激活素A对小鼠中性粒细胞活性的调控作用

摘要

目的：通过检测中性粒细胞激活素受体表达以及细胞活性，阐述激活素A调控中性粒细胞的作用。方法：分离小鼠腹腔中性粒细胞；通过免疫荧光细胞化学染色及流式细胞术检测中性粒细胞激活素ⅡA型受体（ ActRⅡA）表达；激活素A刺激后，Western blot检测中性粒细胞Smad3表达；检测呼吸爆发、NO分泌及吞噬能力的变化以确定中性粒细胞活性。结果：分离的小鼠腹腔中性粒细胞纯度大于90％；免疫荧光细胞化学染色显示中性粒细胞可以表达ActRⅡA，流式细胞术检测结果显示Gr－1与ActRⅡA双阳性细胞约41．1％；激活素A刺激后，中性粒细胞p－Smad3表达上调，ROS及O2－产生升高（ P＜0．05），NO分泌增加（P＜0．01），流式细胞术检测结果显示激活素A还可以明显促进中性粒细胞对荧光微球的吞噬（P＜0．01）。结论：中性粒细胞可以表达激活素受体及其信号传导蛋白，激活素A对中性粒细胞活化及功能具有重要的调节作用。%Objective:To study the role of activin A in regulation of neutrophil function by detecting activin receptor expression and cellular activities.Methods:Peritoneal neutrophils were isolated in mouse.After the neutrophils were stimulated with activin A,the expression of ActRⅡA on neutrophils was examined by immunofluorescence and flow cytometry.Expression of Smad3 in neutrophils was analyzed by Western blot.Assays of neutrophils function were performed by detecting respiratory burst, production of NO and phagocytosis.Results:The isolated cells were composed of more than 90% peritoneal neutrophils.ActRⅡA was expressed on mouse neutrophils and Gr-1/ActRⅡA double-positive cells were 41.1%.Activin A promoted Smad3 phosphorylation in neutrophils,increased the production of ROS and O2-(P<0.05),enhanced secretion of NO and phagocytosis of mouse neutrophils(P<0.01),and promoted fluorescent microsphere phagocytosis of neutrophils by flow cytometry ( P<0.01 ) .Conclusion: Activin receptor and activin signaling protein were expressed on mouse neutrophils,activin A might play an important regulatory role in activation and function of neutrophils.