Watanabe M, Ohuchi K, Sugidachi A, Hirasawa N, Hayashi Y, Tsurufuji S
Department of Biochemistry, Faculty of Pharmaceutical Sciences, Tohoku University, Miyagi, Japan.
Int Arch Allergy Appl Immunol. 1987;84(4):396-403. doi: 10.1159/000234456.
Using a model of allergic inflammation of air pouch type in rats, the platelet-activating factor (PAF) in the pouch fluid in the anaphylactic phase was analyzed. Anaphylactic reaction was induced by injecting an antigen (azobenzene-arsonate-conjugated acetyl bovine serum albumin) solution into a subcutaneous air pouch preformed on the dorsum of immunized rats. The pouch fluid was collected 30 min after the antigenic challenge, and chloroform extract was subjected to normal phase high-performance liquid chromatography to isolate two fractions, PAF and lyso-PAF. In the pouch fluid, however, there was little activity of PAF as examined by the aggregation of guinea pig platelets. The lyso-PAF fraction obtained was acetylated to PAF chemically with pyridine and acetic anhydride. This acetylated lyso-PAF fraction induced the aggregation of guinea pig platelets, which was inhibited dose-dependently by a PAF antagonist, CV-3988. The amount of lyso-PAF in the pouch fluid of the immunized group in the anaphylactic phase was significantly higher than that of the nonimmunized group. When (3H-)PAF was incubated with the supernatant fraction of the pouch fluid it was metabolized into lyso-PAF time-dependently. The significance of the higher level of lyso-PAF in the pouch fluid in the anaphylactic phase of allergic inflammation is discussed.
利用大鼠气囊型变应性炎症模型,分析了过敏反应期气囊液中的血小板活化因子(PAF)。通过向免疫大鼠背部预先形成的皮下气囊内注射抗原(偶氮苯 - 砷酸盐 - 共轭乙酰牛血清白蛋白)溶液诱导过敏反应。抗原攻击30分钟后收集气囊液,其氯仿提取物经正相高效液相色谱法分离出两个组分,即PAF和溶血PAF。然而,通过豚鼠血小板聚集检测发现,气囊液中PAF的活性很低。所得到的溶血PAF组分用吡啶和乙酸酐化学乙酰化转化为PAF。这种乙酰化的溶血PAF组分可诱导豚鼠血小板聚集,且该聚集被PAF拮抗剂CV - 3988剂量依赖性抑制。过敏反应期免疫组气囊液中溶血PAF的量显著高于未免疫组。当(3H -)PAF与气囊液的上清液组分一起孵育时,它会随时间代谢为溶血PAF。文中讨论了过敏炎症过敏反应期气囊液中溶血PAF水平升高的意义。