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通过在非等渗培养基中培养对肾细胞系硫糖脂的适应性改变。

Adaptive changes in sulfoglycolipids of kidney cell lines by culture in anisosmotic media.

作者信息

Niimura Y, Ishizuka I

机构信息

Department of Biochemistry, Teikyo University School of Medicine, Tokyo, Japan.

出版信息

Biochim Biophys Acta. 1990 May 2;1052(2):248-54. doi: 10.1016/0167-4889(90)90218-3.

Abstract

(1) The effects of osmolarity environments on renal glycolipid composition were examined using established renal cell lines. The profile of glycosphingolipids of Madin-Darby canine kidney cells (MDCK) in culture with anisosmotic media showed that a hyposomotic medium reduced the concentration of GalCer I3-sulfate and LacCer II3-sulfate. (2) The concentrations of sulfoglycolipids were increased by maintaining the culture in a hyperosmotic media prepared by the addition of various sodium salts to the control isosmotic medium, while the contents of most of the neutral glycolipids were reduced. The hyperosomotic medium supplemented with nonelectrolytes, mannitol, sucrose or urea, also increased the concentration of sulfoglycolipids. (3) Both sulfoglycolipids were increased linearly with gradual increases of sodium chloride in the medium. Hyperosmolarity produced by the addition of a nonelectrolyte, mannitol, also increased the levels of sulfoglycolipids. In both series of media, the most prominent accumulation was observed in LacCer II3-sulfate. (4) The incorporation of radioactive sulfate into sulfoglycolipids was elevated in cells adapted to high NaCl or mannitol. The increase of the label was observed not only in MDCK but also in three other established cell lines of renal tubular origin, JTC-12, LLC-PK1 and MDBK. (5) It was established, using the culture system of homogeneous cell lines, that the mechanism of increasing the amount of sulfoglycolipids is independent of the integral regulatory mechanism of animals and resides in the renal epithelial cell itself. These results suggest that by culture in hyperosmotic media, the elevated level of intracellular cations stimulated the activity of GalCer and LacCer sulfotransferase, inducing the increased expression of sulfoglycolipids.

摘要

(1) 使用已建立的肾细胞系研究了渗透压环境对肾糖脂组成的影响。在等渗介质中培养的Madin-Darby犬肾细胞(MDCK)的糖鞘脂谱显示,低渗介质降低了硫酸神经酰胺I3和硫酸乳糖神经酰胺II3的浓度。(2) 通过在对照等渗介质中添加各种钠盐制备高渗介质来维持培养,可增加硫糖脂的浓度,而大多数中性糖脂的含量则降低。添加非电解质、甘露醇、蔗糖或尿素的高渗介质也会增加硫糖脂的浓度。(3) 随着培养基中氯化钠的逐渐增加,两种硫糖脂均呈线性增加。添加非电解质甘露醇产生的高渗也增加了硫糖脂的水平。在这两种系列的介质中,在硫酸乳糖神经酰胺II3中观察到最显著的积累。(4) 在适应高氯化钠或甘露醇的细胞中,放射性硫酸盐掺入硫糖脂的量增加。不仅在MDCK中观察到标记增加,在另外三种肾小管来源的已建立细胞系JTC-12、LLC-PK1和MDBK中也观察到了增加。(5) 使用同质细胞系培养系统确定,硫糖脂含量增加的机制独立于动物的整体调节机制,存在于肾上皮细胞本身。这些结果表明,通过在高渗介质中培养,细胞内阳离子水平的升高刺激了神经酰胺和乳糖神经酰胺磺基转移酶的活性,诱导了硫糖脂表达的增加。

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