Elsbach P, Levy S
J Clin Invest. 1968 Oct;47(10):2217-29. doi: 10.1172/JCI105907.
Incorporation in vitro of (32)P-labeled lysolecithin (LPC) or lysophosphatidylethanolamine (LPE) into respectively lecithin (PC) and phosphatidylethanolamine (PE) of rabbit granulocytes and alveolar macrophages was compared in the absence and in the presence of ingestible particles. Maximal synthesis of PC by intact cells occurred at added LPC concentrations of less than 0.05 mmole/liter, i.e., at levels found in plasma. Accumulation of PC-(32)P proceeded linearly for at least 30 min and varied directly with cell concentration. While per cell granulocytes and macrophages converted comparable amounts of medium LPC to cellular PC, per milligram of protein, the granulocytes were approximately four times more active than the much larger macrophages. After 30 min newly synthesized PC-(32)P represented as much as 5% of total granulocyte PC. For macrophages this fraction did not exceed 1%. Addition of polystyrene or zymosan particles to the cell suspension resulted in up to 3-fold stimulation of incorporation of LPC-(32)P or LPE-(32)P into their respective diacyl derivatives. This stimulation did not occur when the cells were homogenized. Breakdown of LPC to water-soluble products during phagocytosis of polystyrene particles was the same as at rest. By use of doubly labeled LPC, the mechanism of PC synthesis by the two cell types has been identified as direct acylation of medium LPC, both at rest and during engulfment. Evidence presented in the case of granulocytes suggests that the increased translocation of medium LPC-(32)P during phagocytosis and its conversion to PC represents net synthesis. The findings indicate that LPC, a normal constituent of plasma, can serve as substrate in PC synthesis by phagocytic cells. This mechanism of PC synthesis can account for appreciable addition of membrane PC, especially by granulocytes. It is proposed that stimulation of this pathway provides building blocks for increased membrane formation during phagocytosis.
在不存在和存在可摄取颗粒的情况下,比较了兔粒细胞和肺泡巨噬细胞分别将(32)P标记的溶血卵磷脂(LPC)或溶血磷脂酰乙醇胺(LPE)体外掺入卵磷脂(PC)和磷脂酰乙醇胺(PE)的情况。完整细胞合成PC的最大值出现在添加的LPC浓度低于0.05毫摩尔/升时,即血浆中发现的水平。PC-(32)P的积累至少持续30分钟呈线性,且与细胞浓度直接相关。虽然每细胞的粒细胞和巨噬细胞将相当量的培养基LPC转化为细胞内PC,但每毫克蛋白质,粒细胞的活性比大得多的巨噬细胞高约四倍。30分钟后,新合成的PC-(32)P占粒细胞总PC的比例高达5%。对于巨噬细胞,这一比例不超过1%。向细胞悬液中添加聚苯乙烯或酵母聚糖颗粒可使LPC-(32)P或LPE-(32)掺入其各自的二酰基衍生物的量增加多达3倍。当细胞匀浆时,这种刺激不会发生。聚苯乙烯颗粒吞噬过程中LPC分解为水溶性产物的情况与静止时相同。通过使用双标记的LPC,已确定两种细胞类型合成PC的机制是培养基LPC的直接酰化,无论是在静止还是吞噬过程中。粒细胞的情况表明,吞噬过程中培养基LPC-(32)P的转运增加及其向PC的转化代表净合成。这些发现表明,血浆中的正常成分LPC可作为吞噬细胞合成PC的底物。这种PC合成机制可解释膜PC的明显增加,尤其是粒细胞。有人提出,刺激这一途径可为吞噬过程中增加的膜形成提供构建块。