PI3K/AKT信号通路在SjCystatin诱导M2巨噬细胞分化过程中的影响

目的:进一步确定日本血吸虫半胱氨酸蛋白酶抑制剂(Sj Cystatin)诱导M2巨噬细胞分化的亚型及相关机制。方法:用ELISA、RT-q PCR或Western blot法测定IL-10、IL-12、巨噬细胞亚型表面标志物LIGHT(M2b)及Arg-1(M2a+M2c)的表达;用Western blot法测定AKT的磷酸化水平。结果:Sj Cystatin处理组在6 h、12 h和24h时IL-10表达量持续增加;处理12 h,LIGHT的mRNA和蛋白表达量增加但Arg-1的mRNA和蛋白表达量降低;AKT磷酸化水平增加。PI3K/AKT抑制剂处理组IL-10的释放量在12 h和24 h持续降低;24 h,LIGHT的mRNA和蛋白表达量降低但Arg-1的mRNA和蛋白表达量增加;AKT的磷酸化水平减少。结论:Sj Cystatin促进了活化的M2巨噬细胞分化为M2b亚型巨噬细胞,并且PI3K/AKT信号通路参与了这一过程。

Effects of PI3K/AKT signaling pathway on polarization of M2 macrophages induced by SjCystatin

AIM: To investigate the subtype of M2 macrophages induced by Schistosoma japonicum cystatin (SjCystatin) and to determine the mechanism underlying these effects.METHODS: The releases of IL-10 and IL-12, and the expression of macrophage subtype markers LIGHT (M2b) and Arg-1 (M2a+M2c) were assessed by ELISA, RT-qPCR and Western blot. The phosphorylation level of AKT was assessed by Western blot.RESULTS: SjCystatin promoted the continuous increase in IL-10 level at 6 h, 12 h and 24 h, and increased the amount of mRNA and protein expression of LIGHT, but down-regulated the amount of mRNA and protein expression of AKT. The addition of PI3K/AKT inhibitor reduced the release of IL-10 at 12 h and 24 h, reduced the mRNA and protein expression of LIGHT at 24 h, up-regulated the mRNA and protein expression of Arg-1 at 24 h, and decreased the phosphorylation level of AKT.CONCLUSION: SjCystatin promotes the differentiation of M2 macrophage to M2b macrophage subtype, and the PI3K/AKT signaling pathway is involved in this process.