花青素Cyanidin-3-O-glucoside对脂多糖和白介素-4刺激的小鼠骨髓巨噬细胞极化的影响

Effect of anthocyanin Cyanidin-3-O-glucoside on bone marrow derived macrophages polarization induced by lipopolysaccharide or IL-4 in mice

目的:探讨树莓花青素矢车菊素-3-O-葡萄糖苷(C3G)对小鼠骨髓巨噬细胞(BMDM)炎性细胞因子表达和生成的影响,进而了解C3G调控BMDM极化的机制。方法:取小鼠骨髓细胞,用100 ng/ml巨噬细胞集落刺激因子(M-CSF)诱导为BMDM。将诱导成功的BMDM分为两组,一组BMDM经C3G预处理12 h,再与1 μg/ml脂多糖(LPS)共处理12 h;另一组BMDM经C3G和20 ng/ml IL-4共处理24 h。采用MTT法检测不同浓度C3G对BMDM细胞活性的影响,qRT-PCR法和ELISA法分别检测BMDM促炎细胞介质IL-6、IL-1β、TNF-α、MCP-1、iNOS和抑炎细胞介质IL-10、Arg-1的mRNA水平和蛋白水平。结果: MTT检测结果显示1、5、10、15、20、25和30 μg/ml的C3G对BMDM细胞活力无明显可见的影响,qRT-PCR和ELISA分析结果显示1 μg/ml LPS显著上调BMDM促炎细胞介质的表达,20 ng/ml IL-4显著上调BMDM抑炎细胞介质的表达;5、10和20 μg/ml C3G处理既可明显降低LPS刺激的BMDM促炎细胞介质表达,也可增强IL-4刺激的BMDM抑炎细胞介质表达。结论: C3G通过下调巨噬细胞促炎细胞介质表达和上调巨噬细胞抑炎细胞介质表达,调控BMDM极化,并影响巨噬细胞的炎性反应。

Objective: To investigate the anti-inflammatory effect of Cyanidin-3-O-glucoside (C3G),a major anthocyanin found in raspberry,on bone marrow-derived macrophages (BMDMs) in rodents.To understand the mechanism of C3G modulating BMDM polarization,the mRNA level and production of inflammatory mediators in BMDMs were analyzed. Methods: The mice bone marrow cells were harvested and, in vitro ,induced into BMDMs with 100 ng/ml M-CSF.These BMDMs were divided into two groups.The one group was pretreated with C3G for 12 h,and was co-cultured with C3G and 1 μg/ml lipopolysaccharide (LPS) for 12 h.The another group was co-cultured with C3G and 20 ng/ml interleukin 4 (IL-4) for 24 h.The BMDM cell viability was detected with MTT assay.Both mRNA level and production of inflammatory mediators,IL-6,IL-1β,TNF-α,MCP-1,iNOS,IL-10 and Arg-1,in BMDM cells,were analyzed with qRT-PCR and ELISA,respectively. Results: There were no cytotoxic effects of 1,5,10,15,20,25 and 30 μg/ml C3G on cell viability of BMDMs by MTT assay.LPS up-regulates the expression and production of proinflammatory mediators,IL-6,IL-1β,TNF-α,MCP-1 and iNOS.IL-4 up-regulates the expression and production of anti-inflammatory mediators,IL-10 and Arg-1.After 5,10 and 20 μg/ml C3G treatment for 12-24 h,the expression and production of proinflammatory mediators were suppressed in LPS-stimulated BMDMs,but the ones of anti-inflammatory mediators were enhanced in IL-4-stimulated BMDMs,in dose-dependent manner. Conclusion: C3G can modulate the BMDMs polarization through inhibiting the expression of proinflammatory mediators and enhancing the expression of anti-inflammatory mediators,and involve in inflammatory response in macrophages.