Nejjar I, Pieraggi M T, Thiers J C, Bouissou H
Service d'Anatomie Pathologique, C.H.U., Toulouse, France.
Atherosclerosis. 1990 Jan;80(3):199-208. doi: 10.1016/0021-9150(90)90027-g.
Thirty human aortas with varying degrees of atheroma graded macroscopically according to the WHO classification were taken at autopsy from subjects of different ages (24-86 years). Study by light microscopy showed aortas with an intact wall (4 subjects, 25-46 years) with a thin intima and regular elastic layers, and aortas with varying degrees of modification of the wall, where the intima was of varying thickness and the elastic fibers showed varying degrees of damage (moderate lesions: 5 subjects, 35-52 yrs; severe lesions: 21 subjects, 26-86 yrs). From each aorta, a 4-cm segment from the tunica media, free of atheromatous lesions, was defatted and subjected to successive treatment with EDTA-Tris, 6 M guanidine-HCl-Tris, 6 M guanidine-HCl-Tris-DTE and collagenase. The residues (EP residues) were subjected to amino acid (AA) analysis and transmission electron microscopy (TEM) study. In the young subject, the AA composition was similar to that of elastin and the TEM images were characteristic of this substance. In the aging subject, an increase in polar AA and a parallel decrease in apolar AA and crosslinks was noted. By TEM, the elastin was seen to be associated with abundant fibrillar material. Trypsin treatment of EP residues gave E residues, whose composition and TEM appearance were similar in all samples, corresponding to the standard composition of elastin and its classic appearance by electron microscopy. We suggest that the fibrillar material removed by trypsin is the morphological reflection of the chemical variations observed in the EP residues. These correspond to contamination of the elastin by a polar protein fraction. This contamination is closely correlated with age but not with the degree of atheroma. Thus the age-related chemical changes in elastin appear to be independent of the onset and evolution of atheromatous lesions. The 10-15 nm diameter of the contaminating fibrillar material suggests that may be the microfibrillar fraction of elastic tissue.
从不同年龄(24 - 86岁)的受试者尸检中获取30个根据世界卫生组织分类进行宏观分级的不同程度动脉粥样硬化的人体主动脉。光镜研究显示,主动脉壁完整的(4名受试者,25 - 46岁)内膜薄且弹性层规则,以及主动脉壁有不同程度改变的情况,其中内膜厚度各异且弹性纤维有不同程度损伤(中度病变:5名受试者,35 - 52岁;重度病变:21名受试者,26 - 86岁)。从每个主动脉中取一段来自中膜的4厘米无动脉粥样病变的节段,进行脱脂处理,并依次用乙二胺四乙酸 - 三羟甲基氨基甲烷(EDTA - Tris)、6M盐酸胍 - 三羟甲基氨基甲烷、6M盐酸胍 - 三羟甲基氨基甲烷 - 二硫苏糖醇(DTE)和胶原酶处理。将残余物(EP残余物)进行氨基酸(AA)分析和透射电子显微镜(TEM)研究。在年轻受试者中,氨基酸组成与弹性蛋白相似,TEM图像具有该物质的特征。在老年受试者中,发现极性氨基酸增加,非极性氨基酸和交联键平行减少。通过TEM观察到弹性蛋白与大量纤维状物质相关。用胰蛋白酶处理EP残余物得到E残余物,其组成和TEM外观在所有样品中相似,与弹性蛋白的标准组成及其电子显微镜下的经典外观相对应。我们认为,胰蛋白酶去除的纤维状物质是在EP残余物中观察到的化学变化的形态学反映。这些变化对应于弹性蛋白被极性蛋白质部分污染。这种污染与年龄密切相关,但与动脉粥样硬化程度无关。因此,弹性蛋白中与年龄相关的化学变化似乎与动脉粥样硬化病变的发生和发展无关。污染性纤维状物质的直径为10 - 15纳米,表明其可能是弹性组织的微纤维部分。