异荭草苷通过MAPK/FoxO信号通路减轻LPS诱导的RAW 264.7细胞炎症反应  

Isoorientin reduces inflammatory responses in LPS-induced RAW 264.7 macrophages through MAPK/FoxO signaling pathways

作　　者：杨兰珠 李诗怡 孙雨伟 王勇[2] 杨靖亚[1] YANG Lanzhu;LI Shiyi;SUN Yuwei;WANG Yong;YANG Jingya(College of Food Science and Technology,Shanghai Ocean University,Shanghai 201306;Shanghai Institutes for Biological Sciences,Chinese Academy of Sciences,Shanghai 200032)

机构地区：[1]上海海洋大学食品学院,上海201306 [2]中国科学院上海生命科学研究院,上海200032

出　　处：《安徽农业大学学报》2024年第1期132-137,共6页Journal of Anhui Agricultural University

基　　金：国家重点研发计划(2018YFA0900600)资助。

摘　　要：为了研究异荭草苷(Isoorientin,ISO)对脂多糖(lipopolysaccharide,LPS)诱导的RAW264.7细胞炎症反应的影响及其机制,将对数生长期的RAW264.7细胞随机分为对照组、模型组(LPS 1μg·mL^(-1))和异荭草苷(ISO 2.5~40μg·mL^(-1)+LPS)给药组。各组细胞培养24 h后,MTT法检测ISO对RAW 264.7细胞毒性,Griess法检测细胞培养液中一氧化氮(NO)的含量,ELISA法检测细胞培养液中TNF-α的含量,q RT-PCR方法检测环氧合酶2(COX-2)m RNA的表达水平,蛋白质印迹分析法测定细胞中COX-2、JNK、p-JNK、p38、p-p38、FoxO1和FoxO3蛋白的表达。结果表明,与LPS模型组比较,ISO显著抑制LPS诱导的TNF-α(P<0.001)和NO(P<0.01)的产生,且未见其细胞毒性。此外,ISO以剂量依赖的方式显著抑制RAW 264.7细胞COX-2基因(P<0.001)和蛋白(P<0.05)的表达。蛋白质印迹分析显示,ISO显著抑制MAPK信号通路中p-JNK和p-p38的磷酸化,并减少Fox O1/3的核转位。综上所述,ISO可抑制LPS诱导的RAW 264.7细胞炎症反应,其作用机制可能与其在MAPK/FoxO信号通路中发挥调节作用有关。In order to investigate the effect and mechanism of isoorientin(ISO)on the inflammatory response of RAW 264.7 cells induced by lipopolysaccharide(LPS),RAW264.7 cells in the logarithmic growth phase were randomly divided into a control group,a model group(LPS 1μg·mL^(-1)),and an isoorientin administration groups(ISO 2.5-40μg·mL^(-1)+LPS).After 24 h of cell culture in each group,the cytotoxicity of ISO in RAW 264.7 cells was evaluated by MTT method,the content of nitric oxide(NO)in cell culture medium was detected by Griess method,the supernatant content of tumor necrosis factor-alpha(TNF-α)was detected by ELISA,the mRNA expres-sion levels of COX-2 was detected by qRT-PCR,and the protein expressions of COX-2,JNK,p-JNK,p38,p-p38,FoxO1,and FoxO3 were detected by Western blotting.The results showed that,compared with the LPS model group,ISO significantly inhibited LPS-induced production of TNF-α(P<0.001)and NO(P<0.01),and no cytotoxicity was found.In addition,the expression levels of COX-2 mRNA(P<0.001)and protein(P<0.05)were significantly inhib-ited dose-dependently by ISO in LPS-stimulated RAW 264.7 cells.The Western blotting results showed that ISO significantly attenuated LPS-induced phosphorylation of JNK/p38 MAPK signaling pathway,and reduced the nucle-ar translocation of FoxO1/3.In conclusion,ISO can inhibit LPS-induced inflammatory reaction in RAW 264.7 cells,and the mechanism may be related to its regulatory role in MAPK/FoxO signaling pathway.

关 键 词：异荭草苷 炎症 RAW264.7巨噬细胞 MAPK信号通路 FoxO信号通路 

分 类 号：R285[医药卫生—中药学]