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目的研究γ促分泌酶抑制剂(GSI)MW167阻断Notch信号后,呼吸道合胞病毒(RSV)毛细支气管炎患儿外周血单个核细胞(PBMC)中GATA结合蛋白3(GATA3)的变化。方法募集RSV毛细支气管炎30例,正常组25例。分离PBMC,分为正常对照组、RSV组、MW167处理组。ELISA检测培养细胞上清液中白细胞介素4(IL-4)水平,实时荧光定量PCR检测细胞中GATA3 mRNA水平,Western blot法检测细胞中Notch1胞内结构域(NICD)及GATA3蛋白的表达。结果与RSV组相比,MW167处理组IL-4水平降低,GATA3 mRNA、NICD及GATA3蛋白水平均显著降低。结论 GSI阻断Notch信号通路,可降低GATA3的水平。
Objective To investigate the changes of GATA binding protein 3 (GATA3) in peripheral blood mononuclear cells (PBMCs) of children with respiratory syncytial virus (RSV) bronchiolitis after G167 blockade of Notch signaling by gamma secretase inhibitor (GSI). Methods Thirty RSV bronchiolitis patients were recruited, and 25 patients in normal group. PBMCs were separated and divided into normal control group, RSV group and MW167 treatment group. The levels of IL-4 in supernatant of cultured cells were detected by ELISA. The level of GATA3 mRNA in cells was detected by real-time fluorescence quantitative PCR. The expression of Notch1 intracellular domain (NICD) and GATA3 protein was detected by Western blot. Results Compared with RSV group, the level of IL-4 in MW167 group was decreased and the levels of GATA3 mRNA, NICD and GATA3 protein were significantly decreased. Conclusion GSI blocks Notch signaling pathway and decreases GATA3 level.