Parker M T, Turrentine M A, Johnson G S
Department of Veterinary Pathology, College of Veterinary Medicine, University of Missouri, Columbia 65211.
Am J Vet Res. 1991 Jan;52(1):119-25.
Canine and human platelets (washed 4 times in a solution containing EDTA, prostaglandin E1, and theophylline to prevent release of alpha-granule constituents) were lysed by being frozen and thawed in the presence of detergent. Radioelectroimmunoassay for von Willebrand factor (vWf) in 5 human platelet lysates produced precipitin rockets, shaped like those produced from vWf in plasma from healthy human beings, and indicated that the mean von Willebrand factor antigen (vWf:Ag) content in platelets from healthy human being was 526 +/- 87 human U/10(12) platelets. Radioelectroimmunoassay for vWf in platelet lysates from 17 healthy dogs with normal plasma. vWf:Ag concentration produced precipitin rockets that looked different from those produced from canine plasma and indicated vWf:Ag content of 59 +/- 35 canine U/10(12) platelets. Inclusion of protease inhibitors in the lysing solution did not normalize the appearance of the precipitin rockets or substantially alter the measured platelet content of vWf:Ag. The array of vWf multimers revealed by sodium dodecyl sulfate-agarose gel electrophoresis of canine platelet lysates had a distinct appearance that differed from that of vWf in canine or human plasma and platelets; the intensity of the canine platelet vWf multimer bands was skewed, with relatively greater density in the lower molecular weight region and faint or undetectable multimer bands in the higher molecular weight region. Electrophoretograms with visible multimers in the high molecular weight region had vWf components that had higher molecular weight than did any vWf components in canine plasma. Radioelectroimmunoassay for fibronectin in these same canine platelet lysates indicated that the fibronectin content in platelets was 2.89 +/- 1.10 mg/10(12) platelets.(ABSTRACT TRUNCATED AT 250 WORDS)
犬类和人类血小板（在含有乙二胺四乙酸、前列腺素E1和茶碱的溶液中洗涤4次，以防止α-颗粒成分释放）在去污剂存在的情况下通过冻融进行裂解。对5份人类血小板裂解物进行的血管性血友病因子（vWf）放射免疫电泳分析产生了沉淀火箭，其形状与健康人血浆中的vWf产生的沉淀火箭相似，表明健康人血小板中血管性血友病因子抗原（vWf:Ag）的平均含量为526±87人类单位/10¹²个血小板。对17只血浆正常的健康犬的血小板裂解物进行vWf放射免疫电泳分析。vWf:Ag浓度产生的沉淀火箭与犬血浆产生的沉淀火箭不同，表明vWf:Ag含量为59±35犬单位/10¹²个血小板。在裂解溶液中加入蛋白酶抑制剂并没有使沉淀火箭的外观正常化，也没有显著改变所测的血小板vWf:Ag含量。通过十二烷基硫酸钠-琼脂糖凝胶电泳显示的犬血小板裂解物中的vWf多聚体阵列具有独特的外观，与犬或人类血浆及血小板中的vWf不同；犬血小板vWf多聚体条带的强度呈偏态分布，在较低分子量区域密度相对较高，在较高分子量区域条带较淡或无法检测到。在高分子量区域有可见多聚体的电泳图谱中，vWf成分的分子量高于犬血浆中的任何vWf成分。对这些相同犬血小板裂解物中的纤连蛋白进行放射免疫电泳分析表明，血小板中的纤连蛋白含量为2.89±1.10毫克/10¹²个血小板。（摘要截于250字）
