毛蕊花苷对脂多糖诱导RAW264.7细胞炎症模型的作用及其机制研究  

Anti-inflammatory effect and mechanism of verbascoside on LPS-induced RAW264.7 cells

作　　者：王琴琴 韩珊 李新星 高红伟 杨世林 WANG Qin-qin;HAN Shan;LI Xin-xing;GAO Hong-wei;YANG Shi-lin(Faculty of Pharmacy,Guangxi University of Chinese Medicine,Nanning 530020,China;Guangxi Superior Chinese Patent Medicine and National Medicine Development Engineering Technology Center,Nanning 530020,China)

机构地区：[1]广西中医药大学药学院,广西南宁530020 [2]广西优势中成药与民族药开发工程技术中心,广西南宁530020

出　　处：《中草药》2020年第16期4217-4222,4250,共7页Chinese Traditional and Herbal Drugs

基　　金：国家青年科学自然基金项目(81803807);广西自治区青年科学基金项目(2018JJB140265);广西高校引进海外高层次人才“百人计划”项目(05018064);广西科技基地和人才专项(2018AD19034)。

摘　　要：目的采用脂多糖(LPS)诱导RAW264.7细胞的炎症模型,考察炎症相关指标,探讨毛蕊花苷的抗炎作用及其机制。方法用MTT比色法检测毛蕊花苷对RAW264.7细胞活性的影响;Griess法检测NO的含量;Westernblotting检测核转录因子-κB(NF-κB)通路相关蛋白的表达,ELISA试剂盒检测肿瘤坏死因子-α(TNF-α)、白细胞介素-6(IL-6)的释放,流式细胞仪检测细胞内活性氧(ROS)、一氧化氮(NO)、Ca^(2+)的释放情况。结果毛蕊花苷浓度为20、40、80μmol/L时对RAW264.7细胞没有毒性。不同浓度的毛蕊花苷对LPS诱导RAW264.7细胞导致的NO、TNF-α、IL-6的释放,细胞内ROS、NO、Ca^(2+)释放的增加均有很好的抑制效果,对诱导型一氧化氮合酶(i NOS)、环氧化酶-2(COX-2)、p-IKKα/β、p-p65、p-IκBα蛋白的表达也有抑制作用。结论毛蕊花苷具有明显的抗炎作用,其作用机制是通过抑制NF-κB信号通路而发挥抗炎作用。Objective To investigate the anti-inflammatory effects and mechanism of verbascoside in LPS-stimulated RAW264.7 cells. Methods MTT assay was used to determine cytotoxicity of verbascoside in RAW264.7 cells. Griess assay was employed to detect nitrite level in LPS-stimulated RAW264.7 cells. The protein expressions of iNOS, COX-2, p-IKKα/β, p-p65, and p-IκBα were detected by Western blotting. The release of TNF-α and IL-6 was detected by ELISA. The levels of intracellular ROS, NO, and Ca2+ were detected by a flow cytometry. Results Mullein glycoside(20, 40, and 80 μmol/L) displayed no significant cytotoxicity in RAW264.7 cells. The nitrite level, TNF-α and IL-6 release, and intracellular ROS, NO, and Ca2+ level were significantly decreased by Mullein glycoside(20, 40, and 80 μmol/L). In addition, verbascoside also inhibited the protein expressions of iNOS, COX-2, p-IKKα/β, p-p65, and p-IκBα. Conclusion Verbascoside showed an obvious anti-inflammatory effect via the NF-κB signaling pathway.

关 键 词：毛蕊花苷 RAW264.7细胞 脂多糖 抗炎作用 NF-ΚB通路 

分 类 号：R285.5[医药卫生—中药学]