PATHOBIOLOGICAL DETERMINANTS OF ATHEROSCLEROSIS IN YOUTH

此申请是以美国参与单位的身份提交的 研究人员发起的病理生物学多中心研究 青年动脉粥样硬化的决定因素。的概念基础 应用取决于几个考虑和观察。名列前茅 这些是：1)主要血管壁的功能完整性是， 在大多数情况下，通过胶原纤维的存在来维持； 2)胶原蛋白的含量及各组分的相对比例 胶原蛋白的类型通常在导致 人类动脉粥样硬化病变的表现。此外，它是 认为导致动脉粥样硬化形成的过程 病变可以在较长的时间间隔内手术。在这方面， 参与预期的财团安排提供了一个独特的 评估血管胶原蛋白状态的潜在修饰的机会 在确定的临床前人群中，并将发现与年龄相关， 总体研究包括受试者的性别、种族和栖息地。 因此，预计参与该项目的收集中心 这项研究每年将提供大约50个血管组织样本。 样本将来自25名受试者，涉及一名解剖学定义的 胸主动脉节段(20毫米)以及一个解剖定义的 每个受试者左回旋支冠状动脉的节段(40毫米)。这个 预期分析的复杂性排除了分析不仅仅是 按年注明样品。关于分析，每一项 样本将被去除外膜，通过物理手段进行精细划分，并 对小份进行氨基酸分析，以评估总含量 胶原蛋白含量。因为之前的调查表明，有限的 胃酶对蛋白质的降解在数量上只有轻微的效果 溶解血管组织中的胶原蛋白，胶原蛋白将 利用特定的化学裂解将其溶解为多肽片段 CNBR。然后将得到的多肽片段进行层析 CM-纤维素和部分已知含有标记物的柱状流出物 从I型、III型、IV型和V型胶原链衍生的多肽将是 选择凝胶渗透高效液相色谱法进行进一步分析，以实现 来自不同链的多肽的拆分和定量。 由于I、III、IV和V型胶原是主要的胶原蛋白 在血管组织中，这些分析将提供基本数据 关于任何潜在的数量和/或质量变化 血管胶原蛋白状态作为上述参数的函数。

This application is submitted as a participating unit in the USA investigator-initiated multicenter study of the pathobiological determinants of atherosclerosis in youth. The conceptual basis for the application rests on several considerations and observations. Chief among these are: 1) the functional integrity of the walls of major vessels is, for the most part, maintained through the presence of collagen fibers; and 2) the quantity of collagen as well as the relative proportions of various collagen types is generally altered during the processes leading to the appearance of atherosclerotic lesions in humans. In addition, it is believed that the processes leading to the formation of atherosclerotic lesions are operative over lengthy intervals. In this regard, participation in the contemplated consortium arrangement provides a unique opportunity to evaluate potential modifications of vascular collagen status in a defined preclinical population and correlate the findings with age, sex, race and habitat of the subjects included in the overall study. Accordingly, it is anticipated that the collecting centers involved in the study will provide approximately 50 samples of vascular tissue per year. The samples will be from 25 subjects and involve one anatomically-defined segment (20 mm) of thoracic aorta as well as one anatomically defined segment (40 mm) of the left circumflex coronary artery per subject. The complexity of the contemplated analyses precludes analyses of more than the indicated samples on a yearly basis. With respect to the analyses, each sample will be freed of adventitia, finely divided by physical means, and a small aliquot subjected to amino acid analysis in order to evaluate total collagen content. Since previous investigations have shown that limited proteolysis with pepsin is only marginally effective in quantitatively solubilizing collagens in vascular tissue, the collagens will be solubilized as peptide fragments utilizing specific chemical cleavage with CNBr. The resulting peptide fragments will then be chromatographed on CM-cellulose and portions of the column effluent known to contain marker peptides derived from the chains of type I, III, IV and V collagens will be chosen for further analyses by gel permeation HPLC in order to achieve resolution and quantitation of the peptides derived from various chains. Since type I, III, IV and V collagens constitute the major collagens present in vascular tissues, these analyses will provide basic data concerning any potential quantitative and/or qualitative alterations in vascular collagen status as a function of the parameters noted above.