Effects of total flavonoids of Dracocephalum moldavica L.on ox-LDL-induced inflammatory response of RAW264.7 macrophages via NF-κB/NLRP3 signaling pathway
10.3969/j.issn.1001-1528.2025.02.009
- VernacularTitle: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
- Author:
Yun-li ZHAO
1
;
Chuan-sheng HUANG
1
;
Xin-hong GUO
1
;
Wen-jiang CAO
1
;
Yong YUAN
1
;
Xin-chun WANG
1
Author Information
1. 石河子大学第一附属医院,新疆石河子 832008
- Publication Type:Journal Article
- Keywords:
total flavonoids of Dracocephalum Moldavica L.;
oxidized low density lipoprotein(ox-LDL);
RAW264.7 macrophage cells;
inflammatory reaction;
NF-κB/NLRP3 signaling pathway
- From:
Chinese Traditional Patent Medicine
2025;47(2):413-420
- CountryChina
- Language:Chinese
-
Abstract:
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 factor κB(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(50 μg/mL ox-LDL),the TFDM group(100 μg/mL TFDM+50 μg/mL ox-LDL),the NF-κB inhibitor group(10 μmol/L Bay11-7821+50 μg/mL ox-LDL)and the TFDM+NF-κB inhibitor group(100 μg/mL TFDM+10 μmol/L Bay11-7821+50 μg/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-1β detected by RT-qPCR;their protein expressions of NF-κB p65,IκBα,NLRP3,pro-Caspase-1,Caspase-1,IL-18 and IL-1β by 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 IκBα and cytoplasmic NF-κB p65(P<0.01);increased protein expressions of nuclear NF-κB p65,NLRP3,Caspase-1,IL-1 β and 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 IκBα and cytoplasmic NF-κB p65(P<0.05,P<0.01),decreased protein expressions of nuclear NF-κB p65,NLRP3,Caspase-1,IL-1β and 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 demonstrated decreased mRNA expressions of IL-1βand IL-18(P<0.05),increased IκBα protein expression(P<0.05),decreased protein expressions of nuclear NF-κB p65,NLRP3,Caspase-1,IL-1 β and IL-18(P<0.05),and decreased fluorescence intensity of NLRP3 protein(P<0.05).CONCLUSION TFDM can inhibit the ox-LDL-induced inflammatory response of RAW264.7 macrophages,and the mechansism may be associated with the reduced ROS expression and inflammatory factors due to the inhibited activation of the NF-κB/NLRP3 signaling pathway.