牡蒿对脂多糖诱导的RAW 264.7细胞炎症模型的抗炎作用及其机制  

Anti-inflammatory effects and mechanisms of Artemisia japonica on lipopolysaccharideinduced RAW 264.7 macrophages

作　　者：申文静 郭容 龚松 向微 张书环 汪思齐 SHEN Wenjing;GUO Rong;GONG Song;XIANG Wei;ZHANG Shuhuan;WANG Siqi(School of Basic Medicine,Guizhou University of Traditional Chinese Medicine,Guiyang 550055,China;School of Pharmacy,Guizhou University of Traditional Chinese Medicine,Guiyang 550055,China;Judicial Appraisal Center,Guizhou University of Traditional Chinese Medicine,Guiyang 550000,China)

机构地区：[1]贵州中医药大学基础医学院,贵州贵阳550055 [2]贵州中医药大学药学院,贵州贵阳550055 [3]贵州中医药大学司法鉴定所,贵州贵阳550000

出　　处：《现代药物与临床》2025年第1期9-14,共6页Drugs & Clinic

基　　金：国家自然科学基金资助项目(32260925);贵州省科技计划项目(黔科合基础-ZK[2022]一般507);贵州中医药大学大学生创新创业训练计划项目[贵中医大创合字(2021)84号]。

摘　　要：目的基于脂多糖诱导小鼠单核巨噬细胞(RAW 264.7)构建的体外细胞炎症模型,探讨牡蒿Artemisia japonica Thunb.水提物的抗炎作用及其机制。方法通过CCK-8法检测牡蒿水提物对RAW 264.7细胞活力的影响以筛选合适的牡蒿水提物给药质量浓度。采用脂多糖200 ng/mL诱导RAW 264.7细胞建立体外炎症细胞模型。将细胞分为对照组、模型组和牡蒿水提物(240、480、960μg/mL)组,用Griess试剂法检测RAW 264.7细胞一氧化氮(NO)释放水平,ELISA法检测细胞白细胞介素(IL)-6、IL-1β含量。采用Western blotting法检测诱导型一氧化氮合成酶(iNOS)、环氧合酶-2(COX-2)以及Toll样受体4(TLR4)/核因子-κB(NF-κB)信号通路中TLR4、核因子κB抑制蛋白α(IκBα)、p-IκBα、p65蛋白(p65)、p-p65蛋白相对表达量。结果牡蒿水提物质量浓度为0~960μg/mL时对RAW 264.7细胞活力没有显著影响。与模型组相比,牡蒿水提物480、960μg/mL组能显著抑制IL-6的释放以及COX-2的蛋白相对表达量(P<0.01);各质量浓度牡蒿水提物均能显著抑制NO、IL-1β的释放,iNOS、p-IκBα、p65、p-p65蛋白的表达以及IκBα蛋白的降解(P<0.01)。结论牡蒿水提物能抑制脂多糖诱导的RAW 264.7细胞炎性因子的分泌,其作用机制与抑制炎性蛋白iNOS和COX-2的表达及TLR4/NF-κB信号通路的激活有关。Objective To explore the effects and mechanisms of A.japonica water extract on lipopolysaccharide-induced RAW 264.7 macrophages model.Methods CCK-8 colorimetric method was used to select the appropriate concentration of A.japonica on the activity of RAW 264.7 macrophages.RAW 264.7 cells were induced by lipopolysaccharide 200 ng/mL to establish in vitro inflammatory cell model.Cells were divided into control group,model group,and A.japonica(240,480,and 960μg/mL)groups.Griess assay was used to assess NO concentration,IL-1β,and IL-6 content was measured by ELISA.The expression of iNOS,COX2,and TLR4/NF-κB pathway-related proteins(TLR4,IκBα,p-IκBα,p65,p-P65)were detected by Western blotting.Results A.japonica had no significant effect on the viability of RAW 264.7 cells at the concentrations of 0—960μg/mL.Compared with model group,A.japonica water extract 480 and 960μg/mL groups significantly inhibitory effects on the release of IL-6 and protein expression of COX-2(P<0.01).Each dose group of A.japonica water extract displayed favorable inhibitory decreased effects on release of NO and IL-1β,expression of iNOS,p-IκBα,p65,and p-p65 proteins and degradation of IκBα.Conclusion A.japonica water extract could inhibit lipopolysaccharide-induced secretion of inflammatory factors in RAW 264.7 cells,the mechanism of which is related to inhibiting the expression of inflammatory proteins iNOS and COX-2 and inhibiting the activation of TLR4/NF-κB signaling pathway.

关 键 词：牡蒿 脂多糖 RAW 264.7细胞 白细胞介素-6、白细胞介素-1β Toll样受体4/核因子κB信号通路 

分 类 号：R285.5[医药卫生—中药学]