Effect of Tongqiao Decoction on Pyroptosis of Adenoid Epithelial Cells Based on NLRP3 Inflammatory Pathway
Objective To explore the mechanism of Tongqiao Decoction containing serum in inhibiting pyroptosis of adenoid epithelial cells in children with adenoid hypertrophy.Methods The tissue was derived from children with adenoid hypertrophy who underwent adenoidectomy in the Department of Otorhinolaryngology,the First People's Hospital of Yinchuan from March 2022 to March 2023.The discarded adenoid tissue after adenoidectomy was collected,and the epithelial cells were isolated by tissue block adherence method,cultured and identified.The ultrastructure of adenoid epithelial cells was observed by transmission electron microscope.CCK-8 method was used to screen the optimal serum intervention concentration of Tongqiao Decoction.Human primary adenoid epithelial cells cultured to appropriate concentration were divided into four groups:blank control group,montelukast sodium group,Tongqiao Decoction group and MCC950 group(MCC950,10 μmol·L-1).After 24 hours of treatment with corresponding drug-containing serum,the expression levels of NOD-like receptor protein 3(NLRP3),Caspase-1,IL-1β,IL-18 gene and protein were detected by RT-qPCR and Western blot,respectively.Tunnel staining was used to detect cell apoptosis.ELISA method was applied to detect IL-18,IL-1β,IL-6 and TNF-α contents.Results The primary adenoid epithelial cells of children with AH were irregular,unclear in outline,and had poor refractive activity.High positive rate for immunofluorescence cell identification.Transmission electron microscopy showed obvious signs of pyroptosis in adenoid epithelial cells.The cell survival rate was the highest in the low-dose group(P<0.05).Compared with the blank control group,the Tongqiao Decoction group,montelukast sodium group,and MCC950 group had significant reductions in the gene and protein expression of NLRP3,Caspase-1,IL-1β,and IL-18(P all<0.05).The apoptosis rate was decreased(P all<0.05).The levels of IL-18,IL-1β,IL-6 and TNF-α in the cell supernatant were significantly decreased(P all<0.01).Conclusion Tongqiao Decoction containing serum can down-regulate the expression of inflammatory factors by inhibiting pyroptosis of adenoid epithelial cells,and the mechanism may be related to the regulation of NLRP3 mediated pyroptosis signaling pathway.
万方数据
维普
