论文部分内容阅读
探究肉苁蓉低分子糖(LMSC)对RAW264.7小鼠巨噬细胞的激活作用靶点蛋白群及相关作用机制。该研究首先通过测定巨噬细胞吞噬活性以及NO释放量来评价LMSC对RAW264.7巨噬细胞的激活作用。实验结果显示,LMSC在0.25~2g·L-1给药浓度可显著提高RAW264.7细胞的吞噬活性及NO的释放量,提示具有巨噬细胞激活作用。通过构建LMSC键合的环氧活化琼脂糖固相微球作为亲和介质,捕获RAW264.7细胞裂解液中特异性结合靶标蛋白;对高分辨质谱鉴定获得的LMSC靶标蛋白群进行信号通路富集分析,探讨LMSC巨噬细胞激活作用相关机制。实验共获得Eef2等24个LMSC靶点蛋白,分别涉及Fcγ受体依赖的吞噬、TNF-αNF-κB信号通路、糖酵解/糖原异生以及柠檬酸(TCA)循环和呼吸电子运输过程等10条巨噬细胞激活相关信号通路。以上结果提示LMSC通过作用于多个靶点进而调节细胞吞噬、NF-κB信号通路以及糖代谢途径最终实现对RAW264.7巨噬细胞激活作用。
To investigate the protein targets of Cistanche deserticola (LMSC) -activated macrophages in RAW264.7 mice and its related mechanism. This study first evaluated the activation of LMSC on RAW264.7 macrophages by measuring macrophage phagocytic activity and NO release. The experimental results showed that the concentration of LMSC at 0.25 ~ 2g · L-1 could significantly increase the phagocytosis activity and NO release of RAW264.7 cells, suggesting the activation of macrophages. By constructing LMSC-bonded epoxy-activated agarose solid phase microspheres as the affinity medium, the target protein was captured in the lysis solution of RAW264.7 cells. The LMSC target protein population obtained by high resolution mass spectrometry was enriched in signal pathways Analysis, to explore LMSC macrophage activation related mechanisms. Twenty-four LMSC target proteins such as Eef2 were obtained, which involved Fcγ receptor-dependent phagocytosis, TNF-αNF-κB signaling, glycolysis / gluconeogenesis and citrate (TCA) cycling and respiratory electron transport Ten macrophages activate related signaling pathways. These results suggest that LMSC can activate RAW264.7 macrophages by regulating the phagocytosis, NF-|ÊB signaling pathway and glucose metabolism pathways.