Hoff H F, Heideman C L, Gaubatz J W, Gotto A M, Erickson E E, Jackson R L
Circ Res. 1977 Jan;40(1):56-64. doi: 10.1161/01.res.40.1.56.
A quantitiative electroimmunodiffusion (EID) assay was developed for apolipoprotein B (apoB), the major apoprotein of human plasma low density lipoproteins (LDL) and very low density lipoproteins (VLDL). Specificity, sensitivity (30-200 ng) and reproducibility (11%) were established. We used this system to determine the amount of buffer-soluble apoB in supernatant fractions from homogenates of the intima from grossly normal human aortas. Assays of whole tissue minces yielded only one-third of the apoB in supernatant fractions. Intimal homogenates contained 0.34-18.45 mug of apoB/mg of tissue, dry weight (mean 5.42 +/- 3.95 SD). We also found that no apoB was detected in the homogenates of adjacent tunica media. Furthermore, most of the intimal apoB was found in the LDL density (d) fraction (d 1.006-1.063) after differential ultracentrifugation, while the VLDL (d less than 1.006) fraction contained only negligible amounts of apoB. By electron microscopy, it was determined that the LDL density fraction contained particles 200-250 A in diameter which were similar in size to plasma LDL. These results suggest that grossly normal aortas contain significant quantities of intact LDL.
我们开发了一种定量免疫电泳扩散(EID)测定法,用于检测载脂蛋白B(apoB),它是人类血浆低密度脂蛋白(LDL)和极低密度脂蛋白(VLDL)的主要载脂蛋白。该方法的特异性、灵敏度(30 - 200 ng)和重现性(11%)已得到验证。我们使用该系统来测定来自大体正常的人类主动脉内膜匀浆上清液中缓冲液可溶性apoB的含量。对整个组织碎末的测定结果显示,上清液中的apoB含量仅为其三分之一。内膜匀浆中每毫克组织干重含有0.34 - 18.45微克的apoB(平均值为5.42±3.95标准差)。我们还发现,相邻的中膜匀浆中未检测到apoB。此外,在差速超速离心后,大部分内膜apoB存在于LDL密度(d)组分(d 1.006 - 1.063)中,而VLDL(d小于1.006)组分中仅含有少量可忽略不计的apoB。通过电子显微镜观察确定,LDL密度组分中含有直径为200 - 250埃的颗粒,其大小与血浆LDL相似。这些结果表明,大体正常的主动脉含有大量完整的LDL。