormone (ACTH) concentrations in small amounts of rat plasma. Priming withACTH, incubation with methyl-isobutlyxanthine, or dexamethasone pre-treatment were employed as modi-fications. Of these, only dexamethasone pre-treatment increased the sensitivity of the assay. The adrenal fragments obtained from 10-12 adult male rats pre-treated with dexamethasone (100 g/kg B.W.) one hour before sacrifice, were digested with collagenase and deoxyribonuclease solution for30 minutes. The dispersed cells were collected by centrifugation and resuspended in Krebs-Ringer bi-carbonate buffer containing 0.2% glucose and 0.5% bovine serum albumin. Aliquots of cell suspension(3-4x10 /tube) were incubated with various doses of ACTH or the eluate of plasma samples at 37°Cfor 2 hours in an atmosphere of 95% O /5% CO in a Dubnoff shaker. The quantity of corticosteroneproduced was measured fluorimetrically. The assay is precise ( = 0.06), extremely sensitive (10 fg/tube),and convenient. One skilled technician can handle 15 to 20 plasma samples per day using 10 rats as thesource of assay cells. ACTH can be measured in as little as 10-50 l of eluate.
• 中文關鍵字： adrenal cell; corticostropin; bioassay; rat
• 英文關鍵字： --