Activation of Adenosine A3 Receptor Alleviates TNF-α-Induced Inflammation through Inhibition of the NF-κB Signaling Pathway in Human Colonic Epithelial Cells

Table 2

Summary of the important results.

Items	Results
Cell viability	No significant effect on cell viability was observed at test concentrations of up to 400 nM (𝑃>0.05).
Location and qualitative expression of A3AR, NF-κB p65, IκB-𝛼, and phosphorylated-IκB-𝛼 in HT-29 cells treated with TNF-𝛼, and/or 2-Cl-IB-MECA	TNF-𝛼-stimulated cells pretreated with 2-Cl-IB-MECA showed no obvious change in A3AR expression. Pretreatment with 2-Cl-IB-MECA and subsequent stimulation with TNF-𝛼 attenuated NF-κB p65 nuclear translocation and suppressed the phosphorylation of IκB-𝛼.
mRNA and protein expression of A3AR in HT-29 cells treated with TNF-𝛼 and/or 2-Cl-IB-MECA	The mRNA and protein expression of A3AR showed no significant change among the NC group, TNF-𝛼-only treated group, and 2-Cl-IB-MECA + TNF-𝛼 groups (𝑃>0.05).
Effects of 2-Cl-IB-MECA on TNF-𝛼-induced NF-κB activation	Pretreatment with 2-Cl-IB-MECA prior to stimulation with TNF-𝛼 attenuated NF-κB p65 nuclear translocation as p65 protein decreased in the nucleus of cells and increased in the cytoplasm, inhibited the degradation of IκB-𝛼, and reduced phosphorylated-IκB-𝛼 level, compared to TNF-𝛼-only-treated groups (𝑃<0.05)
Effects of 2-Cl-IB-MECA on TNF-𝛼-induced IL-8 and IL-1𝛽 expression	2-Cl-IB-MECA significantly decreased TNF-𝛼-stimulated IL-8 and IL-1𝛽 mRNA expression and secretion, compared to the TNF-𝛼-only-treated group (𝑃<0.05)