Hinder M
Department of Pharmacology, Boehringer Mannheim GmbH, Germany.
Arzneimittelforschung. 1998 Jan;48(1):77-81.
We investigated the effects of phospholipase A2 (PLA2) activation and inhibition on the chemotaxis of mixed human leukocytes (MHL) in a 48-well microchemotaxis chamber and on the production of leukotriene B4 (LTB4) by an enzyme immuno assay. LTB4 is the eicosanoid which is most likely responsible for induction of chemotaxis and its production can be used as an indicator of PLA2 stimulation. Formyl-methionyl-leucyl-phenylalanine (F-met-leu-phe, f-MLP), a well known inducer of PLA2 in our experiments stimulated dose-dependently the chemotaxis of MHL with a maximum at 10 nmol/l. In accordance with the chemotaxis assay the maximum LTB4-production was found at 10 nmol/l f-MLP. The preincubation of MHL with the PLA2 inhibitors mepacrine (1-100 mumol/l), manoalide (1-100 mumol/l) and BM 16.2115 (1,6-dioxadispiro[4.1.5.1]trideca-8,11-diene-2,10-dione, 0.5-50 mumol/l) dose-dependently reduced the chemotaxis of MHL induced by 10 nmol/l f-MLP. The inhibitory effect of the experimental compound BM 16.2115 was about 2 times stronger than those of the reference compounds mepacrine and manoalide. Thus inhibitors of PLA2 reduce chemotaxis of leukocytes probably by reducing production of LTB4. This mechanism could be of considerable interest for the development of anti-inflammatory drugs.
我们在48孔微量趋化性实验箱中研究了磷脂酶A2（PLA2）激活和抑制对人混合白细胞（MHL）趋化性的影响，并通过酶免疫测定法研究了其对白三烯B4（LTB4）产生的影响。LTB4是最有可能诱导趋化性的类二十烷酸，其产生可作为PLA2刺激的指标。在我们的实验中，甲酰甲硫氨酰亮氨酰苯丙氨酸（F-met-leu-phe，f-MLP）是一种众所周知的PLA2诱导剂，它以剂量依赖性方式刺激MHL的趋化性，在10 nmol/l时达到最大值。与趋化性测定结果一致，在10 nmol/l f-MLP时发现LTB4的产生量最大。用PLA2抑制剂米帕林（1 - 100 μmol/l）、 manoalide（1 - 100 μmol/l）和BM 16.2115（1,6 - 二氧杂双螺[4.1.5.1]十三碳 - 8,11 - 二烯 - 2,10 - 二酮，0.5 - 50 μmol/l）对MHL进行预孵育，可剂量依赖性地降低10 nmol/l f-MLP诱导的MHL趋化性。实验化合物BM 16.2115的抑制作用比参考化合物米帕林和manoalide强约2倍。因此，PLA2抑制剂可能通过减少LTB4的产生来降低白细胞的趋化性。这种机制对于抗炎药物的开发可能具有重要意义。
