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目的检测白细胞介素(IL)12对肥大细胞介质分泌的影响并探讨其可能的信号转导通路。方法肥大细胞P815培养、激发后收集细胞和上清液,用ELISA法检测上清液中组胺、IL-4和IL-6的水平,用细胞激发信号ELISA(CASE)方法检测蛋白激酶B(AKT)、细胞外信号调节的蛋白激酶(ERK)、信号转导子和转录激活子(STAT)3和p38信号转导通路蛋白磷酸化情况。结果IL-12以浓度依赖的方式促进肥大细胞P815分泌IL-4,但对肥大细胞IL-6的分泌和组胺释放无明显影响。PD98059,U0126和LY294002阻断IL-12引起的肥大细胞IL-4分泌,并抑制IL-12引起的ERK和AKT磷酸化。结论IL-12刺激肥大细胞P815分泌IL-4很可能是通过激活AKT和ERK信号转导通路实现的。
Objective To investigate the effect of interleukin (IL) 12 on the secretion of mast cells and explore the possible signal transduction pathways. Methods M8 cells were cultured and stimulated with P815. The levels of histamine, IL-4 and IL-6 in the supernatants were detected by ELISA. Protein kinase B (CASE) AKT), extracellular signal-regulated protein kinase (ERK), signal transducers and activators of transcription (STAT) 3 and phosphorylation of the p38 signaling pathway. Results IL-12 promoted the secretion of IL-4 by mast cell P815 in a concentration-dependent manner, but had no significant effect on the secretion of IL-6 and histamine release in mast cells. PD98059, U0126 and LY294002 blocked IL-12-induced mast cell IL-4 secretion and inhibited IL-12-induced ERK and AKT phosphorylation. Conclusion IL-12 stimulates the secretion of IL-4 by P815 in mast cells probably through the activation of AKT and ERK signal transduction pathway.