基于^1H NMR代谢组学技术探究GRP78蛋白对巨噬细胞极化的干预效应  

Investigation of the Intervention of GRP78 on Macrophage using ~1H NMR-based Metabolomics

作　　者：张立超[1] 李爱平[2] 剌晓琴 李卓玉[1] 

机构地区：[1]山西大学生物技术研究所,化学生物学与分子工程教育部重点实验室,太原030006 [2]山西大学中医药现代研究中心重点实验室,山西太原030006

出　　处：《山西大学学报（自然科学版）》2017年第1期160-168,共9页Journal of Shanxi University(Natural Science Edition)

基　　金：山西省平台项目(2015091015);山西省留学基金(2015-7)

摘　　要：巨噬细胞是肿瘤组织中一类十分重要的免疫细胞,它的M2型极化与肿瘤的恶化密切相关。GRP78蛋白作为肿瘤晚期的生物标志物,可以通过影响肿瘤微环境来促进肿瘤恶化,但GRP78对肿瘤组织中巨噬细胞的分化是否具有干预作用依然未知。本研究利用体外纯化的GRP78蛋白来模拟肿瘤细胞分泌的GRP78,用其处理鼠源巨噬细胞RAW264.7,然后通过~1 H NMR技术的代谢组学方法发现经GRP78处理后细胞中有18种代谢产物,培养基中有14种代谢产物的含量发生了明显变化。通过进一步的代谢通路分析,发现在GRP78蛋白处理后,RAW264.7细胞的糖酵解代谢明显减弱,而脂肪酸和氨基酸代谢加强,这与M2型巨噬细胞的代谢类型一致,表明经GRP78处理后,RAW264.7巨噬细胞向M2型分化。这一发现表明肿瘤微环境中GRP78的存在是肿瘤相关巨噬细胞以M2型为主的重要原因,这也为靶向GRP78的肿瘤治疗提供了坚实的理论依据。Macrophage is one of important immunocytes in tumor,and the M2 macrophage is closely related with tumor progression.GRP78,as a biomarker of advanced tumor,can switch tumor microenvironment to facilitate tumor progression,however,whether GRP78 can affect the macrophage polarization is still unknown.The recombinant GRP78 that mimics the tumor cell secreted-GRP78 was applied to incubate with RAW 264.7cell,then we found 18 metabolites in cell and 14 metabolites in medium,which were different with the control using ^1 H NMR-based metabolomics.Through further analysis of metabolic pathway,we found the glycolysis decreased significantly in RAW264.7with GRP78 treatment while fatty acid and amino acid metabolism increased,which was in accordance with the metabolic type of M2 macrophage,implying that the RAW264.7polarized into M2 type after GRP78 treatment.This study suggested that the tumor cell-secreted GRP78 may be primarily responsible for the M2 polarization of tumor-associated macrophages,and it also provides a solid theoretical basis for GRP78-targeted oncotherapy.

关 键 词：^1H NMR 代谢组学 GRP78 巨噬细胞 分化 

分 类 号：Q7[生物学—分子生物学]