Phospholipid metabolism of hypothermically stored rat hepatocytes.
Phospholipid metabolism of hypothermically stored rat hepatocytes.
复制标题
低温保存的大鼠肝细胞的磷脂代谢。
DOI：
10.1002/hep.510300531
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发表时间：
1999
期刊：
影响因子：
--
通讯作者：
Southard,JH
中科院分区：
文献类型：
--
作者：
Kim,JS;Southard,JH
将分离的大鼠肝细胞悬浮并储存在Liebovitz-15培养基（37 ℃或4 ℃）或含有[3 H]花生四烯酸（AA）的威斯康星州大学（UW）溶液（4 ℃）中.在不同的时间，通过薄层色谱法分离膜磷脂。AA标记的磷脂在4 ℃和37 ℃下相似.对双标记细胞中[3 H] AA和[14 C]甘油掺入磷脂酸或其他磷脂的比例的分析表明，脱酰/再酰化循环是低温下AA掺入的主要途径。这一点得到了支持，表明阻断磷脂酶A2（PLA 2）活性的三氟拉嗪抑制AA掺入磷脂。通过测定AA的释放来测定PLA 2活性，在48小时的冷藏期间是缓慢的，但是当通过抑制线粒体和糖酵解而耗尽ATP时，PLA 2活性显著增加。在整个大鼠肝脏中，在48小时储存期间没有显著的磷脂损失（总磷脂[μmol磷/L/mg]：0.197±. 001），除非使用能量阻滞剂（0.155±. 005）或通过禁食消耗糖原（0.167± 0.005）。001在48小时）。这项研究表明，净PLA 2刺激的磷脂水解被视为只有当ATP耗尽，其产生从厌氧糖酵解抑制。因此，当肝脏在最佳条件下获得时，磷脂的PLA 2水解不是冷藏期间肝细胞损伤的重要原因。然而，在冷藏过程中，影响ATP生成的条件可能会改变PLA 2，导致膜损伤。
Isolated rat hepatocytes were suspended and stored in either Liebovitz-15 medium (37 C or 4 C) or University of Wisconsin (UW) solution (4 C) containing [3 H] arachidonic acid (AA). At varying times, membrane phospholipids were separated by thin layer chromatography. AA labeled phospholipids similarly at both 4 C and 37 C. Analysis of the ratios of [3 H] AA and [14 C] glycerol incorporated into phosphatidic acid or other phospholipids in dual-labeled cells indicated that the deacylation/reacylation cycle was the major route of AA incorporation at hypothermia. This was supported by showing that blocking phospholipase A 2 (PLA 2) activity by trifluoperazine suppressed AA incorporation into phospholipids. PLA 2 activity, measured by determining the release of AA, was slow during 48-hour cold storage, but increased significantly when ATP was depleted by inhibition of mitochondria and glycolysis. In the whole rat liver, there was no significant loss of phospholipids during 48-hour storage (total phospholipids [μmol phosphorus/L/mg]: 0.197±. 001 at 0 hours) unless energy blockers were used (0.155±. 005 at 48 hours) or glycogen depleted by fasting the rat (0.167±. 001 at 48 hours). This study shows that a net PLA 2 stimulated hydrolysis of phospholipids is seen only when ATP is depleted and its generation from anaerobic glycolysis inhibited. Thus, PLA 2 hydrolysis of phospholipids is not a significant cause of liver cell injury during cold storage when livers are obtained in optimal condition. However, conditions affecting the generation of ATP during cold storage could alter PLA 2 leading to membrane damage.
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