青藤碱对LPS、IL-4诱导的小鼠RAW264.7巨噬细胞极化的影响

Effect of sinomenine on mouse RAW264. 7 macrophage cells line polarization induced by LPS or IL-4

目的:探讨青藤碱(Sinomenine,SIN)对脂多糖(LPS)以及白细胞介素4(IL-4)诱导的RAW264.7细胞向M1、M2型极化的影响。方法:以LPS刺激RAW264.7细胞诱导M1型极化,IL-4刺激RAW264.7细胞诱导M2型极化;青藤碱作用于LPS或IL-4诱导的巨噬细胞后:用酶联免疫法(ELISA)检测不同诱导状态下RAW264.7细胞TNF-α和IL-10的分泌量;荧光定量PCR检测与巨噬细胞极化相关的精氨酸酶-1(Arg-1)、一氧化氮合酶(i NOS)、细胞因子信号转导抑制蛋白-2(SOCS2)和细胞因子信号转导抑制蛋白-3(SOCS3)的mRNA表达水平。结果:青藤碱能抑制LPS诱导下细胞TNF-α的分泌量,抑制细胞i NOS和SOCS3的mRNA表达水平的升高。青藤碱能抑制IL-4诱导下细胞IL-10的分泌量和Arg1的mRNA表达水平的升高,对IL-4诱导下细胞SOCS2的mRNA表达水平的升高没有明显影响。结论:青藤碱对LPS诱导下巨噬细胞向M1型极化具有抑制作用;对IL-4诱导下巨噬细胞向M2型极化具有抑制作用。青藤碱对M1/M2亚型的失衡具有调节作用,有利于维持其动态平衡。

Objective： To investigate sinomenine（ Sinomenine,SIN） effect on RAW264. 7 cells polarization to M1 or M2 phenotype induced by lipopolysaccharide（ LPS） or interleukin-4（ IL-4）. Methods： RAW264. 7 cells were induced to polarize to M1 by LPS,and to M2 by IL-4. Sinomenine effects on LPS or IL-4 induced macrophages： TNF-α and IL-10 secretion induced by different condition were detected by Enzyme linked immunosorbent assay（ ELISA）; The expression level of mRNA of Arginase1（ Arg-1）,Nitric oxide synthase（ i NOS）,suppressor of cytokine signaling protein-2（ SOCS2） and suppressor of cytokine signaling protein-3（ SOCS3） of M1 / M2 phenotypes were detected by real time PCR respectively. Results： Sinomenine inhibited the increase of TNF- α secretion,i NOS and SOCS3 mRNA expression level induced by LPS. Sinomenine inhibited the increase of IL-10 secretion and Arg-1 mRNA expression level induced by IL-4,but SOCS2 mRNA expression level was not affected by Sinomenine. Conclusion： Sinomenine can inhibite the macrophage polarization to M1 and M2 induced by LPS and IL-4. Sinomenine plays a regulatory role on imbalance of M1 / M2,and is conducive to maintain the dynamic balance.