CHARACTERIZATION AND PARTIAL-PURIFICATION OF A FACTOR FROM UREMIC HUMAN-SERUM THAT INDUCES INSULIN RESISTANCE
CHARACTERIZATION AND PARTIAL-PURIFICATION OF A FACTOR FROM UREMIC HUMAN-SERUM THAT INDUCES INSULIN RESISTANCE
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DOI：
10.1172/jci111712
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发表时间：
1985-01-01
影响因子：
15.9
通讯作者：
LOCKWOOD, DH
中科院分区：
文献类型：
--
作者：
MCCALEB, ML;IZZO, MS;LOCKWOOD, DH
正常大鼠脂肪组织与来自非透析、非糖尿病尿毒症患者的血清的孵育在胰岛素存在和不存在的情况下减少葡萄糖的转运和代谢。胰岛素刺激正常大鼠脂肪细胞的葡萄糖代谢被用作生物测定，以鉴定抵抗活性，评估化学修饰对其的影响以及与其产生相关的临床状态。抗性活性是胰蛋白酶不稳定的，并且在6和7之间具有明显的等电点，但不被蛋白A或伴刀豆球蛋白A柱保留。通过与蛋白质合成抑制剂放线菌酮共孵育降低胰岛素抵抗活性。通过加热（100 ℃）获得大于200，000倍的纯化。C）尿毒症血清，将上清液进行Sephadex G-25层析，随后在pH 7.8下吸附到DEAE上，并在pH 6.5下洗脱。部分纯化的抗性活性保留在1，000-MW截留值的透析管内，但不在2000-MW截留值内。血液透析超过1周-18个月的患者显着降低其血清中的抵抗活动的量。在大多数尿毒症患者中存在的抵抗活动，在肾功能正常但肥胖、禁食、老年或患有II型糖尿病的个体的血清中未发现。尿毒症特有的循环小分子量肽通过蛋白质合成依赖性机制诱导胰岛素抵抗。
The incubation of normal rat adipose tissue with sera from nondialyzed, nondiabetic uremic patients reduces the transport and metabolism of glucose, in the absence and presence of insulin. Insulin-stimulated glucose metabolism by normal rat adipocytes was used as a bioassay to identify the resistance activity, assess the effect of chemical modification on it and the clinical states associated with its production. The resistance activity was trypsin-labile and had an apparent isoelectric point between 6 and 7, but was not retained by either protein A or concanavalin A columns. the insulin resistance activity was decreased by coincubation with the protein synthesis inhibitor, cycloheximide. Purification to greater than 200,000-fold was attained by heating (100.degree. C) uremic serum, subjecting the supernatant to Sephadex G-25 chromatography and subsequent adsorption to DEAE at pH 7.8 and elution at pH 6.5. The partially purified resistance activity was retained within dialysis tubing of 1,000-MW cutoff but not within 2000-MW cutoff. Hemodialysis of patients over 1 wk-18 mo reduced significantly the amount of resistance activity in their sera. The resistance activity, present in most uremic patients, was not found in the sera of individuals with normal renal function but who were either obese, fasted, elderly or had type II diabetes mellitus. A circulating small MW peptide, unique to uremia, induced insulin resistance by a protein synthesis-dependent mechanism.