Birkle D L, Ellis E F
Neurochem Res. 1983 Mar;8(3):319-32. doi: 10.1007/BF00965722.
Arachidonic acid (AA) incorporation into phospholipids and cyclooxygenase and lipoxygenase mediated metabolism of arachidonic acid were studied in homogenized and intact Neuro-2A cells. When 3H8-AA was added to homogenized cells and incubated 20 minutes, 39% of the label was converted to prostaglandins (PGs), 10% to hydroxy-eicosatetraenoic acid (HETE) and 26% was incorporated into phospholipids. PGE2 and PGF2a were the major PGs produced. Synthesis of PGs was blocked by 10 microM indomethacin and synthesis of PGs and HETE was blocked by 10 microM eicosatetraynoic acid (ETYA). The cell homogenate produced the 13,14-dihydro-15-keto metabolites of PGE2 and PGF2a from 3H8-AA and also converted exogenous 3H7-PGE2 and 3H8-PGF2a to metabolites. When intact cells were labeled for 24 hours with 14C1-AA and the cells and media then analyzed, 75% of the radioactivity was incorporated into cellular phospholipids, 0.8% was converted to PGs and metabolites and 0.7% converted to HETE. Cells prelabeled for 24 hours were washed and incubated for 30 minutes in fatty acid free media. There was a 23% release of AA from phospholipids. One-fifth of the released AA was converted to HETE. PG synthesis in the intact resting cells was low. In summary, the Neuro-2A cell provides a good model system for studying arachidonic acid metabolism and incorporation into phospholipids in cells of neuronal origin.
在匀浆的和完整的Neuro-2A细胞中研究了花生四烯酸（AA）掺入磷脂以及环氧化酶和脂氧化酶介导的花生四烯酸代谢。当将3H8-AA添加到匀浆细胞中并孵育20分钟时，39%的标记物转化为前列腺素（PGs），10%转化为羟基二十碳四烯酸（HETE），26%掺入磷脂中。PGE2和PGF2a是产生的主要PGs。PGs的合成被10 microM吲哚美辛阻断，PGs和HETE的合成被10 microM二十碳四烯酸（ETYA）阻断。细胞匀浆从3H8-AA产生了PGE2和PGF2a的13,14-二氢-15-酮代谢物，并且还将外源性3H7-PGE2和3H8-PGF2a转化为代谢物。当用14C1-AA对完整细胞进行24小时标记，然后分析细胞和培养基时，75%的放射性掺入细胞磷脂中，0.8%转化为PGs和代谢物，0.7%转化为HETE。预先标记24小时的细胞经洗涤后在无脂肪酸培养基中孵育30分钟。有23%的AA从磷脂中释放出来。释放的AA中有五分之一转化为HETE。完整静息细胞中的PG合成较低。总之，Neuro-2A细胞为研究花生四烯酸代谢以及其在神经元来源细胞中掺入磷脂提供了一个良好的模型系统。
