"Trichosanthis Fructus-Allii Macrostemonis Bulbus" combination inhibits NLRP3 inflammasome activation and reduces inflammatory response in RAW264.7 macrophage by inducing autophagy.
10.19540/j.cnki.cjcmm.20230103.707
- Author:
You-Li BAO
1
;
Yin CAO
1
;
Hong-Fei WU
1
Author Information
1. Anhui Province Key Laboratory of Research & Development of Chinese Medicine, Department of Pharmacy, Anhui University of Chinese Medicine Hefei 230012, China.
- Publication Type:Journal Article
- Keywords:
NLRP3 inflammasome;
Trichosanthis Fructus-Allii Macrostemonis;
atherosclerosis;
autophagy;
macrophage inflammation
- MeSH:
Inflammasomes/metabolism*;
NLR Family, Pyrin Domain-Containing 3 Protein/metabolism*;
Interleukin-18/metabolism*;
Lipopolysaccharides/pharmacology*;
Macrophages;
Cytokines/metabolism*;
Caspase 1/metabolism*;
Autophagy;
Interleukin-1beta/metabolism*
- From:
China Journal of Chinese Materia Medica
2023;48(10):2820-2828
- CountryChina
- Language:Chinese
-
Abstract:
This study aims to explore the effect of "Trichosanthis Fructus-Allii Macrostemonis" combination(GX) on the activation of NOD-, LRR-, and pyrin domain-containing protein 3(NLRP3) inflammasome, the release of inflammatory cytokines, and the level of autophagy in RAW264.7 macrophage damaged by lipopolysaccharide(LPS), and the mechanism of GX against inflammatory response in macrophages. To be specific, LPS was used to induce the injury of RAW264.7 cells. Cell Counting Kit-8(CCK-8) assay was employed to measure the survival rate of cells, and Western blot to detect the protein expression of NLRP3, apoptosis-associated speck-like protein(ASC), cysteine-aspartic acid protease(caspase)-1, interleukin(IL)-18, IL-1β, microtubule-associated protein light chain 3(LC3)-Ⅱ, and selective autophagy junction protein p62/sequestosome 1 in RAW264.7 macrophages. ELISA was used to measure the levels of IL-18 and IL-1β in RAW264.7 cells. Transmission electron microscopy was applied to observe the number of autophagosomes in RAW264.7 cells. Immunofulourescence staining was used to detect the expression of LC3-Ⅱ and p62 in RAW264.7 cells. The result showed that GX significantly reduced the protein expression of NLRP3, ASC, and caspase-1 in RAW264.7 cells, significantly increased the protein expression of LC3Ⅱ, decreased the expression of p62, significantly inhibited the secretion of IL-18 and IL-1β, significantly increased the number of autophagosomes, significantly enhanced the immunofluorescence of LC3Ⅱ, and reduced the immunofluorescence of p62. Furthermore, 3-methyladenine(3-MA) could reverse the inhibitory effect of GX on NLRP3, ASC, and caspase-1 and reduce the release of IL-18 and IL-1β. In summary, GX can increase of the autophagy activity of RAW264.7 and inhibit the activation of NLRP3 inflammasome, thereby reducing the release of inflammatory cytokines and suppressing inflammatory response in macrophages.
