Altered eicosanoid production and phospholipid remodeling during cell culture
Altered eicosanoid production and phospholipid remodeling during cell culture
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DOI：
10.1194/jlr.m083030
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发表时间：
2018-03-01
影响因子：
6.5
通讯作者：
Murphy, Robert C.
中科院分区：
文献类型：
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作者：
Okuno, Toshiaki;Gijon, Miguel A.;Murphy, Robert C.
Lands循环对PUFAs的重塑是细胞中磷脂分子种类多样性的原因。还没有详细的研究磷脂分子种类的变化，作为一个结果，血清饥饿或消耗的PUFA，通常发生在组织培养。通过脂质组学策略检查了细胞培养对培养24、48或72 h的RAW 264.7细胞中磷脂分子种类的时间依赖性影响。然后刺激这些细胞产生花生四烯酸代谢物，这些代谢物来源于环氧合酶途径，血栓烷B-2、PGE（2）和PGD（2），以及5-脂氧合酶途径，白三烯（LT）B-4、LTC 4和5-HETE，这些代谢物随着培养时间的增加而减少。然而，20：3脂肪酸的5-脂氧合酶代谢产物LTB 3、全反式LTB 3、LTC 3和5-羟基二十碳三烯酸呈时间依赖性增加。含有磷脂的花生四烯酸的分子种类在细胞培养过程中急剧重塑，新的20：3酰基被填充到磷脂中以取代日益稀缺的花生四烯酸。此外，TNF的量。与培养24 h相比，培养72 h的细胞内PUFAs诱导的炎症反应明显增加，提示PUFAs的重构增强了炎症反应。这些研究支持Lands循环的快速运行，通过填充PUFA种类来维持细胞生长和活力;然而，没有足够的n-6脂肪酸，20：3 n-9积累，导致脂质介质生物合成和炎症反应改变。
The remodeling of PUFAs by the Lands cycle is responsible for the diversity of phospholipid molecular species found in cells. There have not been detailed studies of the alteration of phospholipid molecular species as a result of serum starvation or depletion of PUFAs that typically occurs during tissue culture. The time-dependent effect of cell culture on phospholipid molecular species in RAW 264.7 cells cultured for 24, 48, or 72 h was examined by lipidomic strategies. These cells were then stimulated to produce arachidonate metabolites derived from the cyclooxygenase pathway, thromboxane B-2, PGE(2), and PGD(2), and the 5-lipoxygenase pathway, leukotriene (LT) B-4, LTC4, and 5-HETE, which decreased with increasing time in culture. However, the 5-lipoxygenase metabolites of a 20:3 fatty acid, LTB3, all trans-LTB3, LTC3, and 5-hydroxyeicosatrienoic acid, timedependently increased. Molecular species of arachidonate containing phospholipids were drastically remodeled during cell culture, with a new 20:3 acyl group being populated into phospholipids to replace increasingly scarce arachidonate. In addition, the amount of TNF. induced by lipopolysaccharide stimulation was significantly increased in the cells cultured for 72 h compared with 24 h, suggesting that the remodeling of PUFAs enhanced inflammatory response. These studies supported the rapid operation of the Lands cycle to maintain cell growth and viability by populating PUFA species; however, without sufficient n-6 fatty acids, 20:3 n-9 accumulated, resulting in altered lipid mediator biosynthesis and inflammatory response.