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；于采用Real-Time PCR,Western Blot测FoxO1转录因子在mR-NA水平以及蛋白水平的含量；③利用脂质体2000将FoxO1 siRNA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2000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2190、1124 ng/mL,干预时间为36 h时分别为2190、1995 ng/mL。此条件下炎症因子含量最高；于Real-Time PCR实验和Western blot实验结果显示,在LPS组FoxO1转录因子mRNA水平是对照组的0.36倍,蛋白水平的表达量是对照组的0.58倍；③Foxo1 siRNA干扰实验结果显示,与对照组相比,干扰组巨噬细胞分泌炎症因子TNF-α含量从782 ng/mL上升到1290 ng/mL,IL-6的含量从161 ng/mL上升到251 ng/mL；4.脂质体转染过表达FoxO1质粒实验显示,与LPS组相比,过表达组巨噬细胞分泌炎症因子TNF-α的含量从1785 ng/mL下降到1329 ng/mL,IL-6含量从486 ng/mL下降到317 ng/mL。结论 LPS刺激巨噬细胞Ana-1产生炎症反应,导致炎症因子的释放,同时下调Foxo1转录因子在mRNA水平和蛋白水平的含量,特异性FoxO1基因沉默可上调炎症因子的释放。过表达FoxO1可以使炎症反应减轻,证明Foxo1转录因子参与巨噬细胞引起的炎症反应。%Objective Through study the content changes of FoxO1 transcription factor in the level of mRNA and pro-tein 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 in-flammation. Methods①Used LPS stimulate the Macrophage(Ana-1 cell) to product model of inflammation ,then detect-ed the inflammatory factor with ELISA. ②Used Real-Time PCR、Western blotting to detect the content of FoxO1 tran-scription factor in level of mRNA and protein. ③Used Lipo2 000 put FoxO1 siRNA in Ana-1 cell Media ,We utilized ELISA to detect the concentration of the inflammation factor IL-6,TNF-α in control group and interference group.4. Used Lipo2 000 put FoxO1 over-expression plasmid in Ana-1 cell, We utilized ELISA to detect the concentration of the inflammation factor IL-6,TNF-α in LPS group and over-expression group. Results ①Elisa test showed: when the concentration of LPS was 50 ng/mL,the content of inflammatory factor TNF-α、IL-6 is factor IL-6、TNF-α is 2 190、1 995 ng/mL. In this condition, the content of inflammatory factor was the maximum;②Real-Time PCR and Western blot test showed: The content of FoxO1 transcription factor in level of mRNA in the interference group was 0.36 times as much as the control group ,the content of FoxO1 transcription factor in level of protein was 0.58 times as much as the control group; ③Foxo1 siRNA interference test showed: compared with the control group, the interference group Ana-1cell secreted the content of inflammatory factors TNF-α was increased from 782 ng/mL to 1 290 ng/mL、IL-6 was in-creased from 161 ng/mL to 251 ng/mL; ④Using Lipo 2 000 transfect the over-expreesion plasmid test showed: Com-pared with LPS group,the over-expression group Ana-1 cell secreted the content of inflammatory factors TNF-αwas dropped from 1 785 ng/mL to 1 329 ng/mL,IL-6 was dropped from 486 ng/mL to 317 ng/mL. Conclusion LPS stimulate the Ana-1 cell ,then cause the inflammatory reaction, inducing the release of inflammation factor, meanwhile down-reg-ulate the Foxo1 transcription factor in level of mRNA.Specific gene sliencing can up-regulate the release of inflammato-ry factors.Foxo1 transcription factor indeed participate in the progress of inflammatory reaction.
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