Phospholipid-deacylating activities of the mouse peritoneal macrophages during phagocytosis
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概要
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In the macrophages (Mφ) 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 Mφ hydrolyzed PE more than PC.Exogenously added Ca2+ did not increase the enzymatic activities.A comparison was made of three kinds of the Mφ-phospholipid deacylating activities at pH8, after challenging the Mφ with Mycobacterium lepraemurium, Escherichia coli, zymosan, or latex beads for 17 hours at 37°C. The bacteria used to the phagocytosis were autoclaved.When the Mφ were challenged with M. lepraemurium, the phospholipase A1, A2 and lyso-phospholipase 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 200μg of zymosan. The increase in phospholipase A2 activity of the Mφ pretreated with the bacteria or zymosan seems to result in an increase in the hydrolysis of arachidonic acid from the Mφ-phospholipids to synthesize its inflammatory oxygenated metabolites.The lysophospholipase activity was not stimulated by the substances used to challenge the Mφ, except for M. lepraemurium. No significant increase in three kinds of phospholipid-deacylating activities was observed after challenging the Mφ with latex beads.It was considered from the above results that the Mφ-phospholipid-deacylating activities at pH8 might be affected by sort of the ingested substances.
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