CB-839通过抑制谷氨酰胺代谢对巨噬细胞向M1型极化的影响

CB-839 resets macrophage polarization towards M1 phenotype by inhibiting glutamine metabolism

目的:研究谷氨酰胺酶抑制剂CB-839对巨噬细胞极化分型的影响。方法:准备小鼠骨髓来源巨噬细胞(bone marrow-derived macrophages,BMDM)和RAW264.7细胞,分为对照组和CB-839处理组。以CCK8检测巨噬细胞的增殖,用ELISA法检测M1型巨噬细胞相关因子肿瘤坏死因子α(TNF-α)和白介素1β(IL-1β)以及M2型巨噬细胞相关因子肿瘤生长因子β(TGF-β)和IL-10,用流式细胞术检测M1型表面标志物CD86和M2型表面标志物CD206,用Western blot检测iNOS和Arg1蛋白表达水平。结果:CCK8结果显示CB-839并不影响BMDM和RAW264.7细胞的增殖,CB-839处理组的TNF-α和IL-1β的水平与CD86、iNOS的表达水平较对照组显著上升,TGF-β和IL-10的水平与CD206、Arg1的表达较对照组显著下降。结论:CB-839在不影响细胞增殖的前提下可以促进巨噬细胞向M1型极化。

Objective:To research the effect of glutaminase inhibitor CB-839 on the polarization of macrophages.Methods:Following treatment of bone marrow-derived macrophages(BMDM)and RAW264.7 cells with PBS or CB-839 for 48 h,the treated cells were divided into the the control group and the CB-839 group.The proliferation of cells was detected by CCK8 kit,the expression levels of tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β),TGF-βand IL-10 were detected by ELISA,the expression levels of CD86 and CD206 were detected by flow cytometry,and the expression levels of iNOS and Arg1 were detected by Western blot.Results:CCK8 detection showed that CB-839 almost had no effect on the proliferation of macrophages.The expression levels of TNF-α,IL-1β,CD86 and iNOS in the CB-839 group significantly elevated as compared with those of the control group,the expression levels of TGF-βand IL-10 as well as CD206 and Arg1 markedly down-regulated as compared with those of the control group.Conclusion:CB-839 could enhance macrophage polarization towards M1,based on the premise that it did not inhibit the proliferation of cells.