基于NF-κB/NLRP3信号通路探讨香青兰总黄酮对ox-LDL诱导RAW264.7巨噬细胞炎症反应的影响  

Effects of total flavonoids of Dracocephalum moldavica L.on ox-LDL-induced inflammatory response of RAW264.7 macrophages via NF-κB/NLRP3 signaling pathway

作　　者：赵云丽 黄川生[1] 郭新红[1] 曹文疆[1] 袁勇[1] 王新春[1] ZHAO Yun-li;HUANG Chuan-sheng;GUO Xin-hong;CAO Wen-jiang;YUAN Yong;WANG Xin-chun(The First Hospital Affiliated of Shihezi University,Shihezi 832008,China)

机构地区：[1]石河子大学第一附属医院,新疆石河子832008

出　　处：《中成药》2025年第2期413-420,共8页Chinese Traditional Patent Medicine

基　　金：国家自然科学基金(81960766);兵团重点领域科技攻关计划(2022AB020)。

摘　　要：目的研究香青兰总黄酮(TFDM)通过调节核因子κB(NF-κB)/NOD样受体家族3(NLRP3)信号通路减轻ox-LDL诱导RAW264.7巨噬细胞炎症反应的作用。方法体外培养RAW264.7巨噬细胞,将其分为正常组、模型组(50μg/mL ox-LDL)、香青兰总黄酮组(100μg/mL TFDM+50μg/mL ox-LDL)、NF-κB抑制剂组(10μmol/L Bay11-7821+50μg/mL ox-LDL)、香青兰总黄酮+抑制剂组(100μg/mL TFDM+10μmol/L Bay11-7821+50μg/mL ox-LDL)。采用CCK-8法检测细胞活力,试剂盒测定ROS表达,RT-qPCR法检测细胞NF-κB p65、NLRP3、Caspase-1、IL-18和IL-1βmRNA表达,Western blot法检测细胞NF-κB p65、IκBα、NLRP3、pro-Caspase-1、Caspase-1、IL-18和IL-1β蛋白表达,免疫荧光法检测细胞NF-κB p65和NLRP3蛋白表达。结果与正常组比较,模型组细胞ROS表达升高(P<0.01),NF-κB p65、NLRP3、Caspase-1、IL-18和IL-1βmRNA表达升高(P<0.05,P<0.01),IκBα、胞浆NF-κB p65蛋白表达降低(P<0.01),细胞核NF-κB p65、NLRP3、Caspase-1、IL-1β和IL-18蛋白表达升高(P<0.01),NF-κB p65和NLRP3蛋白荧光强度增强(P<0.01)。与模型组比较,香青兰总黄酮组和香青兰总黄酮+抑制剂组ROS表达降低(P<0.01);香青兰总黄酮组、NF-κB抑制剂组和香青兰总黄酮+抑制剂组NF-κB p65、NLRP3、Caspase-1、IL-18和IL-1βmRNA表达降低(P<0.05,P<0.01),IκBα、胞浆NF-κB p65蛋白表达升高(P<0.05,P<0.01),细胞核NF-κB p65、NLRP3、Caspase-1、IL-1β和IL-18蛋白表达降低(P<0.05,P<0.01),NF-κB p65和NLRP3蛋白荧光强度减弱(P<0.01)。与香青兰总黄酮组比较,NF-κB抑制剂组各指标无明显变化(P>0.05),香青兰总黄酮+抑制剂组IL-1β和IL-18 mRNA表达降低(P<0.05),IκBα蛋白表达升高(P<0.05),细胞核NF-κB p65、NLRP3、Caspase-1、IL-1β和IL-18蛋白表达降低(P<0.05),NLRP3蛋白免疫荧光强度减弱(P<0.05)。结论香青兰总黄酮可抑制ox-LDL诱导的RAW264.7巨噬细胞炎症反应,该作用可能与减少ROS和炎症因子的表达,抑制NF-κB/NLRP3信号通路的活化�AIM To study the effects of total flavonoids of Dracocephalum Moldavica L.(TFDM)on reducing the inflammatory response of RAW264.7 macrophages induced by ox-LDL via the nuclear factorκB(NF-κB)/NOD-like receptor 3(NLRP3)signaling pathway.METHODS The RAW264.7 macrophages cultured in vitro were divided into the normal group,the model group(50μg/mL ox-LDL),the TFDM group(100μg/mL TFDM+50μg/mL ox-LDL),the NF-κB inhibitor group(10μmol/L Bay11-7821+50μg/mL ox-LDL)and the TFDM+NF-κB inhibitor group(100μg/mL TFDM+10μmol/L Bay11-7821+50μg/mL ox-LDL).The cells had their viability assessed by CCK-8 method;their ROS expression detected by the ROS kit;their mRNA expressions of NF-κB p65,NLRP3,Caspase-1,IL-18 and IL-1βdetected by RT-qPCR;their protein expressions of NF-κB p65,IκBα,NLRP3,pro-Caspase-1,Caspase-1,IL-18 and IL-1βby Western blot;their protein expressions of NF-κB p65 and NLRP3 detected using immunofluorescence method.RESULTS Compared with the normal group,the model group showed increased ROS expression(P<0.01);increased mRNA expressions of NF-κB p65,NLRP3,Caspase-1,IL-18 and IL-1β(P<0.05,P<0.01);decreased protein expressions of IκBαand cytoplasmic NF-κB p65(P<0.01);increased protein expressions of nuclear NF-κB p65,NLRP3,Caspase-1,IL-1βand IL-18(P<0.01);and increased fluorescence intensity of NF-κB p65 and NLRP3(P<0.01).Compared with the model group,the groups intervened with either TFDM or TFDM+inhibitor displayed decreased ROS expression(P<0.01);the groups administrated with TFDM or NF-κB inhibitor,or TFDM+inhibitor showed decreased mRNA expressions of NF-κB p65,NLRP3,Caspase-1,IL-18 and IL-1β(P<0.05,P<0.01),increased protein expressions of IκBαand cytoplasmic NF-κB p65(P<0.05,P<0.01),decreased protein expressions of nuclear NF-κB p65,NLRP3,Caspase-1,IL-1βand IL-18(P<0.05,P<0.01),and decreased fluorescence intensity of NF-κB p65 and NLRP3(P<0.01).There existed no significant group difference between the TFDM group and the NF-κB inhibitor group(P>0.05).The TFDM+inhibitor group de

关 键 词：香青兰总黄酮 ox-LDL RAW264.7巨噬细胞 炎症反应 NF-κB/NLRP3信号通路 

分 类 号：R285.5[医药卫生—中药学]