SARS-CoV-2 spike S1 protein promotes IL-6 and IL-8 release via ATP/P2Y2 and ERK1/2 signaling pathways in human bronchial epithelial cells
LIU Xing-jian
FANG Xiao-min
GUO Yu-ling
ZHANG Rui-gang
Abstract：Objective To explore the mechanisms by which SARS-CoV-2 S1 protein induce pro-inflammatory responses in human bronchial epithelial cells and purinegic receptors were involved in the inflammatory by the S1 protein stimulation.Methods The pro-inflammatory cytokine(interleukin-6)IL-6 and(interleukin-8)IL-8 secretion and adenosine triphosphate(ATP)level in16HBE14o-cells induced by S1 protein and/or monoclonal antibody(MAb)were determined by using Enzyme-linked immunosorbent assay(ELISA).The(Heme Oxygenase-1)HO-1,(dual specificity phosphatases-1)MKP-1 and P2Y2 mRNA expression in 16HBE14o-cells after S1 protein stimulation were determined by Real-time quantitative fluorescence PCR(qRT-PCR).Small interfering RNA(siRNA)knockdown P2Y2 expression ELISA detected the IL-6 and IL-8 release in 16HBE14o-cells after S1 protein stimulation.Results S1 protein and MAb hadno significant effects on cell viability(P>0.05).S1 protein induced IL-6 and IL-8 secretion,regulated HO-1,MKP-1,and P2Y2 mRNA expression,and increased extracellular ATP levels in 16HBE14o-cells(P<0.001 or 0.01).The MAb significantly inhibited the secretion of IL-6 and IL-8 induced byS1 protein(P<0.001 or 0.01).siRNA knockdown of P2Y2 expression significantly reduced S1 protein-induced IL-6 and IL-8 secretion,while ERK1/2 inhibitor PD98059 also inhibited S1 protein-induced IL-6 and IL-8 secretion(P<0.001).Conclusion SARS-CoV-2 S1 protein induce inflammatory responses in respiratory epithelial cells through the ATP/P2Y2 and ERK signaling pathways.
Keywords：SARS-CoV-2 S1 proteinextracellular ATPERK1/2bronchial epithelial cellsinflammation
Publication Date：2024-08-28
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：9( 337-345 )