Matsuki G, Kashiwabara Y, Nakagawa H
Nihon Rai Gakkai Zasshi. 1989 Oct-Dec;58(4):259-69. doi: 10.5025/hansen1977.58.259.
In the macrophages (M phi) obtained from mouse peritoneal exudates, five kinds of phospholipid-deacylating activities were detected using phosphatidylethanolamine (PE) and phosphatidylcholine (PC) labeled with [1-14C]oleic acid either in 1- or 2- position and 1- [1-14C]oleoyl-lysoPE, as substrates. Two types of phospholipipase A1 with pH optima of 4 to 6 and 8, respectively, and two types of phospholipase A2 activities with pH optima of 4 to 5 and 8, respectively, were identified. A detected lysophospholipase activity exhibited a broad pH optimum between 4 and 8. Both types of the phospholipase A1 and A2 of M phi hydrolyzed PE more than PC. Exogenously added Ca2+ did not increase the enzymatic activities. A comparison was made of three kinds of the M phi-phospholipid deacylating activities at pH8, after challenging the M phi with Mycobacterium lepraemurium, Escherichia coli, zymosan, or latex beads for 17 hours at 37 degrees C. The bacteria used to the phagocytosis were autoclaved. When the M phi were challenged with M. lepraemurium, the phospholipase A1, A2 and lysophospholipase activities were stimulated by about 160%, 150% and 140%, respectively. However, when challenged with E. coli, the phospholipase A1 activity remarkably decreased by about a third, although the phospholipase A2 activity was stimulated by about 150% that is similar to the challenge with M. lepraemurium. An inflammatory substance, zymosan seemed an effective inducer of the phospholipase A2, the enzymatic activity was remarkably stimulated by 260%, when challenged with 200 micrograms of zymosan. The increase in phospholipase A2 activity of the M phi pretreated with the bacteria or zymosan seems to result in an increase in the hydrolysis of arachidonic acid from the M phi-phospholipids to synthesize its inflammatory oxygenated metabolites. The lysophospholipase activity was not stimulated by the substances used to challenge the M phi, except for M. lepraemurium. No significant increase in three kinds of phospholipid-deacylating activities was observed after challenging the M phi with latex beads. It was considered from the above results that the M phi-phospholipid-deacylating activities at pH8 might be affected by sort of the ingested substances.
在从小鼠腹腔渗出液中获得的巨噬细胞(M phi)中,以[1-14C]油酸标记于1位或2位的磷脂酰乙醇胺(PE)和磷脂酰胆碱(PC)以及1-[1-14C]油酰-溶血PE作为底物,检测到了五种磷脂脱酰基活性。鉴定出两种pH最佳值分别为4至6和8的磷脂酶A1,以及两种pH最佳值分别为4至5和8的磷脂酶A2活性。检测到的溶血磷脂酶活性在4至8之间呈现出较宽的pH最佳值。M phi的两种类型的磷脂酶A1和A2对PE的水解作用均大于对PC的水解作用。外源添加的Ca2+并未增加酶活性。在37℃下用鼠麻风杆菌、大肠杆菌、酵母聚糖或乳胶珠对M phi进行17小时刺激后,比较了pH8时的三种M phi磷脂脱酰基活性。用于吞噬的细菌经过高压灭菌。当用鼠麻风杆菌刺激M phi时,磷脂酶A1、A2和溶血磷脂酶活性分别被刺激约至160%、150%和140%。然而,当用大肠杆菌刺激时,磷脂酶A1活性显著下降约三分之一,尽管磷脂酶A2活性被刺激约150%,这与用鼠麻风杆菌刺激的情况相似。一种炎性物质酵母聚糖似乎是磷脂酶A2的有效诱导剂,当用200微克酵母聚糖刺激时,酶活性被显著刺激至260%。用细菌或酵母聚糖预处理的M phi的磷脂酶A2活性增加似乎导致从M phi磷脂中水解花生四烯酸以合成其炎性氧化代谢产物的增加。除了鼠麻风杆菌外,用于刺激M phi的物质均未刺激溶血磷脂酶活性。在用乳胶珠刺激M phi后,未观察到三种磷脂脱酰基活性有显著增加。从上述结果认为,pH8时的M phi磷脂脱酰基活性可能受摄入物质种类的影响。