目的检测不同CD40L表达水平的脾细胞对巨噬细胞表型和功能的影响,探讨通过调控CD40L的表达改变巨噬细胞的类型。方法贴壁分离正常BALB/c小鼠的腹腔巨噬细胞。将腹腔巨噬细胞或经IL-4刺激的腹腔巨噬细胞,分别与体外培养2d后获得的CD40L^low脾细胞和Ionomycin与PMA诱导的CD40L^high脾细胞共孵育,24h后检测M1和M2型巨噬细胞相关基因的表达,并以硝酸还原酶法检测NO分泌水平。结果与CD40L^low脾细胞共孵育的巨噬细胞CCL22、Fizz1和Ym1表达水平较高,但只分泌较低水平的NO,而与CD40L^high脾细胞共孵育的巨噬细胞表达较高水平的CCL3并分泌高水平的NO,而且CD40L^high脾细胞可使M2型巨噬细胞表型相关基因明显下调。结论CD40L^high脾细胞使巨噬细胞倾向于M1型极化,并且能使M2型巨噬细胞向M1型转换。
Objective To investigate whether modulation of the expression level of CD40L on splenocytes could alter the type of macrophages. Methods In vitro 48-hour-cultured splenocytes were used as CD40L^low splenocytes; ionomycin and PMA-treated splenocytes were used as CD40L^high splenocytes. The BALB/c peritoneal macrophages were purified by adhering method, pre-treated with or without IL-4 was then incubated with CD40L^high or CD40L^low splenocytes. The expression levels of CCL3, CCL22, Fizzl and Yml were detected and the iNOS activity was determined to identify the type of macrophages. Results The macrophages co-cultured with CD40L^high splenocytes expressed higher level of CCL3 and secreted higher abundance of NO, while those co-cultured with CD40L^low splenocytes showed higher expression of CCL22, Fizzl and Yml, and produced less NO. Conclusions CD40L; splenocytes co-culture could induce M1 macrophage towards M1. polarization, and even convert the pre-existed M2