目的检测RANTES对肥大细胞IL-10和IL-12分泌的影响和可能的信号转导通路。方法 P815肥大细胞培养、激发后收集不同时间点的细胞和上清液,用酶联免疫吸附试验(ELISA)检测上清中IL-10和IL-12的水平,用细胞激发信号ELISA(CASE)方法检测丝/苏氨酸蛋白激酶(Akt),细胞外信号调节激酶(ERK),信号转导子和转录激活子(STAT)3和p38丝裂原活化蛋白激酶信号转导通路蛋白磷酸化情况。结果 RANTES能够促进肥大细胞P815分泌IL-10而对IL-12的分泌无明显影响。LY294002能够阻断RANTES引起的肥大细胞IL-10分泌并抑制RANTES引起的Akt磷酸化。结论 RANTES刺激肥大细胞P815分泌IL-10可能是通过激活Akt信号转导通路实现的。
This study is aimed to investigate the potential influence of RANTES on IL-10 and IL-12 release from P815 mast cells as well as the potential signal transduction pathways correspondingly.After P815 mast cells were challenged with different concentrations of RANTES,the supernatant of challenged cells was analyzed by enzyme-linked immunosorbent assay(ELISA) to detect the release of IL-10 and IL-12,and the challenged cells were treated and analyzed by cellular activation of signal ELISA(CASE) to detect phosphorylation of serine/threonine protein kinase(Akt),extracellular signal-regulated kinase(ERK),p38 mitogen-activated protein kinase(MAPK),as well as signal transducer and activators of transcription 3(STAT3).We found that LY294002 not only blocked IL-10 release but also inhibited the RANTES-induced phosphorylation of Akt in P815 mast cells.All this results indicate that RANTES-stimulated IL-10 release may relate to activation of Akt signalling pathways in P815 mast cells.