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放射性标记和胶体金标记的乳糜乳糜微粒及乳糜微粒残粒在体外被大鼠血小板摄取的情况。

Uptake of radiolabeled and colloidal gold-labeled chyle chylomicrons and chylomicron remnants by rat platelets in vitro.

作者信息

Xu N, Zhou L, Odselius R, Nilsson A

机构信息

Department of Cell Biology 1, University Hospital of Lund, Sweden.

出版信息

Arterioscler Thromb Vasc Biol. 1995 Jul;15(7):972-81. doi: 10.1161/01.atv.15.7.972.

Abstract

This study examined the uptake of chyle chylomicrons (CMs) and chylomicron remnants (CMRs) by rat platelets in vitro. CMs and CMRs were doubly labeled with [3H]arachidonate ([3H]-20:4) and [14C]cholesterol and were incubated with platelets for up to 4 hours. A significant uptake (binding and/or internalization) of CMs by the platelets occurred, as indicated by the parallel increase of [3H]20:4 and [14C]cholesterol in platelets with incubation time. Addition of unlabeled CMs, VLDLs, LDLs, and HDLs decreased the uptake of labeled CMs. The competition experiments suggested that there is both a saturable binding and a nonspecific uptake of CMs. During incubation with CMs, the proportion of [3H]20:4 in phospholipids decreased and that in 1,2-x-diacylglycerol increased. The data indicated that a phospholipase C-mediated degradation of phosphatidylcholine and phosphatidylethanolamine occurred, whereas [3H]20:4 in triglyceride and 14C in cholesterol ester did not change. Electron microscopic studies after incubation with colloidal gold-labeled CMs (CM-Au's) demonstrated an accumulation of CM-Au particles in the open canalicular system of the platelets. Some CM-Au particles were localized in cytoplasmic vacuoles that were not stained by ruthenium red. Some CM-Au's or free gold particles were in vacuoles that showed acid phosphatase activity, indicating that some true endocytosis of CM occurred. The uptake of [3H]-20:4- and [14C]cholesterol-labeled CMRs was low compared with the uptake of CMs. After incubation with colloidal gold-labeled CMRs (CMR-Au's), only a few platelets contained CMR-Au in their open canalicular systems, and no CMR-Au particles were seen in the cytoplasm or in acid phosphatase-positive vacuoles. Rat platelets can thus interact with CMs by a process that leads to a sequestration in the open canalicular system and endocytosis and a net degradation of CM phospholipids. The conversion of CMs to CMRs counteracts this interaction.

摘要

本研究在体外检测了大鼠血小板对乳糜微粒(CMs)和乳糜微粒残粒(CMRs)的摄取情况。CMs和CMRs用[3H]花生四烯酸([3H]-20:4)和[14C]胆固醇进行双重标记,并与血小板孵育长达4小时。血小板对CMs有显著摄取(结合和/或内化),这表现为随着孵育时间的延长,血小板中[3H]20:4和[14C]胆固醇平行增加。添加未标记的CMs、极低密度脂蛋白(VLDLs)、低密度脂蛋白(LDLs)和高密度脂蛋白(HDLs)会降低标记CMs的摄取。竞争实验表明,CMs存在可饱和结合和非特异性摄取。在与CMs孵育期间,磷脂中[3H]20:4的比例降低,而1,2-二酰基甘油中的比例增加。数据表明发生了磷脂酶C介导的磷脂酰胆碱和磷脂酰乙醇胺降解,而甘油三酯中的[3H]20:4和胆固醇酯中的14C没有变化。用胶体金标记的CMs(CM-Au's)孵育后的电子显微镜研究表明,CM-Au颗粒在血小板的开放小管系统中积累。一些CM-Au颗粒位于未被钌红染色的细胞质空泡中。一些CM-Au's或游离金颗粒存在于显示酸性磷酸酶活性的空泡中,这表明发生了一些CM的真正内吞作用。与CMs相比,[3H]-20:4和[14C]胆固醇标记的CMRs的摄取较低。用胶体金标记的CMRs(CMR-Au's)孵育后,只有少数血小板在其开放小管系统中含有CMR-Au,在细胞质或酸性磷酸酶阳性空泡中未见CMR-Au颗粒。因此,大鼠血小板可通过一种导致在开放小管系统中隔离、内吞以及CM磷脂净降解的过程与CMs相互作用。CMs向CMRs的转化会抵消这种相互作用。

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