INSULIN-LIKE ACTIVITY OF DILUTE HUMAN SERUM ASSAYED BY AN ISOLATED ADIPOSE CELL METHOD
INSULIN-LIKE ACTIVITY OF DILUTE HUMAN SERUM ASSAYED BY AN ISOLATED ADIPOSE CELL METHOD
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DOI：
10.2337/diab.14.10.643
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发表时间：
1965-01-01
期刊：
影响因子：
7.7
通讯作者：
GLIEMANN, J
中科院分区：
文献类型：
--
作者：
GLIEMANN, J
描述了一种使用分离的脂肪细胞测定胰岛素的方法。在含有葡萄糖（0.1mg/ml）和白蛋白（5%）的Krebs-Ringer碳酸氢盐缓冲液中孵育的脂肪细胞将葡萄糖-1-14 -C转化为CO2。添加胰岛素引起的最大刺激约8至16倍的CO2产生。剂量-反应关系呈典型的S形曲线。最小可检测的胰岛素浓度约为2-5 [mu]U/ml;最大响应在胰岛素浓度约为300 - 600 [mu]U/ml时获得。在最佳范围（约20至160 [mu]U/ml）内，以约[±]10%的精度测定胰岛素浓度（重复4次）。该方法适用于人血清。看来稀释（1%），透析，空腹血清含有大量的“胰岛素样”活性（约20至70 [mu]U/ml）。浓度小于2%的透析血清的剂量反应关系与胰岛素的剂量反应关系相同。随着血清浓度的增加，CO2产生的增量小于随着胰岛素浓度的增加。静脉内葡萄糖负荷后，1%血清的胰岛素样活性没有变化;添加胰岛素抗体不会抑制它。通过淀粉块电泳对血清进行分级，将所有活性定位在[β][中心点] [γ]球蛋白区域的宽峰中。加入血清中的胰岛素在位于白蛋白组分中移动较慢的组分中的单独峰中定量回收。当血清在Sephadex G 75上进行凝胶过滤时，“胰岛素样”活性出现在总排除级分中，而加入血清中的胰岛素在柱上被阻滞。这些性质与先前描述的“非典型”胰岛素的性质相似。其性质仍未确定。
A method for insulin assay using isolated adipose cells is described. Fat cells incubated in Krebs-Ringer bicarbonate buffer containing glucose (0.1 mg/ml) and albumin (5%) converted glucose -1-14 -C to CO2. Addition of insulin caused a maximal stimulation of about 8- to 16-fold of the CO2 production. The dose-response relationship exhibited a typical sigmoid shaped curve. The minimum detectable insulin concentration was about 2-5 [mu]U/ml; the maximum response was obtained with insulin concentrations of about 300 to 600 [mu]U/ml. In the optimal range (about 20 to 160 [mu]U/ml) the insulin concentrations were determined with a precision of about [plus or minus]10% (4 replications). This method was applied to human serum. It appeared that dilute (1%), dialyzed, fasting serum contained large amounts of "insulin-like" activity (about 20 to 70 [mu]U/ml). The dose response relationship of dialyzed serum in a concentration less than about 2% paralleled that of insulin. The increment in CO2 production with greater serum concentration was less than the increment with greater insulin concentrations. The insulin-like activity of 1% serum did not change after an intravenous glucose load; it was not suppressed by addition of insulin antibodies. Fractionation of serum by starch block electrophoresis located all of the activity in a broad peak in the [beta][center dot] [gamma] globulin region. Insulin added to serum was quantitatively recovered in a separate peak located in the slower moving component of the albumin fraction. When serum was subjected to gel filtration on Sephadex G 75 the "insulin-like" activity emerged with the total excluded fraction whereas insulin added to serum was retarded on the column. These properties were similar to those of the previously described "atypical" insulin. Its nature remained undefined.