目的通过研究叉头框蛋白1(forkhead transcription factors of O class 1,FoxO1)转录因子mRNA水平和蛋白水平在巨噬细胞的表达以及检测Foxo1干扰和过表达后巨噬细胞分泌的炎症因子含量,从炎症进展的角度探讨FoxO1转录因子与炎症之间的关系。方法①利用脂多糖(lipopolysaccharide,LPS)刺激巨噬细胞Ana-1制作炎症模型,ELISA测炎症因子IL-6、TNF-α的浓度;②采用Real-Time PCR,Western Blot测FoxO1转录因子在mRNA水平以及蛋白水平的含量;③利用脂质体2 000将Fox O1 si RNA转染Ana-1细胞,采用ELISA检测对照组和干扰组分泌的炎症因子IL-6、TNF-α的含量;④利用脂质体2 000将FoxO1过表达质粒转染Ana-1细胞,采用ELISA检测LPS组和过表达组炎症因子IL-6、TNF-α的含量。结果①ELISA实验显示,在LPS的浓度为50 ng/mL时巨噬细胞分泌的炎症因子IL-6、TNF-α含量分别为2 190、1 124 ng/mL,干预时间为36 h时分别为2 190、1 995...
【英文摘要】
Objective Through study the content changes of FoxO1 transcription factor in the level of m RNA and protein in Macrophage as well as after the interference of the Foxo1 siRNA detective the content of inflammatory factors.From the perspective of inflammation progress to discuss the relationship between Foxo1 transcription factor and inflammation. Methods①Used LPS stimulate the Macrophage(Ana-1 cell) to product model of inflammation,then detected the inflammatory factor with ELISA.②Used Real-Time PCR、Western ...