川续断皂苷Ⅵ对M1/M2型巨噬细胞极化的调节作用及机制

Effect and mechanism of AsperosaponinⅥon polarization of M1/M2 macrophages

目的:研究川续断皂苷Ⅵ对M1/M2型巨噬细胞极化的调节作用及机制。方法:MTT法检测川续断皂苷Ⅵ对RAW264.7细胞活力的影响。ELISA检测脂多糖(LPS)诱导状态下RAW264.7细胞上清中TNF-α和IL-6的分泌量;Griess法检测LPS诱导状态下RAW264.7细胞上清中一氧化氮(NO)含量;荧光定量PCR检测TNF-α、IL-6、精氨酸酶-1(Arg-1)、血红素加氧酶1(HO-1)和细胞因子信号转导抑制蛋白-2(SOCS2)基因表达水平。Western blot检测一氧化氮合酶(iNOS)和p-p65蛋白表达。结果:在LPS诱导的RAW264.7细胞中,川续断皂苷Ⅵ抑制TNF-α、IL-6、iNOS和p-p65蛋白或基因表达水平,同时增加HO-1基因表达。川续断皂苷Ⅵ能抑制LPS诱导下RAW264.7细胞分泌的NO。川续断皂苷Ⅵ增加IL-4诱导下M2型巨噬细胞标志物Arg1和SOCS2基因表达。结论:川续断皂苷Ⅵ可以抑制RAW264.7巨噬细胞向M1型极化,同时促进其向M2型极化,可通过调节M1/M2型巨噬细胞极化发挥其抗炎免疫调节作用。

Objective:To study the regulation and mechanism of AsperosaponinⅥon polarization of M1/M2 macrophages.Methods:MTT assay was used to detect the effects of AsperosaponinⅥon RAW264.7 cell viability.The levels of TNF-αand IL-6 in supernatant of RAW264.7 cells induced by lipopolysaccharide(LPS)were determined by ELISA.The content of nitric oxide(NO)in supernatant of RAW264.7 cells induced by LPS was determined by Griess method.The gene expression levels of TNF-α,IL-6,argi⁃nase-1(Arg-1),heme oxygenase-1(HO-1)and suppressor of cytokine signaling protein-2(SOCS2)were detected by fluorescence quantitative PCR.Western blot was used to detect the expression levels of iNOS and p-p65 protein.Results:In LPS induced RAW264.7 cells,AsperosaponinⅥinhibited protein or gene expression levels of TNF-α,IL-6,iNOS and p-p65,and increased HO-1 gene expression.AsperosaponinⅥinhibited NO secretion in RAW264.7 cells induced by LPS.AsperosaponinⅥincreased the gene expression levels of M2 macrophage markers Arg1 and SOCS2 induced by IL-4.Conclusion:AsperosaponinⅥinhibited RAW264.7 macrophage polarization to M1 type and promote it polarization to M2 type,which can play its anti-inflammatory and immunomodulato⁃ry role by regulating M1/M2 macrophage polarization.