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小鼠肺泡表面活性物质:通过差速离心制备的亚型的特性

Mouse alveolar surfactant: characterization of subtypes prepared by differential centrifugation.

作者信息

Oulton M, MacDonald J, Janigan D T, Faulkner G T

机构信息

Department of Obstetrics/Gynecology, Dalhousie University, Halifax, Nova Scotia, Canada.

出版信息

Lipids. 1993 Aug;28(8):715-20. doi: 10.1007/BF02535992.

Abstract

To characterize the properties of alveolar surfactant subfractions obtained from mouse lung by differential centrifugation, lavage fluid, following a preliminary centrifugation at 140 x g for 5 min to yield a cellular pellet (Pc), was sequentially centrifuged at 10,000 x g for 30 min, 60,000 x g for 60 min and 100,000 x g for 15 h; and the resultant pellets, respectively referred to as P10, P60 and P100, were harvested for electron microscopy, phospholipid analysis and surface tension measurements. Ultrastructural differences were observed, in that P10 contained large multilamellated structures which were typical of newly secreted surfactant, P100 contained small unilamellar vesicular structures, typical of catabolic end products of alveolar surfactant and P60 appeared to contain a mixture of structures present in P10 and P100 in addition to numerous, large unilamellar vesicles which were not present in either P10 or P100. Slight but significant differences were found in the phospholipid compositions of the three subfractions but not in the fatty acid composition of their phosphatidylcholine (PC) component. There were no significant differences in their disaturated PC/total PC ratios, but significant differences in their phospholipid/protein ratios. P60 had the highest proportion of phospholipid to protein. P10 and P60 demonstrated surface activity but P100 did not. Total alveolar surfactant phospholipid was evenly distributed among the three fractions. This pattern of distribution was significantly different from that observed in rabbit subfractions prepared by the same procedure. These data indicate that mouse alveolar surfactant consists of three distinct subfractions or subtypes which can be separately and quantitatively isolated by differential centrifugation.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

为了表征通过差速离心从鼠肺获得的肺泡表面活性物质亚组分的特性,对灌洗流体在140×g下初步离心5分钟以产生细胞沉淀(Pc)后,依次在10,000×g下离心30分钟、60,000×g下离心60分钟以及100,000×g下离心15小时;所得沉淀分别称为P10、P60和P100,收集用于电子显微镜检查、磷脂分析和表面张力测量。观察到超微结构差异,即P10含有大量多层结构,这是新分泌的表面活性物质的典型特征,P100含有小的单层囊泡结构,这是肺泡表面活性物质分解代谢终产物的典型特征,并且P60似乎除了含有大量P10和P100中均不存在的大单层囊泡外,还含有P10和P100中存在的结构混合物。在这三个亚组分的磷脂组成中发现了轻微但显著的差异,但在其磷脂酰胆碱(PC)组分的脂肪酸组成中未发现差异。它们的二饱和PC/总PC比率没有显著差异,但磷脂/蛋白质比率有显著差异。P60的磷脂与蛋白质比例最高。P10和P60表现出表面活性,但P100没有。肺泡表面活性物质总磷脂在这三个组分中均匀分布。这种分布模式与通过相同程序制备的兔亚组分中观察到的模式显著不同。这些数据表明,鼠肺泡表面活性物质由三个不同的亚组分或亚型组成,可通过差速离心分别和定量分离。(摘要截短于250字)

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