胰高血糖素和胰岛素对成年大鼠肝脏器官培养物中3':5'-环磷酸腺苷浓度的影响。
The effects of glucagon and insulin on adenosine 3':5'-cyclic monophosphate concentrations in an organ culture of mature rat liver.
作者信息
Siddle K, Kane-Maguire B, Campbell A K
出版信息
Biochem J. 1973 Apr;132(4):765-73. doi: 10.1042/bj1320765.
Abstract
- A modified radioimmunoassay for cyclic AMP was developed from the method of Steiner et al. (1969). Cyclic [(3)H]AMP was used as the radioactive tracer. Free and antibody-bound nucleotides were separated by adsorption of protein to Millipore filters. The assay was used to measure amounts of cyclic AMP down to 0.1pmol in 50mul. 2. The effect of glucagon on cyclic AMP content in pieces of mature rat liver maintained for 6 days in organ culture was studied. 3. Cyclic AMP content in the tissue reached a maximum in 5-15min and then decreased. This may have been partly due to an inhibitor of glucagon action formed in the tissue. Small amounts of cyclic AMP were released into the incubation medium. 4. The maximal increase in cyclic AMP content produced by glucagon decreased over 6 days in culture. However, liver pieces cultured for 2 and 6 days were more sensitive to low concentrations of glucagon than were fresh liver pieces. Glucagon concentrations for half-maximal effects were approx. 1mum and 0.05mum for fresh liver and 2-day cultured liver respectively. 5. Insulin (3.5mum) lowered the cyclic AMP content by 30% in the presence of a submaximal glucagon concentration in liver cultured for 2 days. No effect of insulin was demonstrated on fresh liver pieces. 6. Insulin and glucagon were rapidly destroyed by fresh liver pieces.
摘要
- 一种用于环磷酸腺苷（cAMP）的改良放射免疫分析法是在施泰纳等人（1969年）的方法基础上开发的。环[³H]腺苷酸用作放射性示踪剂。通过将蛋白质吸附到微孔滤膜上来分离游离的和与抗体结合的核苷酸。该分析法用于测量50微升中低至0.1皮摩尔的环磷酸腺苷含量。2. 研究了胰高血糖素对在器官培养中维持6天的成年大鼠肝脏切片中环磷酸腺苷含量的影响。3. 组织中的环磷酸腺苷含量在5 - 15分钟内达到最大值，然后下降。这可能部分是由于组织中形成的胰高血糖素作用抑制剂。少量的环磷酸腺苷释放到培养液中。4. 在培养的6天里，胰高血糖素引起的环磷酸腺苷含量的最大增加量下降。然而，培养2天和6天的肝脏切片比新鲜肝脏切片对低浓度的胰高血糖素更敏感。新鲜肝脏和培养2天的肝脏产生半最大效应的胰高血糖素浓度分别约为1微摩尔和0.05微摩尔。5. 在培养2天的肝脏中，在亚最大浓度的胰高血糖素存在下，胰岛素（3.5微摩尔）使环磷酸腺苷含量降低了30%。在新鲜肝脏切片上未显示胰岛素有作用。6. 新鲜肝脏切片能迅速破坏胰岛素和胰高血糖素。
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