CD30L基因表达对脂多糖刺激后小鼠腹腔CD11b^(+)细胞表型变化影响的研究

Study on CD30L affacting phenotypic changes of CD11b^(+)cells in mice stimulated by lipopolysaccharide

目的:建立小鼠无菌炎症模型,研究CD30L分子对脂多糖刺激CD11b^(+)细胞表型变化的影响。方法:①通过流式细胞术比较和分析注射巯基乙酸培养基(TGM)前后,小鼠腹腔CD11b^(+)细胞数量及比例变化;②使用流式细胞术检测在不同时间与浓度的脂多糖(LPS)刺激下,小鼠腹腔CD11b^(+)细胞表型变化的情况与CD30L分子表达的情况;③在LPS刺激CD11b^(+)细胞的小鼠无菌炎症模型中,分析和比较C57BL/6(WT)和Cd30l基因敲除小鼠(CD30LKO)腹腔CD11b^(+)细胞表型变化的差异。结果:小鼠腹腔注射TGM可募集大量CD11b^(+)细胞,注射TGM后WT小鼠腹腔CD11b^(+)细胞表面CD86、F4/80无明显变化,I-A/E、CD30L表达下降;随着LPS刺激时间和浓度的增加,经TGM注射后的WT小鼠腹腔CD11b^(+)细胞数量逐渐增加,CD11b^(+)细胞表面CD30L分子的表达逐渐增加,并且,CD86、F4/80、I-A/E的表达同样增加,同时,与WT小鼠相比,CD30LKO小鼠腹腔CD11b^(+)细胞表面CD86、I-A/E表达降低,CD206表达升高。结论:CD30L分子调控LPS刺激后的小鼠腹腔CD11b^(+)细胞表型的变化。

Objective:To establish a sterile inflammatory model in mice to study the effect of CD30L on the phenotypic changes of CD11b^(+)cells stimulated by lipopolysaccharide.Methods:①Flow cytometry was used to compare and analyze the changes in the number and proportion of peritoneal CD11b^(+)cells in mice before and after injection of Thioglycollate Medium(TGM).②Flow cytometry was used to detect the phenotype changes and CD30L molecular expression of peritoneal CD11b^(+)cells under the stimulus of lipopolysaccharide(LPS)at different concentrations and times.③Differences in phenotypic changes of peritoneal CD11b^(+)cells in C57BL/6(WT)and Cd30l knockout mouse(CD30LKO)were analyzed and compared in a mouse model of sterile inflammation stimulated by LPS.Results:Intraperitoneal injection of TGM in mice could recruit a large number of CD11b^(+)cells.CD86 and F4/80 on the surface of CD11b^(+)cells of WT mice showed no significant changes after intraperitoneal injection of TGM,while the expression of I-A/E and CD30L decreased.With the increase of time and concentration of LPS stimulation,after injected TGM,CD11b^(+)cells of WT mice increased gradually,the CD30L molecules on the surface of CD11b^(+)cells increased,and the expression of CD86,F4/80 and I-A/E also increased,meanwhile,compared with WT mice,while the expression of CD86 and I-A/E was decreased,the expression of CD206 was increased.Conclusion:CD30L regulates the phenotype of peritoneal CD11b^(+)cells in mice stimulated by LPS.