Rachmilewitz D, Albers J J, Saunders D R
J Clin Invest. 1976 Feb;57(2):530-3. doi: 10.1172/JCI108307.
We tested whether apoprotein B is present in fasting and postprandial human duodenojejunal mucosa because lipoprotein-like particles are visualized by electron microscopy within the smooth endoplasmic reticulum and the Golgi cisternae of these absorptive cells. Duodenojejunal biopsies from normal volunteers were incubated in citrate buffer and were shaken in 1% EDTA so that absorptive cells could be freed from underlying tissue. Apoprotein B was determined by double-antibody radioimmunoassay in homogenates of absorptive cells. The preparations of absorptive cells were shown to be uncontaminated by plasma lipoproteins; they did not contain any albumin by immunodiffusion able to detect 2 mug/ml. They adsorbed less than 0.1% of 125I-low density lipoprotein which was added to the citrate buffer. Cell preparations from suction biopsies of human rectum contained no detectable apoprotein B. Duodenojejunal absorptive cells from 22 fasting subjects contained 3.2 +/- 0.5 mug of apoprotein B per 100 mg (wet wt) of biopsies or 1.3 mug of apoprotein B per mg of total cell protein. The amount of apoprotein B per milligram of cell protein fell to 0.3 mug in 14 of these individuals whose mucosa was also sampled 45 min after instilling fat intraduodenally. These experiments provide immunochemical evidence that human duodenojejunal absorptive cells contain apoprotein B. This technique should be valuable for studying the physiology of intestinal lipoproteins in absorption and in patients with hyperlipidemia.
由于在这些吸收细胞的滑面内质网和高尔基池中通过电子显微镜可观察到脂蛋白样颗粒,我们检测了载脂蛋白B是否存在于空腹和餐后的人十二指肠空肠黏膜中。取自正常志愿者的十二指肠空肠活检组织在柠檬酸盐缓冲液中孵育,并在1%乙二胺四乙酸(EDTA)中振荡,以便使吸收细胞与下层组织分离。通过双抗体放射免疫分析法测定吸收细胞匀浆中的载脂蛋白B。结果显示,吸收细胞制剂未被血浆脂蛋白污染;通过免疫扩散法检测,其不含任何白蛋白(检测限为2微克/毫升)。它们对添加到柠檬酸盐缓冲液中的125I-低密度脂蛋白的吸附率低于0.1%。取自人直肠吸引活检的细胞制剂中未检测到载脂蛋白B。22名空腹受试者的十二指肠空肠吸收细胞每100毫克(湿重)活检组织含3.2±0.5微克载脂蛋白B,或每毫克总细胞蛋白含1.3微克载脂蛋白B。在其中14名个体中,十二指肠内注入脂肪45分钟后采集其黏膜样本,每毫克细胞蛋白中的载脂蛋白B量降至0.3微克。这些实验提供了免疫化学证据,证明人十二指肠空肠吸收细胞含有载脂蛋白B。这项技术对于研究肠道脂蛋白在吸收过程中的生理学以及高脂血症患者具有重要价值。