低氧对脂多糖诱导的小鼠炎性巨噬细胞极化的影响及机制研究  

Effect and its mechanism research of hypoxia on mouse inflammatory macrophage polarization induced by lipopolysaccharide

作　　者：谢咏梅[1] 葛化冰[1] 冯㛃 刘琪 XIE Yong-mei;GE Hua-bing;FENG Jie;LIU Qi(Beijing Stomatological Hospital,Capital Medical University,Beijing 100006,China)

机构地区：[1]首都医科大学附属北京口腔医院,北京100006

出　　处：《临床口腔医学杂志》2022年第3期131-135,共5页Journal of Clinical Stomatology

基　　金：首都医科大学附属北京口腔医院学科建设基金(编号:18-09-13)。

摘　　要：目的:观察低氧对脂多糖诱导的小鼠炎性巨噬细胞表型转化的影响及机制研究.方法:通过磁珠分选方法培养原代小鼠腹腔巨噬细胞,利用脂多糖诱导巨噬细胞的经典方法模拟炎症环境,给予原代小鼠腹腔巨噬细胞和巨噬细胞系RAW264.7常氧(21%O_(2))和低氧(1.5%O_(2))培养,利用Real-Time PCR、Western Blot、免疫组化染色、ELISA等方法观察低氧在炎症环境下对巨噬细胞表型转化的影响,并进行进一步的机制研究.结果:与单纯LPS组相比,低氧可升高LPS诱导的巨噬细胞炎性表型(即M1型巨噬细胞)标志物iNOS的蛋白水平,升高iNOS、TNF-α的mRNA水平,增加炎症因子TNF-α的表达,上调iNOS阳性细胞比例;相关机制上,低氧可以增加线粒体ROS、p-IκK、p-IκB、p-P65、p65的表达.结论:低氧通过线粒体ROS-NF-κB信号通路促进LPS诱导M1型巨噬细胞.Objective:To observe the effect and its mechanism research of hypoxia on mouse inflammatory macrophage polarization induced by lipopolysaccharides(LPS).Methods:To culture the primary mouse peritoneal macrophages by magnetic bead sorting,and to use the classical method of inducing macrophages by LPS to simulate the inflammatory environment,giving primary mouse peritoneal macrophages and macrophage cell line RAW264.7 normal oxygen(21%O_(2))and hypoxia(1.5%O_(2))culture.By using Real-Time PCR,Western Blot,immunohistochemical staining,ELISA and other methods etc.to observe the impact of hypoxia on the phenotypic transformation of macrophages in an inflammatory environment,and also to conduct further mechanism research.Results:Compared with the LPS-only group,hypoxia can increase the protein level of the macrophage inflammatory phenotypic(ie,M1 type macrophages)marker iNOS,which induced by LPS,and also increase the mRNA levels of iNOS and TNF-α,increase the expression of inflammatory factors TNF-α,and up-regulate the proportion of iNOS-positive cells.In related mechanisms,hypoxia can increase the expression of mitochondrial ROS,p-IκK,p-IκB,p-P65 and p65.Conclusion:Hypoxia promotes the LPS to induce M1 macrophages through the mitochondrial ROS-NF-κB signaling pathway.

关 键 词：低氧 M1型巨噬细胞 活性氧 线粒体 NF-ΚB 

分 类 号：Q813.1[生物学—生物工程]