Hulstaert C E, Gijzel W P, Hardonk M J, Kroon A M, Molenaar I
Lab Invest. 1975 Aug;33(2):176-86.
The objective of this investigation was to find out whether vitamin E deficiency, apart from influencing the lipid component of cellular membranes, also influences the protein component. For that purpose a number of membrane-bound enzymes in the liver of the Pekin duckling were histochemically, cytochemically, and biochemically examined. Furthermore, cells, cellular membranes, and protein particles in membranes were morphometrically investigated. Histochemically five membrane-bound enzymes appeared to be stimulated in vitamin E deficiency: 5'-nucleotidase, glucose-6-phosphatase, isocitrate dehydrogenase (NADP), tetrazolium reductase (NADH), and tetrazolium reductase (NADPH). 5'-Nucleotidase and glucose-6-phosphatase were also investigated cytochemically and biochemically. The cytochemical localization of these enzymes was identical in control and vitamin E-deficient ducklings. Biochemically, a stimulation of these two enzymes also could be demonstrated. The increase per milligram of DNA appeared to be largest whereas the increase per milligram of protein, per milligram of phospholipid, and per milligram of RNA was only half of the increase per milligram of DNA. This can be explained by the 30 per cent increase of the cell volume in vitamin E deficiency leading to an increase of protein, phospholipid, and RNA per cell. The thickness of membranes and the diameter of protein particles in membranes were measured in liver parenchymal cells. In vitamin E deficiency the thickness of the outer mitochondrial membrane and the diameter of protein particles in this membrane were smaller whereas the thickness of the endoplasmic reticular membrane was larger. The increase of the activities of mitochondrial and microsomal enzymes and the decrease of the thickness of the outer mitochondrial membrane and of its protein particles are interpreted to be the result of the influence of free radicals on membranes with electron transport functions. The increase of 5'-nucleotidase activity in the plasma membrane is likely to have a different cause; it may be related to the transport of nucleotides across this membrane.
本研究的目的是探讨维生素E缺乏除了影响细胞膜的脂质成分外,是否还会影响蛋白质成分。为此,对北京鸭雏鸭肝脏中的多种膜结合酶进行了组织化学、细胞化学和生物化学检测。此外,还对细胞、细胞膜和膜中的蛋白质颗粒进行了形态计量学研究。组织化学研究表明,维生素E缺乏时,五种膜结合酶似乎受到刺激:5'-核苷酸酶、葡萄糖-6-磷酸酶、异柠檬酸脱氢酶(NADP)、四氮唑还原酶(NADH)和四氮唑还原酶(NADPH)。还对5'-核苷酸酶和葡萄糖-6-磷酸酶进行了细胞化学和生物化学研究。在对照鸭和维生素E缺乏鸭雏鸭中,这些酶的细胞化学定位相同。生物化学研究也证实了这两种酶受到刺激。每毫克DNA的增加量似乎最大,而每毫克蛋白质、每毫克磷脂和每毫克RNA的增加量仅为每毫克DNA增加量的一半。这可以用维生素E缺乏时细胞体积增加30%来解释,这导致每个细胞中的蛋白质、磷脂和RNA增加。测量了肝实质细胞中膜的厚度和膜中蛋白质颗粒的直径。维生素E缺乏时,线粒体外膜的厚度和该膜中蛋白质颗粒的直径较小,而内质网膜的厚度较大。线粒体和微粒体酶活性的增加以及线粒体外膜及其蛋白质颗粒厚度的减小被解释为自由基对具有电子传递功能的膜产生影响的结果。质膜中5'-核苷酸酶活性的增加可能有不同的原因;它可能与核苷酸跨该膜的转运有关。