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分离灌注肺的II型细胞对表面活性物质磷脂的摄取和再利用

Uptake and reutilization of surfactant phospholipids by type II cells of isolated perfused lung.

作者信息

Moxley M A, Jacoby J, Longmore W J

机构信息

E.A. Doisy Department of Biochemistry and Molecular Biology, St. Louis University School of Medicine, Missouri 63104.

出版信息

Am J Physiol. 1991 Apr;260(4 Pt 1):L268-73. doi: 10.1152/ajplung.1991.260.4.L268.

Abstract

To investigate the role of type II alveolar cells in surfactant uptake and reutilization while cells are in situ, the combination of isolated perfused rat lung and type II cell isolation has been used. After an equilibration period, isolated perfused rat lungs received an intratracheal injection of labeled rat surfactant or a labeled bovine-derived preparation, and the perfusion continued. At time intervals less than or equal to 3 h, lungs were lavaged and type II cells isolated. Freshly isolated cells from perfused lungs were highly viable as judged by trypan blue exclusion and by linear incorporation of labeled leucine into trichloroacetic acid precipitable protein during perfusion. After cell isolation, total phospholipid or phosphatidylcholine was extracted from cells. Incorporation of surfactant phosphatidylcholine label into cellular phosphatidylcholine was shown to be linear with time whether lungs received natural rat surfactant or bovine-derived surfactant preparation. Uptake of natural surfactant from alveoli into type II cells was approximately three times greater than uptake of bovine-derived material. Results of administration of 0.5 mumol surfactant phospholipid or 2.0 mumol were similar except that administration of 2.0 mumol of natural rat surfactant caused uptake to become saturated at 1 h of perfusion. An increase in cell-associated phosphatidylcholine occurred whether results were expressed on basis of cellular DNA or specific activity of cellular phosphatidylcholine. Majority of administered surfactant label remained lavage fluid associated at 3 h, whereas remainder was associated with isolated cells or debris from cell preparation. Little label was detected in perfusion medium.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

为了研究Ⅱ型肺泡细胞在原位时对表面活性剂摄取和再利用的作用,采用了分离灌注大鼠肺脏与Ⅱ型细胞分离相结合的方法。在平衡期后,对分离灌注的大鼠肺脏进行气管内注射标记的大鼠表面活性剂或标记的牛源性制剂,然后继续灌注。在小于或等于3小时的时间间隔内,对肺脏进行灌洗并分离Ⅱ型细胞。从灌注肺脏中新鲜分离的细胞具有很高的活力,这通过台盼蓝排斥试验以及在灌注过程中标记亮氨酸线性掺入三氯乙酸沉淀蛋白来判断。细胞分离后,从细胞中提取总磷脂或磷脂酰胆碱。结果表明,无论肺脏接受的是天然大鼠表面活性剂还是牛源性表面活性剂制剂,表面活性剂磷脂酰胆碱标记物掺入细胞磷脂酰胆碱的量都与时间呈线性关系。Ⅱ型细胞从肺泡摄取天然表面活性剂的量大约是摄取牛源性物质的三倍。给予0.5μmol表面活性剂磷脂或2.0μmol的结果相似,只是给予2.0μmol天然大鼠表面活性剂会使灌注1小时时摄取量达到饱和。无论以细胞DNA还是细胞磷脂酰胆碱的比活性为基础来表示结果,细胞相关磷脂酰胆碱都会增加。在3小时时,大部分给予的表面活性剂标记物仍与灌洗液相关,而其余部分与分离的细胞或细胞制备过程中的碎片相关。在灌注培养基中几乎检测不到标记物。(摘要截短至250字)

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