Kitteringham N R, Kenna J G, McLean C, Clarke J B, Park B K
Department of Pharmacology and Therapeutics, University of Liverpool, UK.
Drug Metab Dispos. 1992 Sep-Oct;20(5):625-31.
The extent of protein dinitrophenylation was determined in plasma and other tissues of anesthetized rats after administration of the model immunogen [3H]dinitrofluorobenzene (DNFB) (25 mg/kg; 5-25 microCi). DNFB was given by the intravenous, intraportal, intramuscular, or oral route. Irreversible binding was determined radiometrically after exhaustive solvent extraction of plasma or organ proteins. The extent of binding was high in plasma after parenteral administration (approximately 1% dose/ml plasma), but less (approximately 0.1% dose/ml) if DNFB was given orally. Low levels of radioactivity were bound irreversibly in liver (0.01-0.13% dose/g) and kidney (0.03-0.10% dose/g) and only residual amounts in other organs. Western blotting was used to identify target proteins in plasma, liver, and kidney using a specific antidinitrophenyl antiserum. No dinitrophenylation could be detected in liver or kidney samples, but strong recognition of two protein bands was observed in plasma. Bands with the same apparent molecular masses (67 and 44 kDa) were seen when DNFB was incubated with rat plasma in vitro. Preliminary evidence for these proteins being albumin and alpha 1-acid glycoprotein, respectively, is presented. The latter may be important for interindividual variability in immune responsiveness, because it is an acute phase protein whose levels fluctuate widely during disease states.
在给麻醉大鼠注射模型免疫原[3H]二硝基氟苯（DNFB）（25mg/kg；5 - 25微居里）后，测定血浆和其他组织中蛋白质二硝基苯化的程度。DNFB通过静脉、门静脉、肌肉或口服途径给药。在对血浆或器官蛋白质进行彻底的溶剂萃取后，通过放射性测定法确定不可逆结合情况。肠胃外给药后血浆中的结合程度较高（约1%剂量/毫升血浆），但口服DNFB时结合程度较低（约0.1%剂量/毫升）。肝脏（0.01 - 0.13%剂量/克）和肾脏（0.03 - 0.10%剂量/克）中不可逆结合的放射性水平较低，其他器官中仅有残留量。使用特异性抗二硝基苯抗血清通过蛋白质印迹法鉴定血浆、肝脏和肾脏中的靶蛋白。在肝脏或肾脏样本中未检测到二硝基苯化，但在血浆中观察到对两条蛋白带的强烈识别。当DNFB在体外与大鼠血浆孵育时，可见具有相同表观分子量（67和44 kDa）的条带。分别提出了这些蛋白质可能是白蛋白和α1 - 酸性糖蛋白的初步证据。后者可能对个体间免疫反应性的差异很重要，因为它是一种急性期蛋白，其水平在疾病状态期间波动很大。
