de Swart C A, Nijmeyer B, Andersson L O, Holmer E, Verschoor L, Bouma B N, Sixma J J
Blood. 1984 Apr;63(4):836-42.
High and low affinity heparin (HA and LA heparin) were prepared from commercial heparin by affinity chromatography to insolubilized antithrombin III. HA heparin was radiolabeled with 35S and subdivided by gel chromatography into high molecular weight (HMW, average 17,000-26,000 daltons), intermediate molecular weight (MMW, average 12,000-13,000 daltons), low molecular weight (LMW, average 5,000-7,000 daltons), and very low molecular weight (VLMW, average 4,600 daltons) fractions. The kinetics of lipolytic and anticoagulant activity and protein-bound radioactivity were studied after intravenous injection of these fractions. LA heparin failed to induce anticoagulant activity but released the hepatic triglyceride lipase (H-TGL) and lipoprotein lipase (LPL) activities normally. VLMW and LMW heparin failed to release both lipolytic enzymes and did not induce anticoagulant activity measurable by the activated partial thromboplastin time (APTT). A powerful anticoagulant effect was found in the anti-Xa assay, which disappeared according to a continuously concave curve in semilogarithmic plots, with elimination rates similar to those of the protein-bound radiolabel. The other heparin preparations induced all activities measured. Heparin anticoagulant activity estimated by the two assays disappeared following a convex curve, preceded by a rapid initial elimination phase in semilogarithmic plots. The disappearance rates of plasma protein-bound heparin radioactivity and heparin anticoagulant activity estimated by factor Xa inactivation were similar. Peak values of the two lipolytic activities were attained rapidly. H- TGL activity, as well as LPL activity, disappeared following convex curves in semilogarithmic plots, with elimination rates similar to those of plasma protein-bound heparin radioactivity. On the basis of these kinetics, we suggest that, after intravenous administration of heparin, the two lipolytic enzymes present in plasma are complexed with heparin, analogous to the heparin-antithrombin III complex. Finally, the kinetic data indicate that elimination of these activities is determined by the heparin part of the complexes, probably by removal of free heparin.
通过亲和色谱法将市售肝素与固定化抗凝血酶III结合，制备出高亲和力肝素（HA肝素）和低亲和力肝素（LA肝素）。用35S对HA肝素进行放射性标记，然后通过凝胶色谱法将其细分为高分子量（HMW，平均17,000 - 26,000道尔顿）、中分子量（MMW，平均12,000 - 13,000道尔顿）、低分子量（LMW，平均5,000 - 7,000道尔顿）和极低分子量（VLMW，平均4,600道尔顿）组分。静脉注射这些组分后，研究了脂解活性、抗凝活性和蛋白质结合放射性的动力学。LA肝素未能诱导抗凝活性，但能正常释放肝甘油三酯脂肪酶（H-TGL）和脂蛋白脂肪酶（LPL）的活性。VLMW和LMW肝素未能释放这两种脂解酶，也未诱导通过活化部分凝血活酶时间（APTT）可测量的抗凝活性。在抗Xa测定中发现了强大的抗凝作用，在半对数图中，该作用根据连续的凹曲线消失，消除速率与蛋白质结合放射性标记的消除速率相似。其他肝素制剂诱导了所有测量的活性。通过两种测定法估计的肝素抗凝活性在凸曲线后消失，在半对数图中，之前有一个快速的初始消除阶段。血浆蛋白结合肝素放射性和通过因子Xa失活估计的肝素抗凝活性的消失速率相似。两种脂解活性的峰值迅速达到。H-TGL活性以及LPL活性在半对数图中遵循凸曲线消失，消除速率与血浆蛋白结合肝素放射性的消除速率相似。基于这些动力学，我们认为，静脉注射肝素后，血浆中存在的两种脂解酶与肝素形成复合物，类似于肝素 - 抗凝血酶III复合物。最后，动力学数据表明，这些活性的消除由复合物中的肝素部分决定，可能是通过去除游离肝素。
