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目的研究磷脂酰胆碱特异性磷脂酶Ｄ（ＰＣＰＬＤ）在佛波酯诱导Ｕ９３７细胞的前列腺素Ｅ２（ＰＧＥ２）生物合成调节中的作用。方法二甲亚砜（ＤＭＳＯ）分化的Ｕ９３７细胞与［３Ｈ］烷基溶血ＰＣ一起孵育，对细胞内ＰＣ池进行同位素标记。［３Ｈ］烷基磷脂酸（ＰＡ）、［３Ｈ］烷基二脂酰甘油（ＤＡＧ）、［３Ｈ］烷基磷脂酰乙醇（ＰＥｔ）采用薄层层析法分离，并以液体闪烁法测定含量。ＰＥｔ的形成被认为ＰＬＤ活性的可信的特异指标。培养液中ＰＧＥ２含量应用酶免疫法测量。结果１２十四烷酸１３乙酸佛波酯（ＰＭＡ，１μｍｏｌ·Ｌ－１）兴奋Ｕ９３７细胞产生［３Ｈ］烷基ＰＡ与［３Ｈ］烷基ＤＡＧ。孵育合剂中包含乙醇（２５～１０ｍｌ·Ｌ－１）引起浓度依赖性地减少ＰＭＡ诱导的ＰＡ与ＤＡＧ积聚，以及增加ＰＥｔ积聚。时相研究显示ＰＭＡ诱导的ＰＡ积聚先于ＤＡＧ积聚。ＰＡ磷酸水解酶抑制剂普萘洛尔（２００μｍｏｌ·Ｌ－１）增加ＰＡ积聚的同时减少ＰＭＡ诱导的ＤＡＧ的生成。乙醇与普萘洛尔均不影响ＰＭＡ增加Ｕ９３７细胞的ＰＧＥ２生成。结论ＰＭＡ激活ＤＭＳＯ分化Ｕ９３７细胞的ＰＬＤ，但它不联系该细胞的ＰＧＥ２生物合成。说明ＰＭＡ?
Objective To study the role of phosphatidylcholine-specific phospholipase D (PCPLD) in the regulation of prostaglandin E2 (PGE2) biosynthesis induced by phorbol ester in U937 cells. Methods Dimethyl sulfoxide (DMSO) differentiated U-937 cells were incubated with [3H] -alkyl-hemolysis PC to label the intracellular PC pool. The contents of [3H] -alkylphosphatidic acid (PA), [3H] -alkyl-diallylglycerol (DAG) and [3H] -alkyl phosphatidylethanolamine (PEt) were determined by TLC. Liquid scintillation method for the determination of content. The formation of PEt is believed to be a credible and specific indicator of PLD activity. The culture medium PGE2 content should be measured by enzyme immunoassay. Results 12-tetradecanoic acid 13 acetic acid phorbol ester (PMA, 1μmol·L-1) excited U 937 cells produce []] alkyl PA and []] alkyl DAG. Incubation mixture containing ethanol (2 5 ~ 10ml · L-1) caused a concentration-dependent reduction of PMA-induced PA and DAG accumulation, and increased PEt accumulation. Time-phase studies showed that PMA induced PA accumulation prior to DAG accumulation. PA phosphorylase inhibitor propranolol (200 μmol·L -1) increased PA accumulation while reducing PMA-induced DAG production. Ethanol and propranolol did not affect PMA increased P9 production of U-937 cells. Conclusion PMA activates PLD of DMSO-differentiated U-937 cells, but it does not relate to the PGE2 biosynthesis of this cell. Explain PMA?