Modified Weijingtang Regulates Pyroptosis of Macrophages via Caspase-1/GSDMD Pathway
10.13422/j.cnki.syfjx.20232422
- VernacularTitle:基于Caspase-1/GSDMD通路探讨加味苇茎汤对巨噬细胞焦亡模型干预作用
- Author:
Dongfang SHANG
1
;
Chenlu ZHAO
2
;
Siying WANG
2
;
Cheng ZHOU
1
;
Minghao LIU
2
;
Pingsheng ZHU
1
;
Suping MA
2
;
Wenxia ZHAO
1
Author Information
1. Henan University of Chinese Medicine， Zhengzhou 450000， China
2. The First Affiliated Hospital of Henan University of Chinese Medicine， Zhengzhou 450000， China
- Publication Type:Journal Article
- Keywords:
modified Weijingtang;
non-alcoholic steatohepatitis;
macrophages;
pyroptosis;
anti-inflammatory effect
- From:
Chinese Journal of Experimental Traditional Medical Formulae
2024;30(11):27-33
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the effect of modified Weijingtang on the pyroptosis of RAW264.7 macrophages via the cysteinyl aspartate-specific protease-1 （Caspase-1）/gasdermin D （GSDMD） pathway. MethodLipopolysaccharide （LPS） was used to induce pyroptosis of RAW264.7 cells. The blank group was treated with the blank serum， and the intervention groups were treated with the sera containing different doses of modified Weijingtang. After 24 h， the viability of cells in different groups was examined by the cell counting kit-8 （CCK-8）. The pyroptosis and morphology of cells in each group were observed by a scanning electron microscope and a phase-contrast microscope， respectively. The mRNA and protein levels of nucleotide-binding oligomerization domain-like receptor protein 3 （NLRP3）， Caspase-1， and GSDMD in each group were determined by real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. The levels of interleukin （IL）-18 and IL-1β in each group were measured by enzyme-linked immunosorbent assay. ResultUnder the electron microscope， RAW264.7 cells presented the best morphology and structure in the blank group and obvious pyroptosis and leakage of cell contents in the model （LPS） group. Compared with the model group， the intervention groups showed reduced pyroptosis to varying degrees， and the high-dose group had the closest cell morphology and structure to the blank group. Under the optical microscope， RAW264.7 cells were spherical in the blank group and irregular with protrusions in the model group. Compared with the model group， the intervention groups showed improved cell morphology， and the cell morphology in the group with the dose of 20% was the closest to that in the blank group. The mRNA and protein levels of NLRP3， Caspase-1， and GSDMD in the model group were higher than those in the blank group （P<0.05）. Compared with the model group， each intervention group showed down-regulated expression of the above indicators （P<0.05）. Compared with the blank group， the model group presented elevated levels of IL-18 and IL-1β （P<0.05）， which were lowered in the intervention （10%， 20%） groups （P<0.01）. ConclusionModified Weijingtang inhibits the pyroptosis of macrophages by down-regulating the Caspase-1/GSDMD pathway and reducing the release of proinflammatory cytokines.
