脂氧素A4抑制过度激活状态中性粒细胞分泌促炎细胞因子的研究

Inhibitory effects of LXA4 on hyperirritable PMN to secrete proinflammatory cytokines in vitro

目的观察脂氧素A4对激活的中性粒细胞（PMN）分泌促炎细胞因子的影响。方法（1）采用5mg／L脂多糖（LPS）刺激后行PMN培养，24h后测定其白细胞介素（IL）-1β、肿瘤坏死因子（TNF）-α浓度；（2）采用1、10、100、1000ug／L的LXA4预处理，再加入5mg／LLPS共培养24h后，测定其IL-1β、TNF-α浓度；（3）采用100ug／LLX A4预处理，再加入LPS共培养，按时序测定其IL-1β、TNF-α浓度。结果（1）第一阶段：LPS组IL-1β、TNF-α浓度约为对照组的6、4倍（P〈0．01）；（2）第二阶段：不同浓度LXA4组相对LPS组的IL-1β、TNF—α浓度均有显著下降（P〈0．01），LXA4浓度越高，其上清液细胞因子浓度越低，呈剂量依赖性（P〈0．01或P〈0．05）。（3）第三阶段：培养时间越长，LPS组IL-1β、TNF-α浓度越高，16h达高峰（P〈0．01）；在6、8、16、24h时点，LXA4组与LPS组比较，TNF—α、IL-1β浓度显著下降（P〈0．05）。结论LXA4对LPS激活的PMN分泌IL-1β、TNF-α具有抑制作用，且呈剂量依赖性，LXA4峰值浓度为100o,g／L；其中TNF—α的抑制似乎早于IL-1β。

Objective To study the role of LXA4 to inhibit activated polymorphonuclear neutrophils （PMN） to secrete proinflammatory cytokines. Methods （1） Cultured PMN were routinely incubated with LPS （5 mg/L） for 24 h,and the contents of IL-1β,and TNF-α in culture fluid were determined. （2） PMN were pretreated with LXAd at different concentrations （1,10,100,and 1000 uL） ,then incubated with LPS （5 mg/L） for 24 h. The contents of IL-1β ,and TNF-α in culture fluid were determined. （3） PMN were pretreated by LXA4 （ 100 ug/L） ,then incubated with LPS （5 mg/L） for 24 h. The contents of IL-1β, and TNF-α in culture fluid were determined chronologically. Results （ 1 ） In phase 1 ： The concents of IL-1β and TNF-α in culture fluid of LPS group were 6 and 4 times of those in control group （ P 〈 0.01 ） ; （ 2 ） In phase 2 ： The contents of IL-1β, and TNF-α in LXA4-pretreated groups were decreased significantly as compared with LPS group （P 〈 0. 01 ）. With the increases in LXA4 concentrations, the contents of IL-1β, and TNF-α were also decreased significantly （ P 〈 0.01 or P 〈 0.05 ） ; （3） In phase 3 ： With the extension of time, the contents of IL-1β, and TNF-α in LPS group were increased gradually, and reached the peak at 16 h. On the 6th, 8th, 16th, and 24th h, the contents of TNF-α, and IL-1β in LXA4-treated group were decreased significantly as compared with LPS group （ P 〈 0.05 ）. Conclusion LXA4 can inhibit the production of IL-1β and TNF-α secreted by PMN which were activated by LPS in a dose-dependent manner. The peak concentrations of LXA4 was 100 ug/L. The inhibitory effect of LXA4 on TNF-α may be anterior to IL-1β.