Tilianin downregulated TLR4/Myd88/NF-κB signaling pathway to inhibit NLRP3 inflammasome and inflammatory response
10.16438/j.0513-4870.2024-0228
- VernacularTitle:田蓟苷下调TLR4/Myd88/NF-κB信号通路抑制NLRP3炎症小体和炎症反应
- Author:
Xing-yu ZHANG
1
;
Lei XU
2
;
Kaderyea KADER
2
;
Shou-bao WANG
3
;
Jian-guo XING
2
;
Rui-fang ZHENG
4
Author Information
1. College of Pharmacy, Xinjiang Medical University, Urumqi 830011, China
2. Xinjiang Institute of Pharmacy, Urumqi 830002, China
3. Institute of Materia Medica, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing 100050, China
4. Xinjiang Institute of Pharmacy, Urumqi 830002, China; School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing 211198, China
- Publication Type:Research Article
- Keywords:
tilianin;
lipopolysaccharide;
RAW264.7 cell;
TLR4/Myd88/NF-κB pathway;
NLRP3 inflammasome
- From:
Acta Pharmaceutica Sinica
2024;59(7):2012-2019
- CountryChina
- Language:Chinese
-
Abstract:
In this study, we investigated the anti-inflammatory effect and mechanism of tilianin in lipopolysaccharide (LPS)-induced RAW264.7 cells. The cell viability was detected by cell counting kit-8 (CCK-8) assay. The content of tumor necrosis factor α (TNF-α) and interleukin-6 (IL-6) were detected by enzyme-linked immuno sorbent assay (ELISA) kits. The content of nitric oxide (NO) was assayed by Griess reagent method. The level of intracellular reactive oxygen species (ROS) was detected by 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) fluorescent probe. The protein levels of Toll like receptor 4 (TLR4), myeloid differentiation primary response gene 88 (Myd88), nuclear factors κB p65 (NF-κB p65), phosphorylated nuclear factor κB p65 (p-NF-κB p65), nuclear factor κB inhibitory protein α (IκBα) and phosphorylation nuclear factor κB inhibitory protein α (p-IκBα) were detected by Western blot. The mRNA and protein levels of NLRP3, pro-IL-1β, pro-IL-18 and pro-caspase-1 were detected by qRT-PCR and Western blot. Immunofluorescence staining was used to detect the nuclear translocation of NF-κB p65. The effect of tilianin on the TLR4/Myd88/NF-κB signaling pathway was further validated by using the TLR4 signaling inhibitor restatorvid (TAK242). The results showed that tilianin significantly reduced the levels of TNF-α, IL-6 and NO in the supernatant of RAW264.7 cells and decreased the content of intracellular ROS. Tilianin reduced the levels of TLR4, Myd88, p-NF-κB p65 and p-IκBα protein and nuclear translocation of NF-κB p65. Tilianin could reduce the protein levels of NLRP3, pro-IL-1β, pro-IL-18 and pro-caspase-1. Tilianin significantly inhibited the mRNA levels of NLRP3 and pro-IL-1β. The above research results indicated that tilianin could significantly alleviate the LPS-induced inflammatory response in RAW264.7 cells and its mechanism might be related to downregulate the TLR4/Myd88/NF-κB signaling pathway to inhibit NLRP3 inflammasome.
